var bibbase_data = {"data":"<img src=\"//bibbase.org/img/ajax-loader.gif\" id=\"spinner\"\n  style=\"display: none;\" alt=\"Loading..\" />\n\n<div id=\"bibbase\">\n\n  <script type=\"text/javascript\">\n    var bibbase = {\n      params: {\"jsonp\":\"1\",\"host\":\"bibbase.org\",\"ssl\":false},\n      data: [{\"type\":\"inbook\",\"year\":\"2010\",\"keywords\":\"RNA-binding domain,Zinc fingers,protein design,protein interaction domain\",\"pages\":\"479-491\",\"volume\":\"649\",\"websites\":\"http://link.springer.com/10.1007/978-1-60761-753-2_29\",\"id\":\"0f1743b7-0299-3ee3-8152-d528a8b42877\",\"created\":\"2020-12-17T05:29:52.309Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:52.309Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Font2010\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 21\",\"private_publication\":false,\"abstract\":\"Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.\",\"bibtype\":\"inbook\",\"author\":\"Font, Josep and Mackay, Joel P.\",\"doi\":\"10.1007/978-1-60761-753-2_29\",\"chapter\":\"Beyond DNA: Zinc Finger Domains as RNA-Binding Modules\",\"title\":\"Methods in Molecular Biology\",\"bibtex\":\"@inbook{\\n type = {inbook},\\n year = {2010},\\n keywords = {RNA-binding domain,Zinc fingers,protein design,protein interaction domain},\\n pages = {479-491},\\n volume = {649},\\n websites = {http://link.springer.com/10.1007/978-1-60761-753-2_29},\\n id = {0f1743b7-0299-3ee3-8152-d528a8b42877},\\n created = {2020-12-17T05:29:52.309Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:52.309Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Font2010},\\n source_type = {ARTICLE},\\n notes = {cited By 21},\\n private_publication = {false},\\n abstract = {Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.},\\n bibtype = {inbook},\\n author = {Font, Josep and Mackay, Joel P.},\\n doi = {10.1007/978-1-60761-753-2_29},\\n chapter = {Beyond DNA: Zinc Finger Domains as RNA-Binding Modules},\\n title = {Methods in Molecular Biology}\\n}\",\"author_short\":[\"Font,  J.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://link.springer.com/10.1007/978-1-60761-753-2_29\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"font-mackay-methodsinmolecularbiology-2010\",\"role\":\"author\",\"keyword\":[\"RNA-binding domain\",\"Zinc fingers\",\"protein design\",\"protein interaction domain\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"1H, 13C and 15N resonance assignments of an N-terminal domain of CHD4\",\"type\":\"article\",\"year\":\"2014\",\"keywords\":\"CHD4,Chromatin remodeling,N-terminal domain,NuRD complex,PAR-binding motif\",\"pages\":\"137-139\",\"volume\":\"8\",\"websites\":\"http://link.springer.com/10.1007/s12104-013-9469-3\",\"month\":\"4\",\"day\":\"17\",\"id\":\"6b604bfa-8cf1-39d7-9959-cdd1ad06e0c1\",\"created\":\"2020-12-17T05:29:52.354Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:52.354Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Silva2014\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 1\",\"private_publication\":false,\"abstract\":\"Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.\",\"bibtype\":\"article\",\"author\":\"Silva, Ana P. G. and Kwan, Ann H. and Mackay, Joel P.\",\"doi\":\"10.1007/s12104-013-9469-3\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {1H, 13C and 15N resonance assignments of an N-terminal domain of CHD4},\\n type = {article},\\n year = {2014},\\n keywords = {CHD4,Chromatin remodeling,N-terminal domain,NuRD complex,PAR-binding motif},\\n pages = {137-139},\\n volume = {8},\\n websites = {http://link.springer.com/10.1007/s12104-013-9469-3},\\n month = {4},\\n day = {17},\\n id = {6b604bfa-8cf1-39d7-9959-cdd1ad06e0c1},\\n created = {2020-12-17T05:29:52.354Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:52.354Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Silva2014},\\n source_type = {ARTICLE},\\n notes = {cited By 1},\\n private_publication = {false},\\n abstract = {Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.},\\n bibtype = {article},\\n author = {Silva, Ana P. G. and Kwan, Ann H. and Mackay, Joel P.},\\n doi = {10.1007/s12104-013-9469-3},\\n journal = {Biomolecular NMR Assignments},\\n number = {1}\\n}\",\"author_short\":[\"Silva,  A., P., G.\",\"Kwan,  A., H.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://link.springer.com/10.1007/s12104-013-9469-3\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"silva-kwan-mackay-1h13cand15nresonanceassignmentsofannterminaldomainofchd4-2014\",\"role\":\"author\",\"keyword\":[\"CHD4\",\"Chromatin remodeling\",\"N-terminal domain\",\"NuRD complex\",\"PAR-binding motif\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The Hydrophobin EAS Is Largely Unstructured in Solution and Functions by Forming Amyloid-Like Structures\",\"type\":\"article\",\"year\":\"2001\",\"keywords\":\"Amphipathic monolayer,EAS,Neurospora crassa,Self-assembly,hydrophobin\",\"pages\":\"83-91\",\"volume\":\"9\",\"websites\":\"https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591\",\"month\":\"2\",\"id\":\"298718cc-0cb5-316f-89d4-dead241698cf\",\"created\":\"2020-12-17T05:29:52.461Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:52.461Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Mackay2001\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 115\",\"private_publication\":false,\"abstract\":\"Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.\",\"bibtype\":\"article\",\"author\":\"Mackay, Joel P and Matthews, Jacqueline M. and Winefield, Robert D. and Mackay, Lindsey G. and Haverkamp, Richard G. and Templeton, Matthew D.\",\"doi\":\"10.1016/S0969-2126(00)00559-1\",\"journal\":\"Structure\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {The Hydrophobin EAS Is Largely Unstructured in Solution and Functions by Forming Amyloid-Like Structures},\\n type = {article},\\n year = {2001},\\n keywords = {Amphipathic monolayer,EAS,Neurospora crassa,Self-assembly,hydrophobin},\\n pages = {83-91},\\n volume = {9},\\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591},\\n month = {2},\\n id = {298718cc-0cb5-316f-89d4-dead241698cf},\\n created = {2020-12-17T05:29:52.461Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:52.461Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Mackay2001},\\n source_type = {ARTICLE},\\n notes = {cited By 115},\\n private_publication = {false},\\n abstract = {Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.},\\n bibtype = {article},\\n author = {Mackay, Joel P and Matthews, Jacqueline M. and Winefield, Robert D. and Mackay, Lindsey G. and Haverkamp, Richard G. and Templeton, Matthew D.},\\n doi = {10.1016/S0969-2126(00)00559-1},\\n journal = {Structure},\\n number = {2}\\n}\",\"author_short\":[\"Mackay,  J., P.\",\"Matthews,  J., M.\",\"Winefield,  R., D.\",\"Mackay,  L., G.\",\"Haverkamp,  R., G.\",\"Templeton,  M., D.\"],\"urls\":{\"Website\":\"https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001\",\"role\":\"author\",\"keyword\":[\"Amphipathic monolayer\",\"EAS\",\"Neurospora crassa\",\"Self-assembly\",\"hydrophobin\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex\",\"type\":\"article\",\"year\":\"2017\",\"keywords\":\"chromatin remodelling,co-immunoprecipitations,nucleosome remodelling and deacetylase (NuRD) comp,protein structure,protein–protein interactions\",\"pages\":\"4216-4232\",\"volume\":\"284\",\"websites\":\"http://doi.wiley.com/10.1111/febs.14301\",\"month\":\"12\",\"id\":\"fa40e576-8ce6-3e7e-83ec-453c57f0abe1\",\"created\":\"2020-12-17T05:29:52.498Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:52.498Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Torrado2017\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 4\",\"private_publication\":false,\"abstract\":\"The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.\",\"bibtype\":\"article\",\"author\":\"Torrado, Mario and Low, Jason K. K. and Silva, Ana P. G. and Schmidberger, Jason W. and Sana, Maryam and Sharifi Tabar, Mehdi and Isilak, Musa E. and Winning, Courtney S. and Kwong, Cherry and Bedward, Max J. and Sperlazza, Mary J. and Williams, David C. and Shepherd, Nicholas E. and Mackay, Joel P.\",\"doi\":\"10.1111/febs.14301\",\"journal\":\"The FEBS Journal\",\"number\":\"24\",\"bibtex\":\"@article{\\n title = {Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex},\\n type = {article},\\n year = {2017},\\n keywords = {chromatin remodelling,co-immunoprecipitations,nucleosome remodelling and deacetylase (NuRD) comp,protein structure,protein–protein interactions},\\n pages = {4216-4232},\\n volume = {284},\\n websites = {http://doi.wiley.com/10.1111/febs.14301},\\n month = {12},\\n id = {fa40e576-8ce6-3e7e-83ec-453c57f0abe1},\\n created = {2020-12-17T05:29:52.498Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:52.498Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Torrado2017},\\n source_type = {ARTICLE},\\n notes = {cited By 4},\\n private_publication = {false},\\n abstract = {The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.},\\n bibtype = {article},\\n author = {Torrado, Mario and Low, Jason K. K. and Silva, Ana P. G. and Schmidberger, Jason W. and Sana, Maryam and Sharifi Tabar, Mehdi and Isilak, Musa E. and Winning, Courtney S. and Kwong, Cherry and Bedward, Max J. and Sperlazza, Mary J. and Williams, David C. and Shepherd, Nicholas E. and Mackay, Joel P.},\\n doi = {10.1111/febs.14301},\\n journal = {The FEBS Journal},\\n number = {24}\\n}\",\"author_short\":[\"Torrado,  M.\",\"Low,  J., K., K.\",\"Silva,  A., P., G.\",\"Schmidberger,  J., W.\",\"Sana,  M.\",\"Sharifi Tabar,  M.\",\"Isilak,  M., E.\",\"Winning,  C., S.\",\"Kwong,  C.\",\"Bedward,  M., J.\",\"Sperlazza,  M., J.\",\"Williams,  D., C.\",\"Shepherd,  N., E.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1111/febs.14301\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017\",\"role\":\"author\",\"keyword\":[\"chromatin remodelling\",\"co-immunoprecipitations\",\"nucleosome remodelling and deacetylase (NuRD) comp\",\"protein structure\",\"protein–protein interactions\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean\",\"type\":\"article\",\"year\":\"2017\",\"keywords\":\"CLE,CLE40,IAD,arabinosylation,glycopeptide,glycosylation,hormone,legume,root apical meristem,soybean\",\"pages\":\"1347-1355.e7\",\"volume\":\"24\",\"websites\":\"https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124\",\"month\":\"11\",\"id\":\"9da321e9-ae64-34bc-8487-6b2f91bcf82c\",\"created\":\"2020-12-17T05:29:52.604Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:52.604Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Corcilius2017\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 8\",\"private_publication\":false,\"abstract\":\"Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.\",\"bibtype\":\"article\",\"author\":\"Corcilius, Leo and Hastwell, April H. and Zhang, Mengbai and Williams, James and Mackay, Joel P. and Gresshoff, Peter M. and Ferguson, Brett J. and Payne, Richard J.\",\"doi\":\"10.1016/j.chembiol.2017.08.014\",\"journal\":\"Cell Chemical Biology\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean},\\n type = {article},\\n year = {2017},\\n keywords = {CLE,CLE40,IAD,arabinosylation,glycopeptide,glycosylation,hormone,legume,root apical meristem,soybean},\\n pages = {1347-1355.e7},\\n volume = {24},\\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124},\\n month = {11},\\n id = {9da321e9-ae64-34bc-8487-6b2f91bcf82c},\\n created = {2020-12-17T05:29:52.604Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:52.604Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Corcilius2017},\\n source_type = {ARTICLE},\\n notes = {cited By 8},\\n private_publication = {false},\\n abstract = {Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.},\\n bibtype = {article},\\n author = {Corcilius, Leo and Hastwell, April H. and Zhang, Mengbai and Williams, James and Mackay, Joel P. and Gresshoff, Peter M. and Ferguson, Brett J. and Payne, Richard J.},\\n doi = {10.1016/j.chembiol.2017.08.014},\\n journal = {Cell Chemical Biology},\\n number = {11}\\n}\",\"author_short\":[\"Corcilius,  L.\",\"Hastwell,  A., H.\",\"Zhang,  M.\",\"Williams,  J.\",\"Mackay,  J., P.\",\"Gresshoff,  P., M.\",\"Ferguson,  B., J.\",\"Payne,  R., J.\"],\"urls\":{\"Website\":\"https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017\",\"role\":\"author\",\"keyword\":[\"CLE\",\"CLE40\",\"IAD\",\"arabinosylation\",\"glycopeptide\",\"glycosylation\",\"hormone\",\"legume\",\"root apical meristem\",\"soybean\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The NuRD complex and macrocephaly associated neurodevelopmental disorders\",\"type\":\"article\",\"year\":\"2019\",\"keywords\":\"CHD3,CHD4,GATAD2B,NuRD complex,macrocephaly\",\"pages\":\"548-556\",\"volume\":\"181\",\"websites\":\"http://doi.org/10.1002/ajmg.c.31752\",\"month\":\"12\",\"day\":\"18\",\"id\":\"786626bb-912b-3ca7-aa02-82a98a6afa89\",\"created\":\"2020-12-17T05:29:52.824Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:52.824Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Pierson2019\",\"source_type\":\"article\",\"private_publication\":false,\"abstract\":\"The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.\",\"bibtype\":\"article\",\"author\":\"Pierson, Tyler Mark and Otero, Maria G. and Grand, Katheryn and Choi, Andrew and Graham, John M. and Young, Juan I. and Mackay, Joel P.\",\"doi\":\"10.1002/ajmg.c.31752\",\"journal\":\"American Journal of Medical Genetics Part C: Seminars in Medical Genetics\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {The NuRD complex and macrocephaly associated neurodevelopmental disorders},\\n type = {article},\\n year = {2019},\\n keywords = {CHD3,CHD4,GATAD2B,NuRD complex,macrocephaly},\\n pages = {548-556},\\n volume = {181},\\n websites = {http://doi.org/10.1002/ajmg.c.31752},\\n month = {12},\\n day = {18},\\n id = {786626bb-912b-3ca7-aa02-82a98a6afa89},\\n created = {2020-12-17T05:29:52.824Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:52.824Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Pierson2019},\\n source_type = {article},\\n private_publication = {false},\\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.},\\n bibtype = {article},\\n author = {Pierson, Tyler Mark and Otero, Maria G. and Grand, Katheryn and Choi, Andrew and Graham, John M. and Young, Juan I. and Mackay, Joel P.},\\n doi = {10.1002/ajmg.c.31752},\\n journal = {American Journal of Medical Genetics Part C: Seminars in Medical Genetics},\\n number = {4}\\n}\",\"author_short\":[\"Pierson,  T., M.\",\"Otero,  M., G.\",\"Grand,  K.\",\"Choi,  A.\",\"Graham,  J., M.\",\"Young,  J., I.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://doi.org/10.1002/ajmg.c.31752\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019\",\"role\":\"author\",\"keyword\":[\"CHD3\",\"CHD4\",\"GATAD2B\",\"NuRD complex\",\"macrocephaly\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Crystallization of a ZRANB2–RNA complex\",\"type\":\"article\",\"year\":\"2008\",\"keywords\":\"RNA-binding proteins,RanBP2-type zinc fingers,Splicing factors\",\"pages\":\"1175-1177\",\"volume\":\"64\",\"websites\":\"http://scripts.iucr.org/cgi-bin/paper?S1744309108036993\",\"month\":\"12\",\"day\":\"1\",\"id\":\"7587f247-d4b3-314f-84e0-30baa59ddf19\",\"created\":\"2020-12-17T05:29:52.864Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:52.864Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Loughlin2008\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 1\",\"private_publication\":false,\"abstract\":\"ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.\",\"bibtype\":\"article\",\"author\":\"Loughlin, Fionna E. and Lee, Mihwa and Guss, J. Mitchell and Mackay, Joel P.\",\"doi\":\"10.1107/S1744309108036993\",\"journal\":\"Acta Crystallographica Section F Structural Biology and Crystallization Communications\",\"number\":\"12\",\"bibtex\":\"@article{\\n title = {Crystallization of a ZRANB2–RNA complex},\\n type = {article},\\n year = {2008},\\n keywords = {RNA-binding proteins,RanBP2-type zinc fingers,Splicing factors},\\n pages = {1175-1177},\\n volume = {64},\\n websites = {http://scripts.iucr.org/cgi-bin/paper?S1744309108036993},\\n month = {12},\\n day = {1},\\n id = {7587f247-d4b3-314f-84e0-30baa59ddf19},\\n created = {2020-12-17T05:29:52.864Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:52.864Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Loughlin2008},\\n source_type = {ARTICLE},\\n notes = {cited By 1},\\n private_publication = {false},\\n abstract = {ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.},\\n bibtype = {article},\\n author = {Loughlin, Fionna E. and Lee, Mihwa and Guss, J. Mitchell and Mackay, Joel P.},\\n doi = {10.1107/S1744309108036993},\\n journal = {Acta Crystallographica Section F Structural Biology and Crystallization Communications},\\n number = {12}\\n}\",\"author_short\":[\"Loughlin,  F., E.\",\"Lee,  M.\",\"Guss,  J., M.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://scripts.iucr.org/cgi-bin/paper?S1744309108036993\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008\",\"role\":\"author\",\"keyword\":[\"RNA-binding proteins\",\"RanBP2-type zinc fingers\",\"Splicing factors\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain\",\"type\":\"article\",\"year\":\"2016\",\"keywords\":\"C17orf64,DNA-binding domain,helical bundle,nucleosomes,transcription\",\"pages\":\"4298-4314\",\"volume\":\"428\",\"websites\":\"https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485\",\"month\":\"10\",\"id\":\"851b1fbc-10b7-3289-b546-77e83ffb0c9d\",\"created\":\"2020-12-17T05:29:53.207Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:53.207Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Mohanty2016\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 5\",\"private_publication\":false,\"abstract\":\"The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.\",\"bibtype\":\"article\",\"author\":\"Mohanty, Biswaranjan and Helder, Stephanie and Silva, Ana P.G. and Mackay, Joel P. and Ryan, Daniel P.\",\"doi\":\"10.1016/j.jmb.2016.08.028\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"21\",\"bibtex\":\"@article{\\n title = {The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain},\\n type = {article},\\n year = {2016},\\n keywords = {C17orf64,DNA-binding domain,helical bundle,nucleosomes,transcription},\\n pages = {4298-4314},\\n volume = {428},\\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485},\\n month = {10},\\n id = {851b1fbc-10b7-3289-b546-77e83ffb0c9d},\\n created = {2020-12-17T05:29:53.207Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:53.207Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Mohanty2016},\\n source_type = {ARTICLE},\\n notes = {cited By 5},\\n private_publication = {false},\\n abstract = {The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.},\\n bibtype = {article},\\n author = {Mohanty, Biswaranjan and Helder, Stephanie and Silva, Ana P.G. and Mackay, Joel P. and Ryan, Daniel P.},\\n doi = {10.1016/j.jmb.2016.08.028},\\n journal = {Journal of Molecular Biology},\\n number = {21}\\n}\",\"author_short\":[\"Mohanty,  B.\",\"Helder,  S.\",\"Silva,  A., P.\",\"Mackay,  J., P.\",\"Ryan,  D., P.\"],\"urls\":{\"Website\":\"https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016\",\"role\":\"author\",\"keyword\":[\"C17orf64\",\"DNA-binding domain\",\"helical bundle\",\"nucleosomes\",\"transcription\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder\",\"type\":\"article\",\"year\":\"2020\",\"keywords\":\"GATAD2B,NuRD complex,apraxia of speech,chromati\",\"pages\":\"878-888\",\"volume\":\"22\",\"websites\":\"http://doi.org/10.1038/s41436-019-0747-z\",\"month\":\"5\",\"day\":\"17\",\"id\":\"6308af4d-95ba-3986-b5f1-b8362afa6d89\",\"created\":\"2020-12-17T05:29:53.237Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:53.237Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Shieh2020\",\"source_type\":\"article\",\"private_publication\":false,\"abstract\":\"Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.\",\"bibtype\":\"article\",\"author\":\"Shieh, Christine and Jones, Natasha and Vanle, Brigitte and Au, Margaret and Huang, Alden Y. and Silva, Ana P. G. and Lee, Hane and Douine, Emilie D. and Otero, Maria G. and Choi, Andrew and Grand, Katheryn and Taff, Ingrid P. and Delgado, Mauricio R. and Hajianpour, M. J. and Seeley, Andrea and Rohena, Luis and Vernon, Hilary and Gripp, Karen W. and Vergano, Samantha A. and Mahida, Sonal and Naidu, Sakkubai and Sousa, Ana Berta and Wain, Karen E. and Challman, Thomas D. and Beek, Geoffrey and Basel, Donald and Ranells, Judith and Smith, Rosemarie and Yusupov, Roman and Freckmann, Mary-Louise and Ohden, Lisa and Davis-Keppen, Laura and Chitayat, David and Dowling, James J. and Finkel, Richard and Dauber, Andrew and Spillmann, Rebecca and Pena, Loren D. M. and Metcalfe, Kay and Splitt, Miranda and Lachlan, Katherine and McKee, Shane A. and Hurst, Jane and Fitzpatrick, David R. and Morton, Jenny E. V. and Cox, Helen and Venkateswaran, Sunita and Young, Juan I. and Marsh, Eric D. and Nelson, Stanley F. and Martinez, Julian A. and Graham, John M. and Kini, Usha and Mackay, Joel P. and Pierson, Tyler Mark\",\"doi\":\"10.1038/s41436-019-0747-z\",\"journal\":\"Genetics in Medicine\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder},\\n type = {article},\\n year = {2020},\\n keywords = {GATAD2B,NuRD complex,apraxia of speech,chromati},\\n pages = {878-888},\\n volume = {22},\\n websites = {http://doi.org/10.1038/s41436-019-0747-z},\\n month = {5},\\n day = {17},\\n id = {6308af4d-95ba-3986-b5f1-b8362afa6d89},\\n created = {2020-12-17T05:29:53.237Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:53.237Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Shieh2020},\\n source_type = {article},\\n private_publication = {false},\\n abstract = {Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.},\\n bibtype = {article},\\n author = {Shieh, Christine and Jones, Natasha and Vanle, Brigitte and Au, Margaret and Huang, Alden Y. and Silva, Ana P. G. and Lee, Hane and Douine, Emilie D. and Otero, Maria G. and Choi, Andrew and Grand, Katheryn and Taff, Ingrid P. and Delgado, Mauricio R. and Hajianpour, M. J. and Seeley, Andrea and Rohena, Luis and Vernon, Hilary and Gripp, Karen W. and Vergano, Samantha A. and Mahida, Sonal and Naidu, Sakkubai and Sousa, Ana Berta and Wain, Karen E. and Challman, Thomas D. and Beek, Geoffrey and Basel, Donald and Ranells, Judith and Smith, Rosemarie and Yusupov, Roman and Freckmann, Mary-Louise and Ohden, Lisa and Davis-Keppen, Laura and Chitayat, David and Dowling, James J. and Finkel, Richard and Dauber, Andrew and Spillmann, Rebecca and Pena, Loren D. M. and Metcalfe, Kay and Splitt, Miranda and Lachlan, Katherine and McKee, Shane A. and Hurst, Jane and Fitzpatrick, David R. and Morton, Jenny E. V. and Cox, Helen and Venkateswaran, Sunita and Young, Juan I. and Marsh, Eric D. and Nelson, Stanley F. and Martinez, Julian A. and Graham, John M. and Kini, Usha and Mackay, Joel P. and Pierson, Tyler Mark},\\n doi = {10.1038/s41436-019-0747-z},\\n journal = {Genetics in Medicine},\\n number = {5}\\n}\",\"author_short\":[\"Shieh,  C.\",\"Jones,  N.\",\"Vanle,  B.\",\"Au,  M.\",\"Huang,  A., Y.\",\"Silva,  A., P., G.\",\"Lee,  H.\",\"Douine,  E., D.\",\"Otero,  M., G.\",\"Choi,  A.\",\"Grand,  K.\",\"Taff,  I., P.\",\"Delgado,  M., R.\",\"Hajianpour,  M., J.\",\"Seeley,  A.\",\"Rohena,  L.\",\"Vernon,  H.\",\"Gripp,  K., W.\",\"Vergano,  S., A.\",\"Mahida,  S.\",\"Naidu,  S.\",\"Sousa,  A., B.\",\"Wain,  K., E.\",\"Challman,  T., D.\",\"Beek,  G.\",\"Basel,  D.\",\"Ranells,  J.\",\"Smith,  R.\",\"Yusupov,  R.\",\"Freckmann,  M.\",\"Ohden,  L.\",\"Davis-Keppen,  L.\",\"Chitayat,  D.\",\"Dowling,  J., J.\",\"Finkel,  R.\",\"Dauber,  A.\",\"Spillmann,  R.\",\"Pena,  L., D., M.\",\"Metcalfe,  K.\",\"Splitt,  M.\",\"Lachlan,  K.\",\"McKee,  S., A.\",\"Hurst,  J.\",\"Fitzpatrick,  D., R.\",\"Morton,  J., E., V.\",\"Cox,  H.\",\"Venkateswaran,  S.\",\"Young,  J., I.\",\"Marsh,  E., D.\",\"Nelson,  S., F.\",\"Martinez,  J., A.\",\"Graham,  J., M.\",\"Kini,  U.\",\"Mackay,  J., P.\",\"Pierson,  T., M.\"],\"urls\":{\"Website\":\"http://doi.org/10.1038/s41436-019-0747-z\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020\",\"role\":\"author\",\"keyword\":[\"GATAD2B\",\"NuRD complex\",\"apraxia of speech\",\"chromati\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P\",\"type\":\"article\",\"year\":\"2018\",\"keywords\":\"NLR [nucleotide-binding and oligomerization domain,crystal structure,effector-triggered immunity,flax rust (Melampsora lini) effector,nuclear localization,nucleotide-binding/leucine-rich repeat receptor],plant disease resistance,zinc finger\",\"pages\":\"1196-1209\",\"volume\":\"19\",\"websites\":\"http://doi.wiley.com/10.1111/mpp.12597\",\"month\":\"5\",\"id\":\"67c03f3e-7a41-34e7-89cf-55716f13ac32\",\"created\":\"2020-12-17T05:29:53.248Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:53.248Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Zhang2018\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 2\",\"private_publication\":false,\"abstract\":\"The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.\",\"bibtype\":\"article\",\"author\":\"Zhang, Xiaoxiao and Farah, Nadya and Rolston, Laura and Ericsson, Daniel J. and Catanzariti, Ann-Maree and Bernoux, Maud and Ve, Thomas and Bendak, Katerina and Chen, Chunhong and Mackay, Joel P. and Lawrence, Gregory J. and Hardham, Adrienne and Ellis, Jeffrey G. and Williams, Simon J. and Dodds, Peter N. and Jones, David A. and Kobe, Bostjan\",\"doi\":\"10.1111/mpp.12597\",\"journal\":\"Molecular Plant Pathology\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P},\\n type = {article},\\n year = {2018},\\n keywords = {NLR [nucleotide-binding and oligomerization domain,crystal structure,effector-triggered immunity,flax rust (Melampsora lini) effector,nuclear localization,nucleotide-binding/leucine-rich repeat receptor],plant disease resistance,zinc finger},\\n pages = {1196-1209},\\n volume = {19},\\n websites = {http://doi.wiley.com/10.1111/mpp.12597},\\n month = {5},\\n id = {67c03f3e-7a41-34e7-89cf-55716f13ac32},\\n created = {2020-12-17T05:29:53.248Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:53.248Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Zhang2018},\\n source_type = {ARTICLE},\\n notes = {cited By 2},\\n private_publication = {false},\\n abstract = {The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.},\\n bibtype = {article},\\n author = {Zhang, Xiaoxiao and Farah, Nadya and Rolston, Laura and Ericsson, Daniel J. and Catanzariti, Ann-Maree and Bernoux, Maud and Ve, Thomas and Bendak, Katerina and Chen, Chunhong and Mackay, Joel P. and Lawrence, Gregory J. and Hardham, Adrienne and Ellis, Jeffrey G. and Williams, Simon J. and Dodds, Peter N. and Jones, David A. and Kobe, Bostjan},\\n doi = {10.1111/mpp.12597},\\n journal = {Molecular Plant Pathology},\\n number = {5}\\n}\",\"author_short\":[\"Zhang,  X.\",\"Farah,  N.\",\"Rolston,  L.\",\"Ericsson,  D., J.\",\"Catanzariti,  A.\",\"Bernoux,  M.\",\"Ve,  T.\",\"Bendak,  K.\",\"Chen,  C.\",\"Mackay,  J., P.\",\"Lawrence,  G., J.\",\"Hardham,  A.\",\"Ellis,  J., G.\",\"Williams,  S., J.\",\"Dodds,  P., N.\",\"Jones,  D., A.\",\"Kobe,  B.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1111/mpp.12597\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018\",\"role\":\"author\",\"keyword\":[\"NLR [nucleotide-binding and oligomerization domain\",\"crystal structure\",\"effector-triggered immunity\",\"flax rust (Melampsora lini) effector\",\"nuclear localization\",\"nucleotide-binding/leucine-rich repeat receptor]\",\"plant disease resistance\",\"zinc finger\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/Friend of GATA interaction\",\"type\":\"article\",\"year\":\"2005\",\"keywords\":\"Factor,Gene expression,Transcription\",\"pages\":\"583-588\",\"volume\":\"102\",\"websites\":\"http://www.ncbi.nlm.nih.gov/pubmed/15644435\",\"month\":\"1\",\"day\":\"18\",\"id\":\"4fa93b4d-9743-397c-8bc8-7dec216c8aa7\",\"created\":\"2020-12-17T05:29:53.577Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:53.577Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Liew2005\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 65\",\"private_publication\":false,\"abstract\":\"GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.\",\"bibtype\":\"article\",\"author\":\"Liew, Chu Kong and Simpson, Raina J Y and Kwan, Ann H Y and Crofts, Linda A. and Loughlin, Fionna E. and Matthews, Jacqueline M. and Crossley, Merlin and Mackay, Joel P.\",\"doi\":\"10.1073/pnas.0407511102\",\"journal\":\"Proceedings of the National Academy of Sciences\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/Friend of GATA interaction},\\n type = {article},\\n year = {2005},\\n keywords = {Factor,Gene expression,Transcription},\\n pages = {583-588},\\n volume = {102},\\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/15644435},\\n month = {1},\\n day = {18},\\n id = {4fa93b4d-9743-397c-8bc8-7dec216c8aa7},\\n created = {2020-12-17T05:29:53.577Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:53.577Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Liew2005},\\n source_type = {ARTICLE},\\n notes = {cited By 65},\\n private_publication = {false},\\n abstract = {GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.},\\n bibtype = {article},\\n author = {Liew, Chu Kong and Simpson, Raina J Y and Kwan, Ann H Y and Crofts, Linda A. and Loughlin, Fionna E. and Matthews, Jacqueline M. and Crossley, Merlin and Mackay, Joel P.},\\n doi = {10.1073/pnas.0407511102},\\n journal = {Proceedings of the National Academy of Sciences},\\n number = {3}\\n}\",\"author_short\":[\"Liew,  C., K.\",\"Simpson,  R., J., Y.\",\"Kwan,  A., H., Y.\",\"Crofts,  L., A.\",\"Loughlin,  F., E.\",\"Matthews,  J., M.\",\"Crossley,  M.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://www.ncbi.nlm.nih.gov/pubmed/15644435\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005\",\"role\":\"author\",\"keyword\":[\"Factor\",\"Gene expression\",\"Transcription\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"DNA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II\",\"type\":\"article\",\"year\":\"1997\",\"keywords\":\"CD,DNA chemical footprinting,Fotemiistine,Intercalation,Linear dichroism,RNA polymerase II,SPXX peptides,YSPTSPSY,β-tum\",\"pages\":\"387-398\",\"volume\":\"42\",\"websites\":\"https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M\",\"month\":\"10\",\"day\":\"5\",\"id\":\"987f67ee-49d8-3803-8df7-9251cdbaf6c9\",\"created\":\"2020-12-17T05:29:53.650Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:53.650Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Harding1997\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 5\",\"private_publication\":false,\"abstract\":\"The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley & Sons, Inc.\",\"bibtype\":\"article\",\"author\":\"Harding, Margaret M. and Krippner, Guy Y. and Shelton, Cathryn J. and Rodger, Alison and Sanders, Karen J. and Mackay, Joel P. and Prakash, Arungundrum S.\",\"doi\":\"10.1002/(SICI)1097-0282(19971005)42:4<387::AID-BIP2>3.0.CO;2-M\",\"journal\":\"Biopolymers\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {DNA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II},\\n type = {article},\\n year = {1997},\\n keywords = {CD,DNA chemical footprinting,Fotemiistine,Intercalation,Linear dichroism,RNA polymerase II,SPXX peptides,YSPTSPSY,β-tum},\\n pages = {387-398},\\n volume = {42},\\n websites = {https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M},\\n month = {10},\\n day = {5},\\n id = {987f67ee-49d8-3803-8df7-9251cdbaf6c9},\\n created = {2020-12-17T05:29:53.650Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:53.650Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Harding1997},\\n source_type = {ARTICLE},\\n notes = {cited By 5},\\n private_publication = {false},\\n abstract = {The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley & Sons, Inc.},\\n bibtype = {article},\\n author = {Harding, Margaret M. and Krippner, Guy Y. and Shelton, Cathryn J. and Rodger, Alison and Sanders, Karen J. and Mackay, Joel P. and Prakash, Arungundrum S.},\\n doi = {10.1002/(SICI)1097-0282(19971005)42:4<387::AID-BIP2>3.0.CO;2-M},\\n journal = {Biopolymers},\\n number = {4}\\n}\",\"author_short\":[\"Harding,  M., M.\",\"Krippner,  G., Y.\",\"Shelton,  C., J.\",\"Rodger,  A.\",\"Sanders,  K., J.\",\"Mackay,  J., P.\",\"Prakash,  A., S.\"],\"urls\":{\"Website\":\"https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"harding-krippner-shelton-rodger-sanders-mackay-prakash-dnabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997\",\"role\":\"author\",\"keyword\":[\"CD\",\"DNA chemical footprinting\",\"Fotemiistine\",\"Intercalation\",\"Linear dichroism\",\"RNA polymerase II\",\"SPXX peptides\",\"YSPTSPSY\",\"β-tum\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry\",\"type\":\"article\",\"year\":\"2012\",\"keywords\":\"Mycobacteria,PIN-domain,RNA interferase,RNase,Toxin-antitoxin,VapC\",\"pages\":\"1267-1278\",\"volume\":\"18\",\"websites\":\"http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111\",\"month\":\"6\",\"day\":\"1\",\"id\":\"52a8f59e-942d-361e-8a56-5a3ea03f061e\",\"created\":\"2020-12-17T05:29:53.840Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:53.840Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"McKenzie2012\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 31\",\"private_publication\":false,\"abstract\":\"The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.\",\"bibtype\":\"article\",\"author\":\"McKenzie, J. L. and Duyvestyn, Johanna M. and Smith, Tony and Bendak, Katerina and MacKay, J. and Cursons, R. and Cook, Gregory M. and Arcus, Vickery L.\",\"doi\":\"10.1261/rna.031229.111\",\"journal\":\"RNA\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry},\\n type = {article},\\n year = {2012},\\n keywords = {Mycobacteria,PIN-domain,RNA interferase,RNase,Toxin-antitoxin,VapC},\\n pages = {1267-1278},\\n volume = {18},\\n websites = {http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111},\\n month = {6},\\n day = {1},\\n id = {52a8f59e-942d-361e-8a56-5a3ea03f061e},\\n created = {2020-12-17T05:29:53.840Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:53.840Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {McKenzie2012},\\n source_type = {ARTICLE},\\n notes = {cited By 31},\\n private_publication = {false},\\n abstract = {The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.},\\n bibtype = {article},\\n author = {McKenzie, J. L. and Duyvestyn, Johanna M. and Smith, Tony and Bendak, Katerina and MacKay, J. and Cursons, R. and Cook, Gregory M. and Arcus, Vickery L.},\\n doi = {10.1261/rna.031229.111},\\n journal = {RNA},\\n number = {6}\\n}\",\"author_short\":[\"McKenzie,  J., L.\",\"Duyvestyn,  J., M.\",\"Smith,  T.\",\"Bendak,  K.\",\"MacKay,  J.\",\"Cursons,  R.\",\"Cook,  G., M.\",\"Arcus,  V., L.\"],\"urls\":{\"Website\":\"http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012\",\"role\":\"author\",\"keyword\":[\"Mycobacteria\",\"PIN-domain\",\"RNA interferase\",\"RNase\",\"Toxin-antitoxin\",\"VapC\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Interdomain orientation of cardiac Troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state\",\"type\":\"article\",\"year\":\"2012\",\"keywords\":\"Cardiac troponin C,Ensemble states,Paramagnetic relaxation enhancement,Site-directed spin labeling,Solution NMR\",\"pages\":\"1376-1387\",\"volume\":\"21\",\"websites\":\"http://doi.wiley.com/10.1002/pro.2124\",\"month\":\"9\",\"id\":\"136eec2a-03b6-3cd9-bce5-f2eed2bdbc99\",\"created\":\"2020-12-17T05:29:54.085Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:54.085Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Cordina2012\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 9\",\"private_publication\":false,\"abstract\":\"Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.\",\"bibtype\":\"article\",\"author\":\"Cordina, Nicole M. and Liew, Chu Kong and Gell, David A. and Fajer, Piotr G. and Mackay, Joel P. and Brown, Louise J.\",\"doi\":\"10.1002/pro.2124\",\"journal\":\"Protein Science\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {Interdomain orientation of cardiac Troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state},\\n type = {article},\\n year = {2012},\\n keywords = {Cardiac troponin C,Ensemble states,Paramagnetic relaxation enhancement,Site-directed spin labeling,Solution NMR},\\n pages = {1376-1387},\\n volume = {21},\\n websites = {http://doi.wiley.com/10.1002/pro.2124},\\n month = {9},\\n id = {136eec2a-03b6-3cd9-bce5-f2eed2bdbc99},\\n created = {2020-12-17T05:29:54.085Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:54.085Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Cordina2012},\\n source_type = {ARTICLE},\\n notes = {cited By 9},\\n private_publication = {false},\\n abstract = {Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.},\\n bibtype = {article},\\n author = {Cordina, Nicole M. and Liew, Chu Kong and Gell, David A. and Fajer, Piotr G. and Mackay, Joel P. and Brown, Louise J.},\\n doi = {10.1002/pro.2124},\\n journal = {Protein Science},\\n number = {9}\\n}\",\"author_short\":[\"Cordina,  N., M.\",\"Liew,  C., K.\",\"Gell,  D., A.\",\"Fajer,  P., G.\",\"Mackay,  J., P.\",\"Brown,  L., J.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1002/pro.2124\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012\",\"role\":\"author\",\"keyword\":[\"Cardiac troponin C\",\"Ensemble states\",\"Paramagnetic relaxation enhancement\",\"Site-directed spin labeling\",\"Solution NMR\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"type\":\"inbook\",\"year\":\"2010\",\"keywords\":\"Coupled folding and binding,Hotspots,Post-translational modifications,Protein interfaces,Protein recognition\",\"pages\":\"505-532\",\"volume\":\"2\",\"websites\":\"http://doi.wiley.com/10.1002/9783527631780.ch12\",\"month\":\"12\",\"publisher\":\"Wiley-VCH Verlag GmbH & Co. KGaA\",\"day\":\"28\",\"city\":\"Weinheim, Germany\",\"id\":\"31798292-fed8-39f9-8349-947a9649059a\",\"created\":\"2020-12-17T05:29:54.383Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:54.383Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Mansfield2010\",\"source_type\":\"BOOK\",\"notes\":\"cited By 0\",\"private_publication\":false,\"bibtype\":\"inbook\",\"author\":\"Mansfield, Robyn E. and Cross, Arwen J. and Matthews, Jacqueline M. and Mackay, Joel P.\",\"doi\":\"10.1002/9783527631780.ch12\",\"chapter\":\"Protein Recognition\",\"title\":\"Amino Acids, Peptides and Proteins in Organic Chemistry\",\"bibtex\":\"@inbook{\\n type = {inbook},\\n year = {2010},\\n keywords = {Coupled folding and binding,Hotspots,Post-translational modifications,Protein interfaces,Protein recognition},\\n pages = {505-532},\\n volume = {2},\\n websites = {http://doi.wiley.com/10.1002/9783527631780.ch12},\\n month = {12},\\n publisher = {Wiley-VCH Verlag GmbH & Co. KGaA},\\n day = {28},\\n city = {Weinheim, Germany},\\n id = {31798292-fed8-39f9-8349-947a9649059a},\\n created = {2020-12-17T05:29:54.383Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:54.383Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Mansfield2010},\\n source_type = {BOOK},\\n notes = {cited By 0},\\n private_publication = {false},\\n bibtype = {inbook},\\n author = {Mansfield, Robyn E. and Cross, Arwen J. and Matthews, Jacqueline M. and Mackay, Joel P.},\\n doi = {10.1002/9783527631780.ch12},\\n chapter = {Protein Recognition},\\n title = {Amino Acids, Peptides and Proteins in Organic Chemistry}\\n}\",\"author_short\":[\"Mansfield,  R., E.\",\"Cross,  A., J.\",\"Matthews,  J., M.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1002/9783527631780.ch12\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mansfield-cross-matthews-mackay-aminoacidspeptidesandproteinsinorganicchemistry-2010\",\"role\":\"author\",\"keyword\":[\"Coupled folding and binding\",\"Hotspots\",\"Post-translational modifications\",\"Protein interfaces\",\"Protein recognition\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications\",\"type\":\"article\",\"year\":\"2009\",\"keywords\":\"Chromodomain helicase DNA-binding protein 4 (CHD4),Histone,Methylation,Plant homeodomain (PHD)\",\"pages\":\"179-187\",\"volume\":\"423\",\"websites\":\"https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is\",\"month\":\"10\",\"day\":\"15\",\"id\":\"4c2da44b-b1f6-3dcb-a479-0d561aabb87c\",\"created\":\"2020-12-17T05:29:54.740Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:54.740Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Musselman2009\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 73\",\"private_publication\":false,\"abstract\":\"CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation or methylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2–H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin.\",\"bibtype\":\"article\",\"author\":\"Musselman, Catherine A. and Mansfield, Robyn E. and Garske, Adam L. and Davrazou, Foteini and Kwan, Ann H. and Oliver, Samuel S. and O'Leary, Heather and Denu, John M. and Mackay, Joel P. and Kutateladze, Tatiana G.\",\"doi\":\"10.1042/BJ20090870\",\"journal\":\"Biochemical Journal\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications},\\n type = {article},\\n year = {2009},\\n keywords = {Chromodomain helicase DNA-binding protein 4 (CHD4),Histone,Methylation,Plant homeodomain (PHD)},\\n pages = {179-187},\\n volume = {423},\\n websites = {https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is},\\n month = {10},\\n day = {15},\\n id = {4c2da44b-b1f6-3dcb-a479-0d561aabb87c},\\n created = {2020-12-17T05:29:54.740Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:54.740Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Musselman2009},\\n source_type = {ARTICLE},\\n notes = {cited By 73},\\n private_publication = {false},\\n abstract = {CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation or methylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2–H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin.},\\n bibtype = {article},\\n author = {Musselman, Catherine A. and Mansfield, Robyn E. and Garske, Adam L. and Davrazou, Foteini and Kwan, Ann H. and Oliver, Samuel S. and O'Leary, Heather and Denu, John M. and Mackay, Joel P. and Kutateladze, Tatiana G.},\\n doi = {10.1042/BJ20090870},\\n journal = {Biochemical Journal},\\n number = {2}\\n}\",\"author_short\":[\"Musselman,  C., A.\",\"Mansfield,  R., E.\",\"Garske,  A., L.\",\"Davrazou,  F.\",\"Kwan,  A., H.\",\"Oliver,  S., S.\",\"O'Leary,  H.\",\"Denu,  J., M.\",\"Mackay,  J., P.\",\"Kutateladze,  T., G.\"],\"urls\":{\"Website\":\"https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009\",\"role\":\"author\",\"keyword\":[\"Chromodomain helicase DNA-binding protein 4 (CHD4)\",\"Histone\",\"Methylation\",\"Plant homeodomain (PHD)\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The structural analysis of proteinâ&#128;&#147;protein interactions by NMR spectroscopy\",\"type\":\"article\",\"year\":\"2009\",\"keywords\":\"NMR spectroscopy,Protein-protein interactions,Structural biology,Technology\",\"pages\":\"5224-5232\",\"volume\":\"9\",\"websites\":\"http://doi.wiley.com/10.1002/pmic.200900303\",\"month\":\"12\",\"id\":\"fc390463-14cb-327f-93bf-fbec6b3cd8fd\",\"created\":\"2020-12-17T05:29:55.049Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:55.049Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"OConnell2009\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 44\",\"private_publication\":false,\"abstract\":\"A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.\",\"bibtype\":\"article\",\"author\":\"O'Connell, Mitchell R. and Gamsjaeger, Roland and Mackay, Joel P.\",\"doi\":\"10.1002/pmic.200900303\",\"journal\":\"PROTEOMICS\",\"number\":\"23\",\"bibtex\":\"@article{\\n title = {The structural analysis of proteinâ&#128;&#147;protein interactions by NMR spectroscopy},\\n type = {article},\\n year = {2009},\\n keywords = {NMR spectroscopy,Protein-protein interactions,Structural biology,Technology},\\n pages = {5224-5232},\\n volume = {9},\\n websites = {http://doi.wiley.com/10.1002/pmic.200900303},\\n month = {12},\\n id = {fc390463-14cb-327f-93bf-fbec6b3cd8fd},\\n created = {2020-12-17T05:29:55.049Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:55.049Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {OConnell2009},\\n source_type = {ARTICLE},\\n notes = {cited By 44},\\n private_publication = {false},\\n abstract = {A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.},\\n bibtype = {article},\\n author = {O'Connell, Mitchell R. and Gamsjaeger, Roland and Mackay, Joel P.},\\n doi = {10.1002/pmic.200900303},\\n journal = {PROTEOMICS},\\n number = {23}\\n}\",\"author_short\":[\"O'Connell,  M., R.\",\"Gamsjaeger,  R.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1002/pmic.200900303\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"oconnell-gamsjaeger-mackay-thestructuralanalysisofprotein128147proteininteractionsbynmrspectroscopy-2009\",\"role\":\"author\",\"keyword\":[\"NMR spectroscopy\",\"Protein-protein interactions\",\"Structural biology\",\"Technology\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana\",\"type\":\"article\",\"year\":\"2018\",\"keywords\":\"DNA binding affinity,DOF zinc finger domain,Gel retardation assay,Microscale thermophoresis\",\"pages\":\"167-176\",\"volume\":\"8\",\"websites\":\"http://bi.tbzmed.ac.ir/Abstract/bi-17551\",\"month\":\"1\",\"day\":\"28\",\"id\":\"dd4675dd-58e2-3545-8030-ebe4c5b425a8\",\"created\":\"2020-12-17T05:29:55.192Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:55.192Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"MoghaddasSani2018\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 1\",\"private_publication\":false,\"abstract\":\"Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.\",\"bibtype\":\"article\",\"author\":\"Moghaddas Sani, Hakimeh and Hamzeh-Mivehroud, Maryam and Silva, Ana P. and Walshe, James L. and Mohammadi, S. Abolghasem and Rahbar-Shahrouziasl, Mahdyieh and Abbasi, Milad and Jamshidi, Omid and Low, Jason K.K. and Dastmalchi, Siavoush and Mackay, Joel P.\",\"doi\":\"10.15171/bi.2018.19\",\"journal\":\"BioImpacts\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana},\\n type = {article},\\n year = {2018},\\n keywords = {DNA binding affinity,DOF zinc finger domain,Gel retardation assay,Microscale thermophoresis},\\n pages = {167-176},\\n volume = {8},\\n websites = {http://bi.tbzmed.ac.ir/Abstract/bi-17551},\\n month = {1},\\n day = {28},\\n id = {dd4675dd-58e2-3545-8030-ebe4c5b425a8},\\n created = {2020-12-17T05:29:55.192Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:55.192Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {MoghaddasSani2018},\\n source_type = {ARTICLE},\\n notes = {cited By 1},\\n private_publication = {false},\\n abstract = {Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.},\\n bibtype = {article},\\n author = {Moghaddas Sani, Hakimeh and Hamzeh-Mivehroud, Maryam and Silva, Ana P. and Walshe, James L. and Mohammadi, S. Abolghasem and Rahbar-Shahrouziasl, Mahdyieh and Abbasi, Milad and Jamshidi, Omid and Low, Jason K.K. and Dastmalchi, Siavoush and Mackay, Joel P.},\\n doi = {10.15171/bi.2018.19},\\n journal = {BioImpacts},\\n number = {3}\\n}\",\"author_short\":[\"Moghaddas Sani,  H.\",\"Hamzeh-Mivehroud,  M.\",\"Silva,  A., P.\",\"Walshe,  J., L.\",\"Mohammadi,  S., A.\",\"Rahbar-Shahrouziasl,  M.\",\"Abbasi,  M.\",\"Jamshidi,  O.\",\"Low,  J., K.\",\"Dastmalchi,  S.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://bi.tbzmed.ac.ir/Abstract/bi-17551\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"moghaddassani-hamzehmivehroud-silva-walshe-mohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018\",\"role\":\"author\",\"keyword\":[\"DNA binding affinity\",\"DOF zinc finger domain\",\"Gel retardation assay\",\"Microscale thermophoresis\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Presence of transient helical segments in the galanin-like peptide evident from 1H NMR, circular dichroism, and prediction studies\",\"type\":\"article\",\"year\":\"2004\",\"keywords\":\"Circular dichroism,Galanin-like peptide,NMR spectroscopy,Nascent helix,Secondary structure prediction\",\"pages\":\"261-271\",\"volume\":\"146\",\"websites\":\"http://www.ncbi.nlm.nih.gov/pubmed/15099568\",\"month\":\"6\",\"id\":\"ccd45448-027a-3ba1-94ee-88d2de266d92\",\"created\":\"2020-12-17T05:29:55.417Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:55.417Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Dastmalchi2004\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 7\",\"private_publication\":false,\"abstract\":\"Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods.\",\"bibtype\":\"article\",\"author\":\"Dastmalchi, Siavoush and Church, W.Bret and Morris, Michael B. and Iismaa, Tiina P. and Mackay, Joel P.\",\"doi\":\"10.1016/j.jsb.2004.01.004\",\"journal\":\"Journal of Structural Biology\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Presence of transient helical segments in the galanin-like peptide evident from 1H NMR, circular dichroism, and prediction studies},\\n type = {article},\\n year = {2004},\\n keywords = {Circular dichroism,Galanin-like peptide,NMR spectroscopy,Nascent helix,Secondary structure prediction},\\n pages = {261-271},\\n volume = {146},\\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/15099568},\\n month = {6},\\n id = {ccd45448-027a-3ba1-94ee-88d2de266d92},\\n created = {2020-12-17T05:29:55.417Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:55.417Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Dastmalchi2004},\\n source_type = {ARTICLE},\\n notes = {cited By 7},\\n private_publication = {false},\\n abstract = {Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods.},\\n bibtype = {article},\\n author = {Dastmalchi, Siavoush and Church, W.Bret and Morris, Michael B. and Iismaa, Tiina P. and Mackay, Joel P.},\\n doi = {10.1016/j.jsb.2004.01.004},\\n journal = {Journal of Structural Biology},\\n number = {3}\\n}\",\"author_short\":[\"Dastmalchi,  S.\",\"Church,  W.\",\"Morris,  M., B.\",\"Iismaa,  T., P.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://www.ncbi.nlm.nih.gov/pubmed/15099568\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfrom1hnmrcirculardichroismandpredictionstudies-2004\",\"role\":\"author\",\"keyword\":[\"Circular dichroism\",\"Galanin-like peptide\",\"NMR spectroscopy\",\"Nascent helix\",\"Secondary structure prediction\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1\",\"type\":\"article\",\"year\":\"2011\",\"keywords\":\"Haematopoiesis,Protein-DNA interactions,Protein-protein interactions,Transcription factor complex\",\"pages\":\"14443-14448\",\"volume\":\"108\",\"websites\":\"http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108\",\"month\":\"8\",\"day\":\"30\",\"id\":\"a786b1a3-bd30-3c66-8aff-7ffb5cbbb0da\",\"created\":\"2020-12-17T05:29:55.512Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:55.512Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Wilkinson-White2011\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 24\",\"private_publication\":false,\"abstract\":\"The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (<50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.\",\"bibtype\":\"article\",\"author\":\"Wilkinson-White, Lorna and Gamsjaeger, Roland and Dastmalchi, Siavoush and Wienert, Beeke and Stokes, Philippa H. and Crossley, Merlin and Mackay, Joel P. and Matthews, Jacqueline M.\",\"doi\":\"10.1073/pnas.1105898108\",\"journal\":\"Proceedings of the National Academy of Sciences\",\"number\":\"35\",\"bibtex\":\"@article{\\n title = {Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1},\\n type = {article},\\n year = {2011},\\n keywords = {Haematopoiesis,Protein-DNA interactions,Protein-protein interactions,Transcription factor complex},\\n pages = {14443-14448},\\n volume = {108},\\n websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108},\\n month = {8},\\n day = {30},\\n id = {a786b1a3-bd30-3c66-8aff-7ffb5cbbb0da},\\n created = {2020-12-17T05:29:55.512Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:55.512Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Wilkinson-White2011},\\n source_type = {ARTICLE},\\n notes = {cited By 24},\\n private_publication = {false},\\n abstract = {The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (<50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.},\\n bibtype = {article},\\n author = {Wilkinson-White, Lorna and Gamsjaeger, Roland and Dastmalchi, Siavoush and Wienert, Beeke and Stokes, Philippa H. and Crossley, Merlin and Mackay, Joel P. and Matthews, Jacqueline M.},\\n doi = {10.1073/pnas.1105898108},\\n journal = {Proceedings of the National Academy of Sciences},\\n number = {35}\\n}\",\"author_short\":[\"Wilkinson-White,  L.\",\"Gamsjaeger,  R.\",\"Dastmalchi,  S.\",\"Wienert,  B.\",\"Stokes,  P., H.\",\"Crossley,  M.\",\"Mackay,  J., P.\",\"Matthews,  J., M.\"],\"urls\":{\"Website\":\"http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011\",\"role\":\"author\",\"keyword\":[\"Haematopoiesis\",\"Protein-DNA interactions\",\"Protein-protein interactions\",\"Transcription factor complex\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Peppy: A virtual reality environment for exploring the principles of polypeptide structure\",\"type\":\"article\",\"year\":\"2020\",\"keywords\":\"polypeptide,protein,secondary structure,teaching,undergraduate,virtual reality\",\"pages\":\"157-168\",\"volume\":\"29\",\"websites\":\"http://www.ncbi.nlm.nih.gov/pubmed/31622516\",\"month\":\"1\",\"day\":\"11\",\"id\":\"fe01e53b-d05d-3939-8395-e8da99c6004e\",\"created\":\"2020-12-17T05:29:55.556Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:55.556Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Doak2020\",\"source_type\":\"article\",\"private_publication\":false,\"abstract\":\"A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.\",\"bibtype\":\"article\",\"author\":\"Doak, David G. and Denyer, Gareth S. and Gerrard, Juliet A. and Mackay, Joel P. and Allison, Jane R.\",\"doi\":\"10.1002/pro.3752\",\"journal\":\"Protein Science\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Peppy: A virtual reality environment for exploring the principles of polypeptide structure},\\n type = {article},\\n year = {2020},\\n keywords = {polypeptide,protein,secondary structure,teaching,undergraduate,virtual reality},\\n pages = {157-168},\\n volume = {29},\\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/31622516},\\n month = {1},\\n day = {11},\\n id = {fe01e53b-d05d-3939-8395-e8da99c6004e},\\n created = {2020-12-17T05:29:55.556Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:55.556Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Doak2020},\\n source_type = {article},\\n private_publication = {false},\\n abstract = {A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.},\\n bibtype = {article},\\n author = {Doak, David G. and Denyer, Gareth S. and Gerrard, Juliet A. and Mackay, Joel P. and Allison, Jane R.},\\n doi = {10.1002/pro.3752},\\n journal = {Protein Science},\\n number = {1}\\n}\",\"author_short\":[\"Doak,  D., G.\",\"Denyer,  G., S.\",\"Gerrard,  J., A.\",\"Mackay,  J., P.\",\"Allison,  J., R.\"],\"urls\":{\"Website\":\"http://www.ncbi.nlm.nih.gov/pubmed/31622516\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020\",\"role\":\"author\",\"keyword\":[\"polypeptide\",\"protein\",\"secondary structure\",\"teaching\",\"undergraduate\",\"virtual reality\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Engineering Specificity Changes on a RanBP2 Zinc Finger that Binds Single-Stranded RNA\",\"type\":\"article\",\"year\":\"2014\",\"keywords\":\"RNA binding,combinatorial chemistry,phage display,protein design,zinc fingers\",\"pages\":\"7848-7852\",\"volume\":\"53\",\"websites\":\"http://doi.wiley.com/10.1002/anie.201402980\",\"month\":\"7\",\"day\":\"21\",\"id\":\"5c546e1a-664d-3c49-b780-ca530f662b55\",\"created\":\"2020-12-17T05:29:55.706Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:55.706Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Vandevenne2014\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 3\",\"private_publication\":false,\"abstract\":\"The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.\",\"bibtype\":\"article\",\"author\":\"Vandevenne, Marylène and O'Connell, Mitchell R. and Helder, Stephanie and Shepherd, Nicholas E. and Matthews, Jacqueline M. and Kwan, Ann H. and Segal, David J. and Mackay, Joel P.\",\"doi\":\"10.1002/anie.201402980\",\"journal\":\"Angewandte Chemie International Edition\",\"number\":\"30\",\"bibtex\":\"@article{\\n title = {Engineering Specificity Changes on a RanBP2 Zinc Finger that Binds Single-Stranded RNA},\\n type = {article},\\n year = {2014},\\n keywords = {RNA binding,combinatorial chemistry,phage display,protein design,zinc fingers},\\n pages = {7848-7852},\\n volume = {53},\\n websites = {http://doi.wiley.com/10.1002/anie.201402980},\\n month = {7},\\n day = {21},\\n id = {5c546e1a-664d-3c49-b780-ca530f662b55},\\n created = {2020-12-17T05:29:55.706Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:55.706Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Vandevenne2014},\\n source_type = {ARTICLE},\\n notes = {cited By 3},\\n private_publication = {false},\\n abstract = {The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.},\\n bibtype = {article},\\n author = {Vandevenne, Marylène and O'Connell, Mitchell R. and Helder, Stephanie and Shepherd, Nicholas E. and Matthews, Jacqueline M. and Kwan, Ann H. and Segal, David J. and Mackay, Joel P.},\\n doi = {10.1002/anie.201402980},\\n journal = {Angewandte Chemie International Edition},\\n number = {30}\\n}\",\"author_short\":[\"Vandevenne,  M.\",\"O'Connell,  M., R.\",\"Helder,  S.\",\"Shepherd,  N., E.\",\"Matthews,  J., M.\",\"Kwan,  A., H.\",\"Segal,  D., J.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1002/anie.201402980\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014\",\"role\":\"author\",\"keyword\":[\"RNA binding\",\"combinatorial chemistry\",\"phage display\",\"protein design\",\"zinc fingers\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein\",\"type\":\"article\",\"year\":\"2016\",\"keywords\":\"RNA binding,RNA polymerase,SELEX,Spt4/5,transcription elongation,transcription elongation factor\",\"pages\":\"1710-1721\",\"volume\":\"25\",\"websites\":\"http://doi.wiley.com/10.1002/pro.2976\",\"month\":\"9\",\"id\":\"cb1f5362-a6b4-3c3e-a896-fd523c0b1404\",\"created\":\"2020-12-17T05:29:56.143Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:56.143Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Blythe2016\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 2\",\"private_publication\":false,\"abstract\":\"The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.\",\"bibtype\":\"article\",\"author\":\"Blythe, Amanda J. and Yazar-Klosinski, Berra and Webster, Michael W. and Chen, Eefei and Vandevenne, Marylène and Bendak, Katerina and Mackay, Joel P. and Hartzog, Grant A. and Vrielink, Alice\",\"doi\":\"10.1002/pro.2976\",\"journal\":\"Protein Science\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein},\\n type = {article},\\n year = {2016},\\n keywords = {RNA binding,RNA polymerase,SELEX,Spt4/5,transcription elongation,transcription elongation factor},\\n pages = {1710-1721},\\n volume = {25},\\n websites = {http://doi.wiley.com/10.1002/pro.2976},\\n month = {9},\\n id = {cb1f5362-a6b4-3c3e-a896-fd523c0b1404},\\n created = {2020-12-17T05:29:56.143Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:56.143Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Blythe2016},\\n source_type = {ARTICLE},\\n notes = {cited By 2},\\n private_publication = {false},\\n abstract = {The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.},\\n bibtype = {article},\\n author = {Blythe, Amanda J. and Yazar-Klosinski, Berra and Webster, Michael W. and Chen, Eefei and Vandevenne, Marylène and Bendak, Katerina and Mackay, Joel P. and Hartzog, Grant A. and Vrielink, Alice},\\n doi = {10.1002/pro.2976},\\n journal = {Protein Science},\\n number = {9}\\n}\",\"author_short\":[\"Blythe,  A., J.\",\"Yazar-Klosinski,  B.\",\"Webster,  M., W.\",\"Chen,  E.\",\"Vandevenne,  M.\",\"Bendak,  K.\",\"Mackay,  J., P.\",\"Hartzog,  G., A.\",\"Vrielink,  A.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1002/pro.2976\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016\",\"role\":\"author\",\"keyword\":[\"RNA binding\",\"RNA polymerase\",\"SELEX\",\"Spt4/5\",\"transcription elongation\",\"transcription elongation factor\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Backbone and sidechain 1H, 13C and 15N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans\",\"type\":\"article\",\"year\":\"2012\",\"keywords\":\"DewA,Functional amyloid,Hydrophobin,NMR assignment\",\"pages\":\"83-86\",\"volume\":\"6\",\"websites\":\"http://link.springer.com/10.1007/s12104-011-9330-5\",\"month\":\"4\",\"day\":\"4\",\"id\":\"852baf30-6704-30e4-8fdc-8cbe794bb64a\",\"created\":\"2020-12-17T05:29:56.237Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:56.237Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Morris2012\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 4\",\"private_publication\":false,\"abstract\":\"Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.\",\"bibtype\":\"article\",\"author\":\"Morris, Vanessa K. and Kwan, Ann H. and Mackay, Joel P. and Sunde, Margaret\",\"doi\":\"10.1007/s12104-011-9330-5\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Backbone and sidechain 1H, 13C and 15N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans},\\n type = {article},\\n year = {2012},\\n keywords = {DewA,Functional amyloid,Hydrophobin,NMR assignment},\\n pages = {83-86},\\n volume = {6},\\n websites = {http://link.springer.com/10.1007/s12104-011-9330-5},\\n month = {4},\\n day = {4},\\n id = {852baf30-6704-30e4-8fdc-8cbe794bb64a},\\n created = {2020-12-17T05:29:56.237Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:56.237Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Morris2012},\\n source_type = {ARTICLE},\\n notes = {cited By 4},\\n private_publication = {false},\\n abstract = {Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.},\\n bibtype = {article},\\n author = {Morris, Vanessa K. and Kwan, Ann H. and Mackay, Joel P. and Sunde, Margaret},\\n doi = {10.1007/s12104-011-9330-5},\\n journal = {Biomolecular NMR Assignments},\\n number = {1}\\n}\",\"author_short\":[\"Morris,  V., K.\",\"Kwan,  A., H.\",\"Mackay,  J., P.\",\"Sunde,  M.\"],\"urls\":{\"Website\":\"http://link.springer.com/10.1007/s12104-011-9330-5\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"morris-kwan-mackay-sunde-backboneandsidechain1h13cand15nchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012\",\"role\":\"author\",\"keyword\":[\"DewA\",\"Functional amyloid\",\"Hydrophobin\",\"NMR assignment\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Ikaros: a key regulator of haematopoiesis\",\"type\":\"article\",\"year\":\"2002\",\"keywords\":\"Haematopoiesis,Ikaros,Pericentromeric-heterochromatin,Transcription\",\"pages\":\"1304-1307\",\"volume\":\"34\",\"websites\":\"http://www.ncbi.nlm.nih.gov/pubmed/12127581\",\"month\":\"10\",\"id\":\"5eef5524-c104-30d5-8fd5-93d7f32bc139\",\"created\":\"2020-12-17T05:29:56.237Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:56.237Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Westman2002\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 21\",\"private_publication\":false,\"abstract\":\"Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function--unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription.\",\"bibtype\":\"article\",\"author\":\"Westman, Belinda J. and Mackay, Joel P. and Gell, David\",\"doi\":\"10.1016/S1357-2725(02)00070-5\",\"journal\":\"The International Journal of Biochemistry & Cell Biology\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {Ikaros: a key regulator of haematopoiesis},\\n type = {article},\\n year = {2002},\\n keywords = {Haematopoiesis,Ikaros,Pericentromeric-heterochromatin,Transcription},\\n pages = {1304-1307},\\n volume = {34},\\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/12127581},\\n month = {10},\\n id = {5eef5524-c104-30d5-8fd5-93d7f32bc139},\\n created = {2020-12-17T05:29:56.237Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:56.237Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Westman2002},\\n source_type = {ARTICLE},\\n notes = {cited By 21},\\n private_publication = {false},\\n abstract = {Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function--unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription.},\\n bibtype = {article},\\n author = {Westman, Belinda J. and Mackay, Joel P. and Gell, David},\\n doi = {10.1016/S1357-2725(02)00070-5},\\n journal = {The International Journal of Biochemistry & Cell Biology},\\n number = {10}\\n}\",\"author_short\":[\"Westman,  B., J.\",\"Mackay,  J., P.\",\"Gell,  D.\"],\"urls\":{\"Website\":\"http://www.ncbi.nlm.nih.gov/pubmed/12127581\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002\",\"role\":\"author\",\"keyword\":[\"Haematopoiesis\",\"Ikaros\",\"Pericentromeric-heterochromatin\",\"Transcription\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution structure of a tethered Lmo2 LIM2 /Ldb1 LID complex\",\"type\":\"article\",\"year\":\"2012\",\"keywords\":\"Ldb1,Lmo2,Modular binding,NMR structure\",\"pages\":\"1768-1774\",\"volume\":\"21\",\"websites\":\"http://doi.wiley.com/10.1002/pro.2153\",\"month\":\"11\",\"id\":\"574f81df-0620-3c4a-af49-474d48a6b829\",\"created\":\"2020-12-17T05:29:56.294Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:56.294Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Dastmalchi2012\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 7\",\"private_publication\":false,\"abstract\":\"LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.\",\"bibtype\":\"article\",\"author\":\"Dastmalchi, Siavoush and Wilkinson-White, Lorna and Kwan, Ann H. and Gamsjaeger, Roland and Mackay, Joel P. and Matthews, Jacqueline M.\",\"doi\":\"10.1002/pro.2153\",\"journal\":\"Protein Science\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Solution structure of a tethered Lmo2 LIM2 /Ldb1 LID complex},\\n type = {article},\\n year = {2012},\\n keywords = {Ldb1,Lmo2,Modular binding,NMR structure},\\n pages = {1768-1774},\\n volume = {21},\\n websites = {http://doi.wiley.com/10.1002/pro.2153},\\n month = {11},\\n id = {574f81df-0620-3c4a-af49-474d48a6b829},\\n created = {2020-12-17T05:29:56.294Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:56.294Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Dastmalchi2012},\\n source_type = {ARTICLE},\\n notes = {cited By 7},\\n private_publication = {false},\\n abstract = {LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.},\\n bibtype = {article},\\n author = {Dastmalchi, Siavoush and Wilkinson-White, Lorna and Kwan, Ann H. and Gamsjaeger, Roland and Mackay, Joel P. and Matthews, Jacqueline M.},\\n doi = {10.1002/pro.2153},\\n journal = {Protein Science},\\n number = {11}\\n}\",\"author_short\":[\"Dastmalchi,  S.\",\"Wilkinson-White,  L.\",\"Kwan,  A., H.\",\"Gamsjaeger,  R.\",\"Mackay,  J., P.\",\"Matthews,  J., M.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1002/pro.2153\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2lim2ldb1lidcomplex-2012\",\"role\":\"author\",\"keyword\":[\"Ldb1\",\"Lmo2\",\"Modular binding\",\"NMR structure\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells\",\"type\":\"article\",\"year\":\"2015\",\"keywords\":\"Affinity purification,Friend of GATA1 (FOG1),Multi-protein complex,Nucleosome remodeling and deacetylase complex pept\",\"pages\":\"960-965\",\"volume\":\"23\",\"websites\":\"https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395\",\"month\":\"3\",\"id\":\"c7a33bb8-06cd-300f-9f85-c79fbd926928\",\"created\":\"2020-12-17T05:29:56.497Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:56.497Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Saathoff2015\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 5\",\"private_publication\":false,\"abstract\":\"We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).\",\"bibtype\":\"article\",\"author\":\"Saathoff, Hinnerk and Brofelth, Mattias and Trinh, Anne and Parker, Benjamin L. and Ryan, Daniel P. and Low, Jason K.K. and Webb, Sarah R. and Silva, Ana P.G. and Mackay, Joel P. and Shepherd, Nicholas E.\",\"doi\":\"10.1016/j.bmc.2015.01.023\",\"journal\":\"Bioorganic & Medicinal Chemistry\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells},\\n type = {article},\\n year = {2015},\\n keywords = {Affinity purification,Friend of GATA1 (FOG1),Multi-protein complex,Nucleosome remodeling and deacetylase complex pept},\\n pages = {960-965},\\n volume = {23},\\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395},\\n month = {3},\\n id = {c7a33bb8-06cd-300f-9f85-c79fbd926928},\\n created = {2020-12-17T05:29:56.497Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:56.497Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Saathoff2015},\\n source_type = {ARTICLE},\\n notes = {cited By 5},\\n private_publication = {false},\\n abstract = {We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).},\\n bibtype = {article},\\n author = {Saathoff, Hinnerk and Brofelth, Mattias and Trinh, Anne and Parker, Benjamin L. and Ryan, Daniel P. and Low, Jason K.K. and Webb, Sarah R. and Silva, Ana P.G. and Mackay, Joel P. and Shepherd, Nicholas E.},\\n doi = {10.1016/j.bmc.2015.01.023},\\n journal = {Bioorganic & Medicinal Chemistry},\\n number = {5}\\n}\",\"author_short\":[\"Saathoff,  H.\",\"Brofelth,  M.\",\"Trinh,  A.\",\"Parker,  B., L.\",\"Ryan,  D., P.\",\"Low,  J., K.\",\"Webb,  S., R.\",\"Silva,  A., P.\",\"Mackay,  J., P.\",\"Shepherd,  N., E.\"],\"urls\":{\"Website\":\"https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015\",\"role\":\"author\",\"keyword\":[\"Affinity purification\",\"Friend of GATA1 (FOG1)\",\"Multi-protein complex\",\"Nucleosome remodeling and deacetylase complex pept\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Grb7-SH2 domain dimerisation is affected by a single point mutation.\",\"type\":\"article\",\"year\":\"2005\",\"keywords\":\"Analytical ultracentrifugation,Growth factor receptor bound protein 7,Protein engineering,Size-exclusion chromatography,Src homology 2 domain\",\"pages\":\"454-60\",\"volume\":\"34\",\"websites\":\"http://www.ncbi.nlm.nih.gov/pubmed/15841400\",\"month\":\"7\",\"day\":\"20\",\"id\":\"042bb7c3-136a-3d03-a241-4b3ebfa7ee45\",\"created\":\"2020-12-17T05:29:56.590Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:56.590Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Porter2005\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 20\",\"private_publication\":false,\"abstract\":\"Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 muM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation.\",\"bibtype\":\"article\",\"author\":\"Porter, Corrine J. and Wilce, Matthew C J and Mackay, Joel P. and Leedman, Peter and Wilce, Jackie A.\",\"doi\":\"10.1007/s00249-005-0480-1\",\"journal\":\"European biophysics journal : EBJ\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Grb7-SH2 domain dimerisation is affected by a single point mutation.},\\n type = {article},\\n year = {2005},\\n keywords = {Analytical ultracentrifugation,Growth factor receptor bound protein 7,Protein engineering,Size-exclusion chromatography,Src homology 2 domain},\\n pages = {454-60},\\n volume = {34},\\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/15841400},\\n month = {7},\\n day = {20},\\n id = {042bb7c3-136a-3d03-a241-4b3ebfa7ee45},\\n created = {2020-12-17T05:29:56.590Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:56.590Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Porter2005},\\n source_type = {ARTICLE},\\n notes = {cited By 20},\\n private_publication = {false},\\n abstract = {Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 muM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation.},\\n bibtype = {article},\\n author = {Porter, Corrine J. and Wilce, Matthew C J and Mackay, Joel P. and Leedman, Peter and Wilce, Jackie A.},\\n doi = {10.1007/s00249-005-0480-1},\\n journal = {European biophysics journal : EBJ},\\n number = {5}\\n}\",\"author_short\":[\"Porter,  C., J.\",\"Wilce,  M., C., J.\",\"Mackay,  J., P.\",\"Leedman,  P.\",\"Wilce,  J., A.\"],\"urls\":{\"Website\":\"http://www.ncbi.nlm.nih.gov/pubmed/15841400\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005\",\"role\":\"author\",\"keyword\":[\"Analytical ultracentrifugation\",\"Growth factor receptor bound protein 7\",\"Protein engineering\",\"Size-exclusion chromatography\",\"Src homology 2 domain\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"1H, 13C and 15N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis\",\"type\":\"article\",\"year\":\"2010\",\"keywords\":\"Bacillus subtilis,Bacterial signal transduction,Histidine kinase inhibition,KipI\",\"pages\":\"167-169\",\"volume\":\"4\",\"websites\":\"http://link.springer.com/10.1007/s12104-010-9237-6\",\"month\":\"10\",\"day\":\"4\",\"id\":\"48c78b62-9179-35cb-ae9d-83d16d7e9f3b\",\"created\":\"2020-12-17T05:29:56.688Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:56.688Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Hynson2010\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 2\",\"private_publication\":false,\"abstract\":\"KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.\",\"bibtype\":\"article\",\"author\":\"Hynson, Robert M. G. and Kwan, Ann H. and Jacques, David A. and Mackay, Joel P. and Trewhella, Jill\",\"doi\":\"10.1007/s12104-010-9237-6\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {1H, 13C and 15N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis},\\n type = {article},\\n year = {2010},\\n keywords = {Bacillus subtilis,Bacterial signal transduction,Histidine kinase inhibition,KipI},\\n pages = {167-169},\\n volume = {4},\\n websites = {http://link.springer.com/10.1007/s12104-010-9237-6},\\n month = {10},\\n day = {4},\\n id = {48c78b62-9179-35cb-ae9d-83d16d7e9f3b},\\n created = {2020-12-17T05:29:56.688Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:56.688Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Hynson2010},\\n source_type = {ARTICLE},\\n notes = {cited By 2},\\n private_publication = {false},\\n abstract = {KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.},\\n bibtype = {article},\\n author = {Hynson, Robert M. G. and Kwan, Ann H. and Jacques, David A. and Mackay, Joel P. and Trewhella, Jill},\\n doi = {10.1007/s12104-010-9237-6},\\n journal = {Biomolecular NMR Assignments},\\n number = {2}\\n}\",\"author_short\":[\"Hynson,  R., M., G.\",\"Kwan,  A., H.\",\"Jacques,  D., A.\",\"Mackay,  J., P.\",\"Trewhella,  J.\"],\"urls\":{\"Website\":\"http://link.springer.com/10.1007/s12104-010-9237-6\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"hynson-kwan-jacques-mackay-trewhella-1h13cand15nbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010\",\"role\":\"author\",\"keyword\":[\"Bacillus subtilis\",\"Bacterial signal transduction\",\"Histidine kinase inhibition\",\"KipI\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG.\",\"type\":\"article\",\"year\":\"1999\",\"keywords\":\"GATA-1,Protein-protein interactions,Solution structure,Zinc finger\",\"pages\":\"249-62\",\"volume\":\"13\",\"websites\":\"http://www.ncbi.nlm.nih.gov/pubmed/10212985\",\"month\":\"3\",\"id\":\"1bc4df8b-a093-3a22-ba88-996d8029dfd5\",\"created\":\"2020-12-17T05:29:56.853Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:56.853Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Kowalski1999\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 48\",\"private_publication\":false,\"abstract\":\"Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted beta-hairpins and a single alpha-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA-1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.\",\"bibtype\":\"article\",\"author\":\"Kowalski, K. and Czolij, R. and King, G. F. and Crossley, M. and Mackay, J. P.\",\"doi\":\"https://doi.org/10.1023/a:1008309602929\",\"journal\":\"Journal of Biomolecular NMR\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG.},\\n type = {article},\\n year = {1999},\\n keywords = {GATA-1,Protein-protein interactions,Solution structure,Zinc finger},\\n pages = {249-62},\\n volume = {13},\\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/10212985},\\n month = {3},\\n id = {1bc4df8b-a093-3a22-ba88-996d8029dfd5},\\n created = {2020-12-17T05:29:56.853Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:56.853Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Kowalski1999},\\n source_type = {ARTICLE},\\n notes = {cited By 48},\\n private_publication = {false},\\n abstract = {Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted beta-hairpins and a single alpha-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA-1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.},\\n bibtype = {article},\\n author = {Kowalski, K. and Czolij, R. and King, G. F. and Crossley, M. and Mackay, J. P.},\\n doi = {https://doi.org/10.1023/a:1008309602929},\\n journal = {Journal of Biomolecular NMR},\\n number = {3}\\n}\",\"author_short\":[\"Kowalski,  K.\",\"Czolij,  R.\",\"King,  G., F.\",\"Crossley,  M.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://www.ncbi.nlm.nih.gov/pubmed/10212985\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999\",\"role\":\"author\",\"keyword\":[\"GATA-1\",\"Protein-protein interactions\",\"Solution structure\",\"Zinc finger\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes\",\"type\":\"article\",\"year\":\"2011\",\"keywords\":\"Gene regulation,Hematopoiesis,Posttranslational modifications\",\"pages\":\"E159-E168\",\"volume\":\"108\",\"websites\":\"http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108\",\"month\":\"5\",\"day\":\"31\",\"id\":\"f5506c6c-a1c6-37df-8e6d-aee9893f4e46\",\"created\":\"2020-12-17T05:29:57.064Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:57.064Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Lamonica2011\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 98\",\"private_publication\":false,\"abstract\":\"Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 \\\"reads\\\" acetyl marks on nuclear factors to promote their stable association with chromatin.\",\"bibtype\":\"article\",\"author\":\"Lamonica, Janine M. and Deng, Wulan and Kadauke, Stephan and Campbell, Amy E. and Gamsjaeger, Roland and Wang, Hongxin and Cheng, Yong and Billin, Andrew N. and Hardison, Ross C. and Mackay, Joel P. and Blobel, Gerd A.\",\"doi\":\"10.1073/pnas.1102140108\",\"journal\":\"Proceedings of the National Academy of Sciences\",\"number\":\"22\",\"bibtex\":\"@article{\\n title = {Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes},\\n type = {article},\\n year = {2011},\\n keywords = {Gene regulation,Hematopoiesis,Posttranslational modifications},\\n pages = {E159-E168},\\n volume = {108},\\n websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108},\\n month = {5},\\n day = {31},\\n id = {f5506c6c-a1c6-37df-8e6d-aee9893f4e46},\\n created = {2020-12-17T05:29:57.064Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:57.064Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Lamonica2011},\\n source_type = {ARTICLE},\\n notes = {cited By 98},\\n private_publication = {false},\\n abstract = {Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 \\\"reads\\\" acetyl marks on nuclear factors to promote their stable association with chromatin.},\\n bibtype = {article},\\n author = {Lamonica, Janine M. and Deng, Wulan and Kadauke, Stephan and Campbell, Amy E. and Gamsjaeger, Roland and Wang, Hongxin and Cheng, Yong and Billin, Andrew N. and Hardison, Ross C. and Mackay, Joel P. and Blobel, Gerd A.},\\n doi = {10.1073/pnas.1102140108},\\n journal = {Proceedings of the National Academy of Sciences},\\n number = {22}\\n}\",\"author_short\":[\"Lamonica,  J., M.\",\"Deng,  W.\",\"Kadauke,  S.\",\"Campbell,  A., E.\",\"Gamsjaeger,  R.\",\"Wang,  H.\",\"Cheng,  Y.\",\"Billin,  A., N.\",\"Hardison,  R., C.\",\"Mackay,  J., P.\",\"Blobel,  G., A.\"],\"urls\":{\"Website\":\"http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011\",\"role\":\"author\",\"keyword\":[\"Gene regulation\",\"Hematopoiesis\",\"Posttranslational modifications\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1\",\"type\":\"article\",\"year\":\"2014\",\"keywords\":\"Breast cancer,DEAF-1,Embryonic development,LMO4,Transcriptional complex\",\"pages\":\"141-144\",\"volume\":\"8\",\"websites\":\"http://link.springer.com/10.1007/s12104-013-9470-x\",\"month\":\"4\",\"day\":\"16\",\"id\":\"70636852-15f1-3af2-95ff-10b3cc12cad6\",\"created\":\"2020-12-17T05:29:57.300Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:57.300Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Joseph2014a\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 2\",\"private_publication\":false,\"abstract\":\"The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.\",\"bibtype\":\"article\",\"author\":\"Joseph, Soumya and Kwan, Ann H. Y. and Mackay, Joel P. and Cubeddu, Liza and Matthews, Jacqueline M.\",\"doi\":\"10.1007/s12104-013-9470-x\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1},\\n type = {article},\\n year = {2014},\\n keywords = {Breast cancer,DEAF-1,Embryonic development,LMO4,Transcriptional complex},\\n pages = {141-144},\\n volume = {8},\\n websites = {http://link.springer.com/10.1007/s12104-013-9470-x},\\n month = {4},\\n day = {16},\\n id = {70636852-15f1-3af2-95ff-10b3cc12cad6},\\n created = {2020-12-17T05:29:57.300Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:57.300Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Joseph2014a},\\n source_type = {ARTICLE},\\n notes = {cited By 2},\\n private_publication = {false},\\n abstract = {The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.},\\n bibtype = {article},\\n author = {Joseph, Soumya and Kwan, Ann H. Y. and Mackay, Joel P. and Cubeddu, Liza and Matthews, Jacqueline M.},\\n doi = {10.1007/s12104-013-9470-x},\\n journal = {Biomolecular NMR Assignments},\\n number = {1}\\n}\",\"author_short\":[\"Joseph,  S.\",\"Kwan,  A., H., Y.\",\"Mackay,  J., P.\",\"Cubeddu,  L.\",\"Matthews,  J., M.\"],\"urls\":{\"Website\":\"http://link.springer.com/10.1007/s12104-013-9470-x\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014\",\"role\":\"author\",\"keyword\":[\"Breast cancer\",\"DEAF-1\",\"Embryonic development\",\"LMO4\",\"Transcriptional complex\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding\",\"type\":\"article\",\"year\":\"2018\",\"keywords\":\"P2RY12,blood platelet disorder,human,inherited,platelet dysfunction\",\"pages\":\"44-53\",\"volume\":\"16\",\"websites\":\"http://doi.wiley.com/10.1111/jth.13900\",\"month\":\"1\",\"id\":\"6ba6e7ee-21d0-3efa-a162-8e61a57a2e8e\",\"created\":\"2020-12-17T05:29:57.411Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:57.411Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Mundell2018\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 1\",\"private_publication\":false,\"abstract\":\"Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.\",\"bibtype\":\"article\",\"author\":\"Mundell, S. J. and Rabbolini, D. and Gabrielli, S. and Chen, Q. and Aungraheeta, R. and Hutchinson, J. L. and Kilo, T. and Mackay, J. and Ward, C. M. and Stevenson, W. and Morel-Kopp, M.-C.\",\"doi\":\"10.1111/jth.13900\",\"journal\":\"Journal of Thrombosis and Haemostasis\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding},\\n type = {article},\\n year = {2018},\\n keywords = {P2RY12,blood platelet disorder,human,inherited,platelet dysfunction},\\n pages = {44-53},\\n volume = {16},\\n websites = {http://doi.wiley.com/10.1111/jth.13900},\\n month = {1},\\n id = {6ba6e7ee-21d0-3efa-a162-8e61a57a2e8e},\\n created = {2020-12-17T05:29:57.411Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:57.411Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Mundell2018},\\n source_type = {ARTICLE},\\n notes = {cited By 1},\\n private_publication = {false},\\n abstract = {Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.},\\n bibtype = {article},\\n author = {Mundell, S. J. and Rabbolini, D. and Gabrielli, S. and Chen, Q. and Aungraheeta, R. and Hutchinson, J. L. and Kilo, T. and Mackay, J. and Ward, C. M. and Stevenson, W. and Morel-Kopp, M.-C.},\\n doi = {10.1111/jth.13900},\\n journal = {Journal of Thrombosis and Haemostasis},\\n number = {1}\\n}\",\"author_short\":[\"Mundell,  S., J.\",\"Rabbolini,  D.\",\"Gabrielli,  S.\",\"Chen,  Q.\",\"Aungraheeta,  R.\",\"Hutchinson,  J., L.\",\"Kilo,  T.\",\"Mackay,  J.\",\"Ward,  C., M.\",\"Stevenson,  W.\",\"Morel-Kopp,  M.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1111/jth.13900\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018\",\"role\":\"author\",\"keyword\":[\"P2RY12\",\"blood platelet disorder\",\"human\",\"inherited\",\"platelet dysfunction\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR\",\"type\":\"article\",\"year\":\"1997\",\"keywords\":\"Macromolecules,Pulsed-field-gradient NMR,Self-association,Solvent suppression,Translational diffusion coefficient\",\"pages\":\"1-8\",\"volume\":\"10\",\"websites\":\"https://link.springer.com/article/10.1023/A:1018339526108\",\"id\":\"c4559b9b-27f6-3507-a99a-884b885fa4fb\",\"created\":\"2020-12-17T05:29:57.841Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:57.841Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Dingley1997\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 24\",\"private_publication\":false,\"abstract\":\"We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.\",\"bibtype\":\"article\",\"author\":\"Dingley, Andrew J. and Mackay, Joel P. and Shaw, Graeme L. and Hambly, Brett D. and King, Glenn F.\",\"doi\":\"https://doi.org/10.1023/A:1018339526108\",\"journal\":\"Journal of Biomolecular NMR\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR},\\n type = {article},\\n year = {1997},\\n keywords = {Macromolecules,Pulsed-field-gradient NMR,Self-association,Solvent suppression,Translational diffusion coefficient},\\n pages = {1-8},\\n volume = {10},\\n websites = {https://link.springer.com/article/10.1023/A:1018339526108},\\n id = {c4559b9b-27f6-3507-a99a-884b885fa4fb},\\n created = {2020-12-17T05:29:57.841Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:57.841Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Dingley1997},\\n source_type = {ARTICLE},\\n notes = {cited By 24},\\n private_publication = {false},\\n abstract = {We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.},\\n bibtype = {article},\\n author = {Dingley, Andrew J. and Mackay, Joel P. and Shaw, Graeme L. and Hambly, Brett D. and King, Glenn F.},\\n doi = {https://doi.org/10.1023/A:1018339526108},\\n journal = {Journal of Biomolecular NMR},\\n number = {1}\\n}\",\"author_short\":[\"Dingley,  A., J.\",\"Mackay,  J., P.\",\"Shaw,  G., L.\",\"Hambly,  B., D.\",\"King,  G., F.\"],\"urls\":{\"Website\":\"https://link.springer.com/article/10.1023/A:1018339526108\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997\",\"role\":\"author\",\"keyword\":[\"Macromolecules\",\"Pulsed-field-gradient NMR\",\"Self-association\",\"Solvent suppression\",\"Translational diffusion coefficient\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G\",\"type\":\"article\",\"year\":\"2019\",\"keywords\":\"IPR020489,NMR,PF13989,RNA recognition motif,aminoglycoside,elongation factor\",\"pages\":\"699-705\",\"volume\":\"87\",\"websites\":\"http://doi.org/10.1002/prot.25687\",\"month\":\"8\",\"day\":\"22\",\"id\":\"7fb760d5-9bf0-3ae6-b357-d8046463887d\",\"created\":\"2020-12-17T05:29:57.892Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:57.892Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Mohanty2019\",\"source_type\":\"article\",\"private_publication\":false,\"abstract\":\"InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.\",\"bibtype\":\"article\",\"author\":\"Mohanty, Biswaranjan and Hanson‐Manful, Paulina and Finn, Thomas J. and Chambers, Cecilia R. and McKellar, James L. O. and Macindoe, Ingrid and Helder, Stephanie and Setiyaputra, Surya and Zhong, Yichen and Mackay, Joel P. and Patrick, Wayne M.\",\"doi\":\"10.1002/prot.25687\",\"journal\":\"Proteins: Structure, Function, and Bioinformatics\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G},\\n type = {article},\\n year = {2019},\\n keywords = {IPR020489,NMR,PF13989,RNA recognition motif,aminoglycoside,elongation factor},\\n pages = {699-705},\\n volume = {87},\\n websites = {http://doi.org/10.1002/prot.25687},\\n month = {8},\\n day = {22},\\n id = {7fb760d5-9bf0-3ae6-b357-d8046463887d},\\n created = {2020-12-17T05:29:57.892Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:57.892Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Mohanty2019},\\n source_type = {article},\\n private_publication = {false},\\n abstract = {InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.},\\n bibtype = {article},\\n author = {Mohanty, Biswaranjan and Hanson‐Manful, Paulina and Finn, Thomas J. and Chambers, Cecilia R. and McKellar, James L. O. and Macindoe, Ingrid and Helder, Stephanie and Setiyaputra, Surya and Zhong, Yichen and Mackay, Joel P. and Patrick, Wayne M.},\\n doi = {10.1002/prot.25687},\\n journal = {Proteins: Structure, Function, and Bioinformatics},\\n number = {8}\\n}\",\"author_short\":[\"Mohanty,  B.\",\"Hanson‐Manful,  P.\",\"Finn,  T., J.\",\"Chambers,  C., R.\",\"McKellar,  J., L., O.\",\"Macindoe,  I.\",\"Helder,  S.\",\"Setiyaputra,  S.\",\"Zhong,  Y.\",\"Mackay,  J., P.\",\"Patrick,  W., M.\"],\"urls\":{\"Website\":\"http://doi.org/10.1002/prot.25687\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019\",\"role\":\"author\",\"keyword\":[\"IPR020489\",\"NMR\",\"PF13989\",\"RNA recognition motif\",\"aminoglycoside\",\"elongation factor\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5\",\"type\":\"article\",\"year\":\"2014\",\"keywords\":\"Expression and purification,KOW domain,Spt4/5,Ubiquitin tag,pHUE\",\"pages\":\"54-60\",\"volume\":\"100\",\"websites\":\"https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989\",\"month\":\"8\",\"id\":\"c8758ecd-f201-3acf-9f44-56e1000a4f6a\",\"created\":\"2020-12-17T05:29:57.964Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:57.964Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Blythe2014\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 1\",\"private_publication\":false,\"abstract\":\"Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Blythe, Amanda and Gunasekara, Sanjika and Walshe, James and Mackay, Joel P. and Hartzog, Grant A. and Vrielink, Alice\",\"doi\":\"10.1016/j.pep.2014.05.005\",\"journal\":\"Protein Expression and Purification\",\"bibtex\":\"@article{\\n title = {Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5},\\n type = {article},\\n year = {2014},\\n keywords = {Expression and purification,KOW domain,Spt4/5,Ubiquitin tag,pHUE},\\n pages = {54-60},\\n volume = {100},\\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989},\\n month = {8},\\n id = {c8758ecd-f201-3acf-9f44-56e1000a4f6a},\\n created = {2020-12-17T05:29:57.964Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:57.964Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Blythe2014},\\n source_type = {ARTICLE},\\n notes = {cited By 1},\\n private_publication = {false},\\n abstract = {Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.},\\n bibtype = {article},\\n author = {Blythe, Amanda and Gunasekara, Sanjika and Walshe, James and Mackay, Joel P. and Hartzog, Grant A. and Vrielink, Alice},\\n doi = {10.1016/j.pep.2014.05.005},\\n journal = {Protein Expression and Purification}\\n}\",\"author_short\":[\"Blythe,  A.\",\"Gunasekara,  S.\",\"Walshe,  J.\",\"Mackay,  J., P.\",\"Hartzog,  G., A.\",\"Vrielink,  A.\"],\"urls\":{\"Website\":\"https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014\",\"role\":\"author\",\"keyword\":[\"Expression and purification\",\"KOW domain\",\"Spt4/5\",\"Ubiquitin tag\",\"pHUE\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Toward an estimation of binding constants in aqueous solution: studies of associations of vancomycin group antibiotics.\",\"type\":\"article\",\"year\":\"1993\",\"keywords\":\"Amide-amide hydrogen bond strengths,Enthalpic barriers,Enthalpy,Entropy compensation,Hydrophobic effect,Rotor restrictions\",\"pages\":\"1172-1178\",\"volume\":\"90\",\"websites\":\"http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172\",\"month\":\"2\",\"day\":\"15\",\"id\":\"5ef1cb1b-6f04-35c7-955d-7c5ca2d128fe\",\"created\":\"2020-12-17T05:29:58.125Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:58.125Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Williams1993\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 160\",\"private_publication\":false,\"abstract\":\"An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.\",\"bibtype\":\"article\",\"author\":\"Williams, Dudley H. and Searle, Mark S. and Mackay, Joel P. and Gerhard, Ute and Maplestone, Rachel A.\",\"doi\":\"10.1073/pnas.90.4.1172\",\"journal\":\"Proceedings of the National Academy of Sciences\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {Toward an estimation of binding constants in aqueous solution: studies of associations of vancomycin group antibiotics.},\\n type = {article},\\n year = {1993},\\n keywords = {Amide-amide hydrogen bond strengths,Enthalpic barriers,Enthalpy,Entropy compensation,Hydrophobic effect,Rotor restrictions},\\n pages = {1172-1178},\\n volume = {90},\\n websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172},\\n month = {2},\\n day = {15},\\n id = {5ef1cb1b-6f04-35c7-955d-7c5ca2d128fe},\\n created = {2020-12-17T05:29:58.125Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:58.125Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Williams1993},\\n source_type = {ARTICLE},\\n notes = {cited By 160},\\n private_publication = {false},\\n abstract = {An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.},\\n bibtype = {article},\\n author = {Williams, Dudley H. and Searle, Mark S. and Mackay, Joel P. and Gerhard, Ute and Maplestone, Rachel A.},\\n doi = {10.1073/pnas.90.4.1172},\\n journal = {Proceedings of the National Academy of Sciences},\\n number = {4}\\n}\",\"author_short\":[\"Williams,  D., H.\",\"Searle,  M., S.\",\"Mackay,  J., P.\",\"Gerhard,  U.\",\"Maplestone,  R., A.\"],\"urls\":{\"Website\":\"http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993\",\"role\":\"author\",\"keyword\":[\"Amide-amide hydrogen bond strengths\",\"Enthalpic barriers\",\"Enthalpy\",\"Entropy compensation\",\"Hydrophobic effect\",\"Rotor restrictions\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"type\":\"inbook\",\"year\":\"2018\",\"keywords\":\"Chemical exchange,Chemical shift perturbation,Cross-saturation,Dark states,Macromolecular NMR spectroscopy,Methyl-TROSY,Protein complexes,Protein-protein interactions\",\"pages\":\"2099-2132\",\"websites\":\"http://link.springer.com/10.1007/978-3-319-28388-3_121\",\"publisher\":\"Springer International Publishing\",\"city\":\"Cham\",\"id\":\"49e6d065-5bfa-3734-ac4c-302065de5fd0\",\"created\":\"2020-12-17T05:29:58.281Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:58.281Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Gell2018\",\"source_type\":\"article\",\"private_publication\":false,\"abstract\":\"Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.\",\"bibtype\":\"inbook\",\"author\":\"Gell, David A. and Kwan, Ann H. and Mackay, Joel P.\",\"doi\":\"10.1007/978-3-319-28388-3_121\",\"chapter\":\"NMR Spectroscopy in the Analysis of Protein-Protein Interactions\",\"title\":\"Modern Magnetic Resonance\",\"bibtex\":\"@inbook{\\n type = {inbook},\\n year = {2018},\\n keywords = {Chemical exchange,Chemical shift perturbation,Cross-saturation,Dark states,Macromolecular NMR spectroscopy,Methyl-TROSY,Protein complexes,Protein-protein interactions},\\n pages = {2099-2132},\\n websites = {http://link.springer.com/10.1007/978-3-319-28388-3_121},\\n publisher = {Springer International Publishing},\\n city = {Cham},\\n id = {49e6d065-5bfa-3734-ac4c-302065de5fd0},\\n created = {2020-12-17T05:29:58.281Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:58.281Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Gell2018},\\n source_type = {article},\\n private_publication = {false},\\n abstract = {Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.},\\n bibtype = {inbook},\\n author = {Gell, David A. and Kwan, Ann H. and Mackay, Joel P.},\\n doi = {10.1007/978-3-319-28388-3_121},\\n chapter = {NMR Spectroscopy in the Analysis of Protein-Protein Interactions},\\n title = {Modern Magnetic Resonance}\\n}\",\"author_short\":[\"Gell,  D., A.\",\"Kwan,  A., H.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://link.springer.com/10.1007/978-3-319-28388-3_121\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gell-kwan-mackay-modernmagneticresonance-2018\",\"role\":\"author\",\"keyword\":[\"Chemical exchange\",\"Chemical shift perturbation\",\"Cross-saturation\",\"Dark states\",\"Macromolecular NMR spectroscopy\",\"Methyl-TROSY\",\"Protein complexes\",\"Protein-protein interactions\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution Structures of Two CCHC Zinc Fingers from the FOG Family Protein U-Shaped that Mediate Protein–Protein Interactions\",\"type\":\"article\",\"year\":\"2000\",\"keywords\":\"GATA-1,Structure,Transcription,U-shaped,Zinc fingers\",\"pages\":\"1157-1166\",\"volume\":\"8\",\"websites\":\"https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X\",\"month\":\"11\",\"id\":\"ee4bb2bf-b119-390f-a5c0-747836832737\",\"created\":\"2020-12-17T05:29:58.497Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:58.497Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Liew2000\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 34\",\"private_publication\":false,\"abstract\":\"Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.\",\"bibtype\":\"article\",\"author\":\"Liew, Chu Kong and Kowalski, Kasper and Fox, Archa H. and Newton, Anthea and Sharpe, Belinda K. and Crossley, Merlin and Mackay, Joel P.\",\"doi\":\"10.1016/S0969-2126(00)00527-X\",\"journal\":\"Structure\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Solution Structures of Two CCHC Zinc Fingers from the FOG Family Protein U-Shaped that Mediate Protein–Protein Interactions},\\n type = {article},\\n year = {2000},\\n keywords = {GATA-1,Structure,Transcription,U-shaped,Zinc fingers},\\n pages = {1157-1166},\\n volume = {8},\\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X},\\n month = {11},\\n id = {ee4bb2bf-b119-390f-a5c0-747836832737},\\n created = {2020-12-17T05:29:58.497Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:58.497Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Liew2000},\\n source_type = {ARTICLE},\\n notes = {cited By 34},\\n private_publication = {false},\\n abstract = {Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.},\\n bibtype = {article},\\n author = {Liew, Chu Kong and Kowalski, Kasper and Fox, Archa H. and Newton, Anthea and Sharpe, Belinda K. and Crossley, Merlin and Mackay, Joel P.},\\n doi = {10.1016/S0969-2126(00)00527-X},\\n journal = {Structure},\\n number = {11}\\n}\",\"author_short\":[\"Liew,  C., K.\",\"Kowalski,  K.\",\"Fox,  A., H.\",\"Newton,  A.\",\"Sharpe,  B., K.\",\"Crossley,  M.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"liew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000\",\"role\":\"author\",\"keyword\":[\"GATA-1\",\"Structure\",\"Transcription\",\"U-shaped\",\"Zinc fingers\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural basis for rodlet assembly in fungal hydrophobins\",\"type\":\"article\",\"year\":\"2006\",\"keywords\":\"Amyloid,NMR,Polymer\",\"pages\":\"3621-3626\",\"volume\":\"103\",\"websites\":\"http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103\",\"month\":\"3\",\"day\":\"7\",\"id\":\"5c3390e3-fea1-3491-b3cb-82c763c2d1b5\",\"created\":\"2020-12-17T05:29:58.566Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:58.566Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Kwan2006\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 150\",\"private_publication\":false,\"abstract\":\"Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.\",\"bibtype\":\"article\",\"author\":\"Kwan, A. H. Y. and Winefield, R. D. and Sunde, M. and Matthews, J. M. and Haverkamp, R. G. and Templeton, M. D. and Mackay, J. P.\",\"doi\":\"10.1073/pnas.0505704103\",\"journal\":\"Proceedings of the National Academy of Sciences\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {Structural basis for rodlet assembly in fungal hydrophobins},\\n type = {article},\\n year = {2006},\\n keywords = {Amyloid,NMR,Polymer},\\n pages = {3621-3626},\\n volume = {103},\\n websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103},\\n month = {3},\\n day = {7},\\n id = {5c3390e3-fea1-3491-b3cb-82c763c2d1b5},\\n created = {2020-12-17T05:29:58.566Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:58.566Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Kwan2006},\\n source_type = {ARTICLE},\\n notes = {cited By 150},\\n private_publication = {false},\\n abstract = {Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.},\\n bibtype = {article},\\n author = {Kwan, A. H. Y. and Winefield, R. D. and Sunde, M. and Matthews, J. M. and Haverkamp, R. G. and Templeton, M. D. and Mackay, J. P.},\\n doi = {10.1073/pnas.0505704103},\\n journal = {Proceedings of the National Academy of Sciences},\\n number = {10}\\n}\",\"author_short\":[\"Kwan,  A., H., Y.\",\"Winefield,  R., D.\",\"Sunde,  M.\",\"Matthews,  J., M.\",\"Haverkamp,  R., G.\",\"Templeton,  M., D.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006\",\"role\":\"author\",\"keyword\":[\"Amyloid\",\"NMR\",\"Polymer\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP\",\"type\":\"article\",\"year\":\"2006\",\"keywords\":\"NMR,haem,haem-binding protein,p22HBP,protein structure\",\"pages\":\"287-297\",\"volume\":\"362\",\"websites\":\"https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679\",\"month\":\"9\",\"id\":\"6eee0455-9c97-35b4-ad59-54f7f318abd4\",\"created\":\"2020-12-17T05:29:58.858Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:58.858Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Gell2006\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 6\",\"private_publication\":false,\"abstract\":\"The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Gell, David A. and Westman, Belinda J. and Gorman, Daniel and Liew, ChuKong and Welch, John J. and Weiss, Mitchell J. and Mackay, Joel P.\",\"doi\":\"10.1016/j.jmb.2006.07.010\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP},\\n type = {article},\\n year = {2006},\\n keywords = {NMR,haem,haem-binding protein,p22HBP,protein structure},\\n pages = {287-297},\\n volume = {362},\\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679},\\n month = {9},\\n id = {6eee0455-9c97-35b4-ad59-54f7f318abd4},\\n created = {2020-12-17T05:29:58.858Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:58.858Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Gell2006},\\n source_type = {ARTICLE},\\n notes = {cited By 6},\\n private_publication = {false},\\n abstract = {The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Gell, David A. and Westman, Belinda J. and Gorman, Daniel and Liew, ChuKong and Welch, John J. and Weiss, Mitchell J. and Mackay, Joel P.},\\n doi = {10.1016/j.jmb.2006.07.010},\\n journal = {Journal of Molecular Biology},\\n number = {2}\\n}\",\"author_short\":[\"Gell,  D., A.\",\"Westman,  B., J.\",\"Gorman,  D.\",\"Liew,  C.\",\"Welch,  J., J.\",\"Weiss,  M., J.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006\",\"role\":\"author\",\"keyword\":[\"NMR\",\"haem\",\"haem-binding protein\",\"p22HBP\",\"protein structure\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"GATA-1: One protein, many partners\",\"type\":\"article\",\"year\":\"2006\",\"keywords\":\"Factor,Finger,Hematopoiesis,Transcription,Zinc\",\"pages\":\"6-11\",\"volume\":\"38\",\"websites\":\"https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074\",\"month\":\"1\",\"id\":\"ef48217b-ded1-36e7-b8a7-8426ac4c459e\",\"created\":\"2020-12-17T05:29:58.983Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:58.983Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Lowry2006\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 35\",\"private_publication\":false,\"abstract\":\"GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Lowry, Jason A. and Mackay, Joel P.\",\"doi\":\"10.1016/j.biocel.2005.06.017\",\"journal\":\"The International Journal of Biochemistry & Cell Biology\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {GATA-1: One protein, many partners},\\n type = {article},\\n year = {2006},\\n keywords = {Factor,Finger,Hematopoiesis,Transcription,Zinc},\\n pages = {6-11},\\n volume = {38},\\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074},\\n month = {1},\\n id = {ef48217b-ded1-36e7-b8a7-8426ac4c459e},\\n created = {2020-12-17T05:29:58.983Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:58.983Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Lowry2006},\\n source_type = {ARTICLE},\\n notes = {cited By 35},\\n private_publication = {false},\\n abstract = {GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Lowry, Jason A. and Mackay, Joel P.},\\n doi = {10.1016/j.biocel.2005.06.017},\\n journal = {The International Journal of Biochemistry & Cell Biology},\\n number = {1}\\n}\",\"author_short\":[\"Lowry,  J., A.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"lowry-mackay-gata1oneproteinmanypartners-2006\",\"role\":\"author\",\"keyword\":[\"Factor\",\"Finger\",\"Hematopoiesis\",\"Transcription\",\"Zinc\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7\",\"type\":\"article\",\"year\":\"2016\",\"keywords\":\"MTA1,NuRD complex,RBBP4,chromatin,protein structure,transcription regulation\",\"pages\":\"1472-1482\",\"volume\":\"25\",\"websites\":\"http://doi.wiley.com/10.1002/pro.2943\",\"month\":\"8\",\"id\":\"793f3c6b-8404-3b6e-9502-bf9b0357d1ca\",\"created\":\"2020-12-17T05:29:59.209Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:59.209Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Schmidberger2016\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 4\",\"private_publication\":false,\"abstract\":\"The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.\",\"bibtype\":\"article\",\"author\":\"Schmidberger, Jason W. and Sharifi Tabar, Mehdi and Torrado, Mario and Silva, Ana P. G. and Landsberg, Michael J. and Brillault, Lou and AlQarni, Saad and Zeng, Yi Cheng and Parker, Benjamin L. and Low, Jason K. K. and Mackay, Joel P.\",\"doi\":\"10.1002/pro.2943\",\"journal\":\"Protein Science\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7},\\n type = {article},\\n year = {2016},\\n keywords = {MTA1,NuRD complex,RBBP4,chromatin,protein structure,transcription regulation},\\n pages = {1472-1482},\\n volume = {25},\\n websites = {http://doi.wiley.com/10.1002/pro.2943},\\n month = {8},\\n id = {793f3c6b-8404-3b6e-9502-bf9b0357d1ca},\\n created = {2020-12-17T05:29:59.209Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:59.209Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Schmidberger2016},\\n source_type = {ARTICLE},\\n notes = {cited By 4},\\n private_publication = {false},\\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.},\\n bibtype = {article},\\n author = {Schmidberger, Jason W. and Sharifi Tabar, Mehdi and Torrado, Mario and Silva, Ana P. G. and Landsberg, Michael J. and Brillault, Lou and AlQarni, Saad and Zeng, Yi Cheng and Parker, Benjamin L. and Low, Jason K. K. and Mackay, Joel P.},\\n doi = {10.1002/pro.2943},\\n journal = {Protein Science},\\n number = {8}\\n}\",\"author_short\":[\"Schmidberger,  J., W.\",\"Sharifi Tabar,  M.\",\"Torrado,  M.\",\"Silva,  A., P., G.\",\"Landsberg,  M., J.\",\"Brillault,  L.\",\"AlQarni,  S.\",\"Zeng,  Y., C.\",\"Parker,  B., L.\",\"Low,  J., K., K.\",\"Mackay,  J., P.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1002/pro.2943\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016\",\"role\":\"author\",\"keyword\":[\"MTA1\",\"NuRD complex\",\"RBBP4\",\"chromatin\",\"protein structure\",\"transcription regulation\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Two-state conformational equilibrium in the Par-4 leucine zipper domain\",\"type\":\"article\",\"year\":\"2010\",\"keywords\":\"Circular dichroism,Leucine zipper,Prostate apoptosis response factor 4,Solution NMR spectroscopy\",\"pages\":\"n/a-n/a\",\"volume\":\"78\",\"websites\":\"http://doi.wiley.com/10.1002/prot.22752\",\"month\":\"4\",\"day\":\"15\",\"id\":\"90ab4173-c600-3c15-be8a-4dcbdfc4ed4d\",\"created\":\"2020-12-17T05:29:59.280Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T05:29:59.280Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":\"true\",\"hidden\":false,\"citation_key\":\"Schwalbe2010\",\"source_type\":\"ARTICLE\",\"notes\":\"cited By 5\",\"private_publication\":false,\"abstract\":\"Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.\",\"bibtype\":\"article\",\"author\":\"Schwalbe, Martin and Dutta, Kaushik and Libich, David S. and Venugopal, Hariprasad and Claridge, Jolyon K. and Gell, David A. and Mackay, Joel P. and Edwards, Patrick J. B. and Pascal, Steven M.\",\"doi\":\"10.1002/prot.22752\",\"journal\":\"Proteins: Structure, Function, and Bioinformatics\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Two-state conformational equilibrium in the Par-4 leucine zipper domain},\\n type = {article},\\n year = {2010},\\n keywords = {Circular dichroism,Leucine zipper,Prostate apoptosis response factor 4,Solution NMR spectroscopy},\\n pages = {n/a-n/a},\\n volume = {78},\\n websites = {http://doi.wiley.com/10.1002/prot.22752},\\n month = {4},\\n day = {15},\\n id = {90ab4173-c600-3c15-be8a-4dcbdfc4ed4d},\\n created = {2020-12-17T05:29:59.280Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T05:29:59.280Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {true},\\n hidden = {false},\\n citation_key = {Schwalbe2010},\\n source_type = {ARTICLE},\\n notes = {cited By 5},\\n private_publication = {false},\\n abstract = {Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.},\\n bibtype = {article},\\n author = {Schwalbe, Martin and Dutta, Kaushik and Libich, David S. and Venugopal, Hariprasad and Claridge, Jolyon K. and Gell, David A. and Mackay, Joel P. and Edwards, Patrick J. B. and Pascal, Steven M.},\\n doi = {10.1002/prot.22752},\\n journal = {Proteins: Structure, Function, and Bioinformatics},\\n number = {11}\\n}\",\"author_short\":[\"Schwalbe,  M.\",\"Dutta,  K.\",\"Libich,  D., S.\",\"Venugopal,  H.\",\"Claridge,  J., K.\",\"Gell,  D., A.\",\"Mackay,  J., P.\",\"Edwards,  P., J., B.\",\"Pascal,  S., M.\"],\"urls\":{\"Website\":\"http://doi.wiley.com/10.1002/prot.22752\"},\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010\",\"role\":\"author\",\"keyword\":[\"Circular dichroism\",\"Leucine zipper\",\"Prostate apoptosis response factor 4\",\"Solution NMR spectroscopy\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Cyclic peptides can engage a single binding pocket through highly divergent modes\",\"type\":\"article\",\"year\":\"2020\",\"keywords\":\"BET bromodomain inhibition,BRD3,BRD4,De novo cyclic peptides,Structural biology\",\"volume\":\"117\",\"id\":\"6636c6a0-e2f0-3aea-af8c-7865a5f17fcf\",\"created\":\"2020-12-17T07:26:41.435Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2020-12-17T07:26:41.435Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"© 2020 National Academy of Sciences. All rights reserved. Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.\",\"bibtype\":\"article\",\"author\":\"Patel, K. and Walport, L.J. and Walshe, J.L. and Solomon, P.D. and Low, J.K.K. and Tran, D.H. and Mouradian, K.S. and Silva, A.P.G. and Wilkinson-White, L. and Norman, A. and Franck, C. and Matthews, J.M. and Mitchell Guss, J. and Payne, R.J. and Passioura, T. and Suga, H. and Mackay, J.P.\",\"doi\":\"10.1073/pnas.2003086117\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"43\",\"bibtex\":\"@article{\\n title = {Cyclic peptides can engage a single binding pocket through highly divergent modes},\\n type = {article},\\n year = {2020},\\n keywords = {BET bromodomain inhibition,BRD3,BRD4,De novo cyclic peptides,Structural biology},\\n volume = {117},\\n id = {6636c6a0-e2f0-3aea-af8c-7865a5f17fcf},\\n created = {2020-12-17T07:26:41.435Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2020-12-17T07:26:41.435Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {© 2020 National Academy of Sciences. All rights reserved. Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.},\\n bibtype = {article},\\n author = {Patel, K. and Walport, L.J. and Walshe, J.L. and Solomon, P.D. and Low, J.K.K. and Tran, D.H. and Mouradian, K.S. and Silva, A.P.G. and Wilkinson-White, L. and Norman, A. and Franck, C. and Matthews, J.M. and Mitchell Guss, J. and Payne, R.J. and Passioura, T. and Suga, H. and Mackay, J.P.},\\n doi = {10.1073/pnas.2003086117},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {43}\\n}\",\"author_short\":[\"Patel,  K.\",\"Walport,  L.\",\"Walshe,  J.\",\"Solomon,  P.\",\"Low,  J.\",\"Tran,  D.\",\"Mouradian,  K.\",\"Silva,  A.\",\"Wilkinson-White,  L.\",\"Norman,  A.\",\"Franck,  C.\",\"Matthews,  J.\",\"Mitchell Guss,  J.\",\"Payne,  R.\",\"Passioura,  T.\",\"Suga,  H.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020\",\"role\":\"author\",\"urls\":{},\"keyword\":[\"BET bromodomain inhibition\",\"BRD3\",\"BRD4\",\"De novo cyclic peptides\",\"Structural biology\"],\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Site-selective photocatalytic functionalization of peptides and proteins at selenocysteine\",\"type\":\"article\",\"year\":\"2022\",\"volume\":\"13\",\"id\":\"659f1a34-764d-3452-8843-3347bb2c9170\",\"created\":\"2023-01-10T01:43:41.173Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:41.173Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The importance of modified peptides and proteins for applications in drug discovery, and for illuminating biological processes at the molecular level, is fueling a demand for efficient methods that facilitate the precise modification of these biomolecules. Herein, we describe the development of a photocatalytic method for the rapid and efficient dimerization and site-specific functionalization of peptide and protein diselenides. This methodology, dubbed the photocatalytic diselenide contraction, involves irradiation at 450 nm in the presence of an iridium photocatalyst and a phosphine and results in rapid and clean conversion of diselenides to reductively stable selenoethers. A mechanism for this photocatalytic transformation is proposed, which is supported by photoluminescence spectroscopy and density functional theory calculations. The utility of the photocatalytic diselenide contraction transformation is highlighted through the dimerization of selenopeptides, and by the generation of two families of protein conjugates via the site-selective modification of calmodulin containing the 21st amino acid selenocysteine, and the C-terminal modification of a ubiquitin diselenide.\",\"bibtype\":\"article\",\"author\":\"Dowman, L.J. and Kulkarni, S.S. and Alegre-Requena, J.V. and Giltrap, A.M. and Norman, A.R. and Sharma, A. and Gallegos, L.C. and Mackay, A.S. and Welegedara, A.P. and Watson, E.E. and Paton, R.S. and Payne, R.J.\",\"doi\":\"10.1038/s41467-022-34530-z\",\"journal\":\"Nature Communications\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Site-selective photocatalytic functionalization of peptides and proteins at selenocysteine},\\n type = {article},\\n year = {2022},\\n volume = {13},\\n id = {659f1a34-764d-3452-8843-3347bb2c9170},\\n created = {2023-01-10T01:43:41.173Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:41.173Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The importance of modified peptides and proteins for applications in drug discovery, and for illuminating biological processes at the molecular level, is fueling a demand for efficient methods that facilitate the precise modification of these biomolecules. Herein, we describe the development of a photocatalytic method for the rapid and efficient dimerization and site-specific functionalization of peptide and protein diselenides. This methodology, dubbed the photocatalytic diselenide contraction, involves irradiation at 450 nm in the presence of an iridium photocatalyst and a phosphine and results in rapid and clean conversion of diselenides to reductively stable selenoethers. A mechanism for this photocatalytic transformation is proposed, which is supported by photoluminescence spectroscopy and density functional theory calculations. The utility of the photocatalytic diselenide contraction transformation is highlighted through the dimerization of selenopeptides, and by the generation of two families of protein conjugates via the site-selective modification of calmodulin containing the 21st amino acid selenocysteine, and the C-terminal modification of a ubiquitin diselenide.},\\n bibtype = {article},\\n author = {Dowman, L.J. and Kulkarni, S.S. and Alegre-Requena, J.V. and Giltrap, A.M. and Norman, A.R. and Sharma, A. and Gallegos, L.C. and Mackay, A.S. and Welegedara, A.P. and Watson, E.E. and Paton, R.S. and Payne, R.J.},\\n doi = {10.1038/s41467-022-34530-z},\\n journal = {Nature Communications},\\n number = {1}\\n}\",\"author_short\":[\"Dowman,  L.\",\"Kulkarni,  S.\",\"Alegre-Requena,  J.\",\"Giltrap,  A.\",\"Norman,  A.\",\"Sharma,  A.\",\"Gallegos,  L.\",\"Mackay,  A.\",\"Welegedara,  A.\",\"Watson,  E.\",\"Paton,  R.\",\"Payne,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dowman-kulkarni-alegrerequena-giltrap-norman-sharma-gallegos-mackay-etal-siteselectivephotocatalyticfunctionalizationofpeptidesandproteinsatselenocysteine-2022\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The role of auxiliary domains in modulating CHD4 activity suggests mechanistic commonality between enzyme families\",\"type\":\"article\",\"year\":\"2022\",\"volume\":\"13\",\"id\":\"76602a66-c88f-3ef1-8f5b-ae0a77f5e423\",\"created\":\"2023-01-10T01:43:42.100Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:42.100Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"CHD4 is an essential, widely conserved ATP-dependent translocase that is also a broad tumour dependency. In common with other SF2-family chromatin remodelling enzymes, it alters chromatin accessibility by repositioning histone octamers. Besides the helicase and adjacent tandem chromodomains and PHD domains, CHD4 features 1000 residues of N- and C-terminal sequence with unknown structure and function. We demonstrate that these regions regulate CHD4 activity through different mechanisms. An N-terminal intrinsically disordered region (IDR) promotes remodelling integrity in a manner that depends on the composition but not sequence of the IDR. The C-terminal region harbours an auto-inhibitory region that contacts the helicase domain. Auto-inhibition is relieved by a previously unrecognized C-terminal SANT-SLIDE domain split by ~150 residues of disordered sequence, most likely by binding of this domain to substrate DNA. Our data shed light on CHD4 regulation and reveal strong mechanistic commonality between CHD family members, as well as with ISWI-family remodellers.\",\"bibtype\":\"article\",\"author\":\"Zhong, Y. and Moghaddas Sani, H. and Paudel, B.P. and Low, J.K.K. and Silva, A.P.G. and Mueller, S. and Deshpande, C. and Panjikar, S. and Reid, X.J. and Bedward, M.J. and van Oijen, A.M. and Mackay, J.P.\",\"doi\":\"10.1038/s41467-022-35002-0\",\"journal\":\"Nature Communications\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {The role of auxiliary domains in modulating CHD4 activity suggests mechanistic commonality between enzyme families},\\n type = {article},\\n year = {2022},\\n volume = {13},\\n id = {76602a66-c88f-3ef1-8f5b-ae0a77f5e423},\\n created = {2023-01-10T01:43:42.100Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:42.100Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {CHD4 is an essential, widely conserved ATP-dependent translocase that is also a broad tumour dependency. In common with other SF2-family chromatin remodelling enzymes, it alters chromatin accessibility by repositioning histone octamers. Besides the helicase and adjacent tandem chromodomains and PHD domains, CHD4 features 1000 residues of N- and C-terminal sequence with unknown structure and function. We demonstrate that these regions regulate CHD4 activity through different mechanisms. An N-terminal intrinsically disordered region (IDR) promotes remodelling integrity in a manner that depends on the composition but not sequence of the IDR. The C-terminal region harbours an auto-inhibitory region that contacts the helicase domain. Auto-inhibition is relieved by a previously unrecognized C-terminal SANT-SLIDE domain split by ~150 residues of disordered sequence, most likely by binding of this domain to substrate DNA. Our data shed light on CHD4 regulation and reveal strong mechanistic commonality between CHD family members, as well as with ISWI-family remodellers.},\\n bibtype = {article},\\n author = {Zhong, Y. and Moghaddas Sani, H. and Paudel, B.P. and Low, J.K.K. and Silva, A.P.G. and Mueller, S. and Deshpande, C. and Panjikar, S. and Reid, X.J. and Bedward, M.J. and van Oijen, A.M. and Mackay, J.P.},\\n doi = {10.1038/s41467-022-35002-0},\\n journal = {Nature Communications},\\n number = {1}\\n}\",\"author_short\":[\"Zhong,  Y.\",\"Moghaddas Sani,  H.\",\"Paudel,  B.\",\"Low,  J.\",\"Silva,  A.\",\"Mueller,  S.\",\"Deshpande,  C.\",\"Panjikar,  S.\",\"Reid,  X.\",\"Bedward,  M.\",\"van Oijen,  A.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"zhong-moghaddassani-paudel-low-silva-mueller-deshpande-panjikar-etal-theroleofauxiliarydomainsinmodulatingchd4activitysuggestsmechanisticcommonalitybetweenenzymefamilies-2022\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":4,\"html\":\"\"},{\"title\":\"Mucosal TLR2-activating protein-based vaccination induces potent pulmonary immunity and protection against SARS-CoV-2 in mice\",\"type\":\"article\",\"year\":\"2022\",\"volume\":\"13\",\"id\":\"817b23fa-ab01-305f-8861-84807b7e3327\",\"created\":\"2023-01-10T01:43:43.018Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:43.018Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Current vaccines against SARS-CoV-2 substantially reduce mortality, but protection against infection is less effective. Enhancing immunity in the respiratory tract, via mucosal vaccination, may provide protection against infection and minimise viral spread. Here, we report testing of a subunit vaccine in mice, consisting of SARS-CoV-2 Spike protein with a TLR2-stimulating adjuvant (Pam2Cys), delivered to mice parenterally or mucosally. Both routes of vaccination induce substantial neutralising antibody (nAb) titres, however, mucosal vaccination uniquely generates anti-Spike IgA, increases nAb in the serum and airways, and increases lung CD4+ T-cell responses. TLR2 is expressed by respiratory epithelia and immune cells. Using TLR2 deficient chimeric mice, we determine that TLR2 expression in either compartment facilitates early innate responses to mucosal vaccination. By contrast, TLR2 on hematopoietic cells is essential for optimal lung-localised, antigen-specific responses. In K18-hACE2 mice, vaccination provides complete protection against disease and sterilising lung immunity against SARS-CoV-2, with a short-term non-specific protective effect from mucosal Pam2Cys alone. These data support mucosal vaccination as a strategy to improve protection in the respiratory tract against SARS-CoV-2 and other respiratory viruses.\",\"bibtype\":\"article\",\"author\":\"Ashhurst, A.S. and Johansen, M.D. and Maxwell, J.W.C. and Stockdale, S. and Ashley, C.L. and Aggarwal, A. and Siddiquee, R. and Miemczyk, S. and Nguyen, D.H. and Mackay, J.P. and Payne, R.J. and Britton, W.J.\",\"doi\":\"10.1038/s41467-022-34297-3\",\"journal\":\"Nature Communications\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Mucosal TLR2-activating protein-based vaccination induces potent pulmonary immunity and protection against SARS-CoV-2 in mice},\\n type = {article},\\n year = {2022},\\n volume = {13},\\n id = {817b23fa-ab01-305f-8861-84807b7e3327},\\n created = {2023-01-10T01:43:43.018Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:43.018Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Current vaccines against SARS-CoV-2 substantially reduce mortality, but protection against infection is less effective. Enhancing immunity in the respiratory tract, via mucosal vaccination, may provide protection against infection and minimise viral spread. Here, we report testing of a subunit vaccine in mice, consisting of SARS-CoV-2 Spike protein with a TLR2-stimulating adjuvant (Pam2Cys), delivered to mice parenterally or mucosally. Both routes of vaccination induce substantial neutralising antibody (nAb) titres, however, mucosal vaccination uniquely generates anti-Spike IgA, increases nAb in the serum and airways, and increases lung CD4+ T-cell responses. TLR2 is expressed by respiratory epithelia and immune cells. Using TLR2 deficient chimeric mice, we determine that TLR2 expression in either compartment facilitates early innate responses to mucosal vaccination. By contrast, TLR2 on hematopoietic cells is essential for optimal lung-localised, antigen-specific responses. In K18-hACE2 mice, vaccination provides complete protection against disease and sterilising lung immunity against SARS-CoV-2, with a short-term non-specific protective effect from mucosal Pam2Cys alone. These data support mucosal vaccination as a strategy to improve protection in the respiratory tract against SARS-CoV-2 and other respiratory viruses.},\\n bibtype = {article},\\n author = {Ashhurst, A.S. and Johansen, M.D. and Maxwell, J.W.C. and Stockdale, S. and Ashley, C.L. and Aggarwal, A. and Siddiquee, R. and Miemczyk, S. and Nguyen, D.H. and Mackay, J.P. and Payne, R.J. and Britton, W.J.},\\n doi = {10.1038/s41467-022-34297-3},\\n journal = {Nature Communications},\\n number = {1}\\n}\",\"author_short\":[\"Ashhurst,  A.\",\"Johansen,  M.\",\"Maxwell,  J.\",\"Stockdale,  S.\",\"Ashley,  C.\",\"Aggarwal,  A.\",\"Siddiquee,  R.\",\"Miemczyk,  S.\",\"Nguyen,  D.\",\"Mackay,  J.\",\"Payne,  R.\",\"Britton,  W.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"ashhurst-johansen-maxwell-stockdale-ashley-aggarwal-siddiquee-miemczyk-etal-mucosaltlr2activatingproteinbasedvaccinationinducespotentpulmonaryimmunityandprotectionagainstsarscov2inmice-2022\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"RNA inhibits dMi-2/CHD4 chromatin binding and nucleosome remodeling\",\"type\":\"article\",\"year\":\"2022\",\"volume\":\"39\",\"id\":\"d99d4ee9-901c-3e7f-9a93-d47a33ffacef\",\"created\":\"2023-01-10T01:43:43.942Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:43.942Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The ATP-dependent nucleosome remodeler Mi-2/CHD4 broadly modulates chromatin landscapes to repress transcription and to maintain genome integrity. Here we use individual nucleotide resolution crosslinking and immunoprecipitation (iCLIP) to show that Drosophila Mi-2 associates with thousands of mRNA molecules in vivo. Biochemical data reveal that recombinant dMi-2 preferentially binds to G-rich RNA molecules using two intrinsically disordered regions of unclear function. Pharmacological inhibition of transcription and RNase digestion approaches establish that RNA inhibits the association of dMi-2 with chromatin. We also show that RNA inhibits dMi-2-mediated nucleosome mobilization by competing with the nucleosome substrate. Importantly, this activity is shared by CHD4, the human homolog of dMi-2, strongly suggesting that RNA-mediated regulation of remodeler activity is an evolutionary conserved mechanism. Our data support a model in which RNA serves to protect actively transcribed regions of the genome from dMi-2/CHD4-mediated establishment of repressive chromatin structures.\",\"bibtype\":\"article\",\"author\":\"Ullah, I. and Thölken, C. and Zhong, Y. and John, M. and Rossbach, O. and Lenz, J. and Gößringer, M. and Nist, A. and Albert, L. and Stiewe, T. and Mackay, J.P. and Brehm, A.\",\"doi\":\"10.1016/j.celrep.2022.110895\",\"journal\":\"Cell Reports\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {RNA inhibits dMi-2/CHD4 chromatin binding and nucleosome remodeling},\\n type = {article},\\n year = {2022},\\n volume = {39},\\n id = {d99d4ee9-901c-3e7f-9a93-d47a33ffacef},\\n created = {2023-01-10T01:43:43.942Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:43.942Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The ATP-dependent nucleosome remodeler Mi-2/CHD4 broadly modulates chromatin landscapes to repress transcription and to maintain genome integrity. Here we use individual nucleotide resolution crosslinking and immunoprecipitation (iCLIP) to show that Drosophila Mi-2 associates with thousands of mRNA molecules in vivo. Biochemical data reveal that recombinant dMi-2 preferentially binds to G-rich RNA molecules using two intrinsically disordered regions of unclear function. Pharmacological inhibition of transcription and RNase digestion approaches establish that RNA inhibits the association of dMi-2 with chromatin. We also show that RNA inhibits dMi-2-mediated nucleosome mobilization by competing with the nucleosome substrate. Importantly, this activity is shared by CHD4, the human homolog of dMi-2, strongly suggesting that RNA-mediated regulation of remodeler activity is an evolutionary conserved mechanism. Our data support a model in which RNA serves to protect actively transcribed regions of the genome from dMi-2/CHD4-mediated establishment of repressive chromatin structures.},\\n bibtype = {article},\\n author = {Ullah, I. and Thölken, C. and Zhong, Y. and John, M. and Rossbach, O. and Lenz, J. and Gößringer, M. and Nist, A. and Albert, L. and Stiewe, T. and Mackay, J.P. and Brehm, A.},\\n doi = {10.1016/j.celrep.2022.110895},\\n journal = {Cell Reports},\\n number = {9}\\n}\",\"author_short\":[\"Ullah,  I.\",\"Thölken,  C.\",\"Zhong,  Y.\",\"John,  M.\",\"Rossbach,  O.\",\"Lenz,  J.\",\"Gößringer,  M.\",\"Nist,  A.\",\"Albert,  L.\",\"Stiewe,  T.\",\"Mackay,  J.\",\"Brehm,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"ullah-thlken-zhong-john-rossbach-lenz-gringer-nist-etal-rnainhibitsdmi2chd4chromatinbindingandnucleosomeremodeling-2022\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Mechanism of Bloom syndrome complex assembly required for double Holliday junction dissolution and genome stability\",\"type\":\"article\",\"year\":\"2022\",\"volume\":\"119\",\"id\":\"eea6affc-fd8d-3936-9453-941dacf51a3d\",\"created\":\"2023-01-10T01:43:44.899Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:44.899Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The RecQ-like helicase BLM cooperates with topoisomerase IIIα, RMI1, and RMI2 in a heterotetrameric complex (the “Bloom syndrome complex”) for dissolution of double Holliday junctions, key intermediates in homologous recombination. Mutations in any component of the Bloom syndrome complex can cause genome instability and a highly cancer-prone disorder called Bloom syndrome. Some heterozygous carriers are also predisposed to breast cancer. To understand how the activities of BLM helicase and topoisomerase IIIα are coupled, we purified the active four-subunit complex. Chemical cross-linking and mass spectrometry revealed a unique architecture that links the helicase and topoisomerase domains. Using biochemical experiments, we demonstrated dimerization mediated by the N terminus of BLM with a 2:2:2:2 stoichiometry within the Bloom syndrome complex. We identified mutations that independently abrogate dimerization or association of BLM with RMI1, and we show that both are dysfunctional for dissolution using in vitro assays and cause genome instability and synthetic lethal interactions with GEN1/MUS81 in cells. Truncated BLM can also inhibit the activity of full-length BLM in mixed dimers, suggesting a putative mechanism of dominant-negative action in carriers of BLM truncation alleles. Our results identify critical molecular determinants of Bloom syndrome complex assembly required for double Holliday junction dissolution and maintenance of genome stability.\",\"bibtype\":\"article\",\"author\":\"Hodson, C. and Low, J.K.K. and van Twest, S. and Jones, S.E. and Swuec, P. and Murphy, V. and Tsukada, K. and Fawkes, M. and Bythell-Douglas, R. and Davies, A. and Costa, A. and Deans, A.J.\",\"doi\":\"10.1073/pnas.2109093119\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Mechanism of Bloom syndrome complex assembly required for double Holliday junction dissolution and genome stability},\\n type = {article},\\n year = {2022},\\n volume = {119},\\n id = {eea6affc-fd8d-3936-9453-941dacf51a3d},\\n created = {2023-01-10T01:43:44.899Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:44.899Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The RecQ-like helicase BLM cooperates with topoisomerase IIIα, RMI1, and RMI2 in a heterotetrameric complex (the “Bloom syndrome complex”) for dissolution of double Holliday junctions, key intermediates in homologous recombination. Mutations in any component of the Bloom syndrome complex can cause genome instability and a highly cancer-prone disorder called Bloom syndrome. Some heterozygous carriers are also predisposed to breast cancer. To understand how the activities of BLM helicase and topoisomerase IIIα are coupled, we purified the active four-subunit complex. Chemical cross-linking and mass spectrometry revealed a unique architecture that links the helicase and topoisomerase domains. Using biochemical experiments, we demonstrated dimerization mediated by the N terminus of BLM with a 2:2:2:2 stoichiometry within the Bloom syndrome complex. We identified mutations that independently abrogate dimerization or association of BLM with RMI1, and we show that both are dysfunctional for dissolution using in vitro assays and cause genome instability and synthetic lethal interactions with GEN1/MUS81 in cells. Truncated BLM can also inhibit the activity of full-length BLM in mixed dimers, suggesting a putative mechanism of dominant-negative action in carriers of BLM truncation alleles. Our results identify critical molecular determinants of Bloom syndrome complex assembly required for double Holliday junction dissolution and maintenance of genome stability.},\\n bibtype = {article},\\n author = {Hodson, C. and Low, J.K.K. and van Twest, S. and Jones, S.E. and Swuec, P. and Murphy, V. and Tsukada, K. and Fawkes, M. and Bythell-Douglas, R. and Davies, A. and Costa, A. and Deans, A.J.},\\n doi = {10.1073/pnas.2109093119},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {6}\\n}\",\"author_short\":[\"Hodson,  C.\",\"Low,  J.\",\"van Twest,  S.\",\"Jones,  S.\",\"Swuec,  P.\",\"Murphy,  V.\",\"Tsukada,  K.\",\"Fawkes,  M.\",\"Bythell-Douglas,  R.\",\"Davies,  A.\",\"Costa,  A.\",\"Deans,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"hodson-low-vantwest-jones-swuec-murphy-tsukada-fawkes-etal-mechanismofbloomsyndromecomplexassemblyrequiredfordoublehollidayjunctiondissolutionandgenomestability-2022\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Molecular architecture of nucleosome remodeling and deacetylase sub-complexes by integrative structure determination\",\"type\":\"article\",\"year\":\"2022\",\"volume\":\"31\",\"id\":\"eef4f63c-8daa-3623-b17b-a5cf28db9aee\",\"created\":\"2023-01-10T01:43:45.810Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:45.810Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The nucleosome remodeling and deacetylase (NuRD) complex is a chromatin-modifying assembly that regulates gene expression and DNA damage repair. Despite its importance, limited structural information describing the complete NuRD complex is available and a detailed understanding of its mechanism is therefore lacking. Drawing on information from SEC-MALLS, DIA-MS, XLMS, negative-stain EM, X-ray crystallography, NMR spectroscopy, secondary structure predictions, and homology models, we applied Bayesian integrative structure determination to investigate the molecular architecture of three NuRD sub-complexes: MTA1-HDAC1-RBBP4, MTA1N-HDAC1-MBD3GATAD2CC, and MTA1-HDAC1-RBBP4-MBD3-GATAD2A [nucleosome deacetylase (NuDe)]. The integrative structures were corroborated by examining independent crosslinks, cryo-EM maps, biochemical assays, known cancer-associated mutations, and structure predictions from AlphaFold. The robustness of the models was assessed by jack-knifing. Localization of the full-length MBD3, which connects the deacetylase and chromatin remodeling modules in NuRD, has not previously been possible; our models indicate two different locations for MBD3, suggesting a mechanism by which MBD3 in the presence of GATAD2A asymmetrically bridges the two modules in NuRD. Further, our models uncovered three previously unrecognized subunit interfaces in NuDe: HDAC1C-MTA1BAH, MTA1BAH-MBD3MBD, and HDAC160–100-MBD3MBD. Our approach also allowed us to localize regions of unknown structure, such as HDAC1C and MBD3IDR, thereby resulting in the most complete and robustly cross-validated structural characterization of these NuRD sub-complexes so far.\",\"bibtype\":\"article\",\"author\":\"Arvindekar, S. and Jackman, M.J. and Low, J.K.K. and Landsberg, M.J. and Mackay, J.P. and Viswanath, S.\",\"doi\":\"10.1002/pro.4387\",\"journal\":\"Protein Science\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {Molecular architecture of nucleosome remodeling and deacetylase sub-complexes by integrative structure determination},\\n type = {article},\\n year = {2022},\\n volume = {31},\\n id = {eef4f63c-8daa-3623-b17b-a5cf28db9aee},\\n created = {2023-01-10T01:43:45.810Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:45.810Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a chromatin-modifying assembly that regulates gene expression and DNA damage repair. Despite its importance, limited structural information describing the complete NuRD complex is available and a detailed understanding of its mechanism is therefore lacking. Drawing on information from SEC-MALLS, DIA-MS, XLMS, negative-stain EM, X-ray crystallography, NMR spectroscopy, secondary structure predictions, and homology models, we applied Bayesian integrative structure determination to investigate the molecular architecture of three NuRD sub-complexes: MTA1-HDAC1-RBBP4, MTA1N-HDAC1-MBD3GATAD2CC, and MTA1-HDAC1-RBBP4-MBD3-GATAD2A [nucleosome deacetylase (NuDe)]. The integrative structures were corroborated by examining independent crosslinks, cryo-EM maps, biochemical assays, known cancer-associated mutations, and structure predictions from AlphaFold. The robustness of the models was assessed by jack-knifing. Localization of the full-length MBD3, which connects the deacetylase and chromatin remodeling modules in NuRD, has not previously been possible; our models indicate two different locations for MBD3, suggesting a mechanism by which MBD3 in the presence of GATAD2A asymmetrically bridges the two modules in NuRD. Further, our models uncovered three previously unrecognized subunit interfaces in NuDe: HDAC1C-MTA1BAH, MTA1BAH-MBD3MBD, and HDAC160–100-MBD3MBD. Our approach also allowed us to localize regions of unknown structure, such as HDAC1C and MBD3IDR, thereby resulting in the most complete and robustly cross-validated structural characterization of these NuRD sub-complexes so far.},\\n bibtype = {article},\\n author = {Arvindekar, S. and Jackman, M.J. and Low, J.K.K. and Landsberg, M.J. and Mackay, J.P. and Viswanath, S.},\\n doi = {10.1002/pro.4387},\\n journal = {Protein Science},\\n number = {9}\\n}\",\"author_short\":[\"Arvindekar,  S.\",\"Jackman,  M.\",\"Low,  J.\",\"Landsberg,  M.\",\"Mackay,  J.\",\"Viswanath,  S.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"arvindekar-jackman-low-landsberg-mackay-viswanath-moleculararchitectureofnucleosomeremodelinganddeacetylasesubcomplexesbyintegrativestructuredetermination-2022\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"TNP Analogues Inhibit the Virulence Promoting IP<inf>3-4</inf> Kinase Arg1 in the Fungal Pathogen Cryptococcus neoformans\",\"type\":\"article\",\"year\":\"2022\",\"volume\":\"12\",\"id\":\"9cbc3fce-c9c1-35f8-ba6d-09bac2420d29\",\"created\":\"2023-01-10T01:43:46.835Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:46.835Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"New antifungals with unique modes of action are urgently needed to treat the increasing global burden of invasive fungal infections. The fungal inositol polyphosphate kinase (IPK) pathway, comprised of IPKs that convert IP3 to IP8, provides a promising new target due to its impact on multiple, critical cellular functions and, unlike in mammalian cells, its lack of redundancy. Nearly all IPKs in the fungal pathway are essential for virulence, with IP3-4 kinase (IP3-4K) the most critical. The dibenzylaminopurine compound, N2-(m-trifluorobenzylamino)-N6-(p-nitrobenzylamino)purine (TNP), is a commercially available inhibitor of mammalian IPKs. The ability of TNP to be adapted as an inhibitor of fungal IP3-4K has not been investigated. We purified IP3-4K from the human pathogens, Cryptococcus neoformans and Candida albicans, and optimised enzyme and surface plasmon resonance (SPR) assays to determine the half inhibitory concentration (IC50) and binding affinity (KD), respectively, of TNP and 38 analogues. A novel chemical route was developed to efficiently prepare TNP analogues. TNP and its analogues demonstrated inhibition of recombinant IP3-4K from C. neoformans (CnArg1) at low µM IC50s, but not IP3-4K from C. albicans (CaIpk2) and many analogues exhibited selectivity for CnArg1 over the human equivalent, HsIPMK. Our results provide a foundation for improving potency and selectivity of the TNP series for fungal IP3-4K.\",\"bibtype\":\"article\",\"author\":\"Desmarini, D. and Truong, D. and Wilkinson-White, L. and Desphande, C. and Torrado, M. and Mackay, J.P. and Matthews, J.M. and Sorrell, T.C. and Lev, S. and Thompson, P.E. and Thompson, P.E. and Djordjevic, J.T.\",\"doi\":\"10.3390/biom12101526\",\"journal\":\"Biomolecules\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {TNP Analogues Inhibit the Virulence Promoting IP<inf>3-4</inf> Kinase Arg1 in the Fungal Pathogen Cryptococcus neoformans},\\n type = {article},\\n year = {2022},\\n volume = {12},\\n id = {9cbc3fce-c9c1-35f8-ba6d-09bac2420d29},\\n created = {2023-01-10T01:43:46.835Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:46.835Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {New antifungals with unique modes of action are urgently needed to treat the increasing global burden of invasive fungal infections. The fungal inositol polyphosphate kinase (IPK) pathway, comprised of IPKs that convert IP3 to IP8, provides a promising new target due to its impact on multiple, critical cellular functions and, unlike in mammalian cells, its lack of redundancy. Nearly all IPKs in the fungal pathway are essential for virulence, with IP3-4 kinase (IP3-4K) the most critical. The dibenzylaminopurine compound, N2-(m-trifluorobenzylamino)-N6-(p-nitrobenzylamino)purine (TNP), is a commercially available inhibitor of mammalian IPKs. The ability of TNP to be adapted as an inhibitor of fungal IP3-4K has not been investigated. We purified IP3-4K from the human pathogens, Cryptococcus neoformans and Candida albicans, and optimised enzyme and surface plasmon resonance (SPR) assays to determine the half inhibitory concentration (IC50) and binding affinity (KD), respectively, of TNP and 38 analogues. A novel chemical route was developed to efficiently prepare TNP analogues. TNP and its analogues demonstrated inhibition of recombinant IP3-4K from C. neoformans (CnArg1) at low µM IC50s, but not IP3-4K from C. albicans (CaIpk2) and many analogues exhibited selectivity for CnArg1 over the human equivalent, HsIPMK. Our results provide a foundation for improving potency and selectivity of the TNP series for fungal IP3-4K.},\\n bibtype = {article},\\n author = {Desmarini, D. and Truong, D. and Wilkinson-White, L. and Desphande, C. and Torrado, M. and Mackay, J.P. and Matthews, J.M. and Sorrell, T.C. and Lev, S. and Thompson, P.E. and Thompson, P.E. and Djordjevic, J.T.},\\n doi = {10.3390/biom12101526},\\n journal = {Biomolecules},\\n number = {10}\\n}\",\"author_short\":[\"Desmarini,  D.\",\"Truong,  D.\",\"Wilkinson-White,  L.\",\"Desphande,  C.\",\"Torrado,  M.\",\"Mackay,  J.\",\"Matthews,  J.\",\"Sorrell,  T.\",\"Lev,  S.\",\"Thompson,  P.\",\"Thompson,  P.\",\"Djordjevic,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"desmarini-truong-wilkinsonwhite-desphande-torrado-mackay-matthews-sorrell-etal-tnpanaloguesinhibitthevirulencepromotingipinf34infkinasearg1inthefungalpathogencryptococcusneoformans-2022\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Unique protein interaction networks define the chromatin remodelling module of the NuRD complex\",\"type\":\"article\",\"year\":\"2022\",\"pages\":\"199-214\",\"volume\":\"289\",\"id\":\"a6268275-0bc8-3002-bce9-6d7a08deb3b1\",\"created\":\"2023-01-10T01:43:47.765Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:47.765Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The combination of four proteins and their paralogues including MBD2/3, GATAD2A/B, CDK2AP1 and CHD3/4/5, which we refer to as the MGCC module, form the chromatin remodelling module of the nucleosome remodelling and deacetylase (NuRD) complex. To date, mechanisms by which the MGCC module acquires paralogue-specific function and specificity have not been addressed. Understanding the protein–protein interaction (PPI) network of the MGCC subunits is essential for defining underlying mechanisms of gene regulation. Therefore, using pulldown followed by mass spectrometry analysis (PD-MS), we report a proteome-wide interaction network of the MGCC module in a paralogue-specific manner. Our data also demonstrate that the disordered C-terminal region of CHD3/4/5 is a gateway to incorporate remodelling activity into both ChAHP (CHD4, ADNP, HP1γ) and NuRD complexes in a mutually exclusive manner. We define a short aggregation-prone region (APR) within the C-terminal segment of GATAD2B that is essential for the interaction of CHD4 and CDK2AP1 with the NuRD complex. Finally, we also report an association of CDK2AP1 with the nuclear receptor co-repressor (NCOR) complex. Overall, this study provides insight into the possible mechanisms through which the MGCC module can achieve specificity and diverse biological functions.\",\"bibtype\":\"article\",\"author\":\"Sharifi Tabar, M. and Giardina, C. and Feng, Y. and Francis, H. and Moghaddas Sani, H. and Low, J.K.K. and Mackay, J.P. and Bailey, C.G. and Rasko, J.E.J.\",\"doi\":\"10.1111/febs.16112\",\"journal\":\"FEBS Journal\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Unique protein interaction networks define the chromatin remodelling module of the NuRD complex},\\n type = {article},\\n year = {2022},\\n pages = {199-214},\\n volume = {289},\\n id = {a6268275-0bc8-3002-bce9-6d7a08deb3b1},\\n created = {2023-01-10T01:43:47.765Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:47.765Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The combination of four proteins and their paralogues including MBD2/3, GATAD2A/B, CDK2AP1 and CHD3/4/5, which we refer to as the MGCC module, form the chromatin remodelling module of the nucleosome remodelling and deacetylase (NuRD) complex. To date, mechanisms by which the MGCC module acquires paralogue-specific function and specificity have not been addressed. Understanding the protein–protein interaction (PPI) network of the MGCC subunits is essential for defining underlying mechanisms of gene regulation. Therefore, using pulldown followed by mass spectrometry analysis (PD-MS), we report a proteome-wide interaction network of the MGCC module in a paralogue-specific manner. Our data also demonstrate that the disordered C-terminal region of CHD3/4/5 is a gateway to incorporate remodelling activity into both ChAHP (CHD4, ADNP, HP1γ) and NuRD complexes in a mutually exclusive manner. We define a short aggregation-prone region (APR) within the C-terminal segment of GATAD2B that is essential for the interaction of CHD4 and CDK2AP1 with the NuRD complex. Finally, we also report an association of CDK2AP1 with the nuclear receptor co-repressor (NCOR) complex. Overall, this study provides insight into the possible mechanisms through which the MGCC module can achieve specificity and diverse biological functions.},\\n bibtype = {article},\\n author = {Sharifi Tabar, M. and Giardina, C. and Feng, Y. and Francis, H. and Moghaddas Sani, H. and Low, J.K.K. and Mackay, J.P. and Bailey, C.G. and Rasko, J.E.J.},\\n doi = {10.1111/febs.16112},\\n journal = {FEBS Journal},\\n number = {1}\\n}\",\"author_short\":[\"Sharifi Tabar,  M.\",\"Giardina,  C.\",\"Feng,  Y.\",\"Francis,  H.\",\"Moghaddas Sani,  H.\",\"Low,  J.\",\"Mackay,  J.\",\"Bailey,  C.\",\"Rasko,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"sharifitabar-giardina-feng-francis-moghaddassani-low-mackay-bailey-etal-uniqueproteininteractionnetworksdefinethechromatinremodellingmoduleofthenurdcomplex-2022\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"BET-Family Bromodomains Can Recognize Diacetylated Sequences from Transcription Factors Using a Conserved Mechanism\",\"type\":\"article\",\"year\":\"2021\",\"pages\":\"648-662\",\"volume\":\"60\",\"id\":\"50cd9f3e-3d6d-35ea-b47c-1090ed52dff5\",\"created\":\"2023-01-10T01:43:48.713Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:48.713Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Almost all eukaryotic proteins receive diverse post-translational modifications (PTMs) that modulate protein activity. Many histone PTMs are well characterized, heavily influence gene regulation, and are often predictors of distinct transcriptional programs. Although our understanding of the histone PTM network has matured, much is yet to be understood about the roles of transcription factor (TF) PTMs, which might well represent a similarly complex and dynamic network of functional regulation. Members of the bromodomain and extra-terminal domain (BET) family of proteins recognize acetyllysine residues and relay the signals encoded by these modifications. Here, we have investigated the acetylation dependence of several functionally relevant BET-TF interactions in vitro using surface plasmon resonance, nuclear magnetic resonance, and X-ray crystallography. We show that motifs known to be acetylated in TFs E2F1 and MyoD1 can interact with all bromodomains of BRD2, BRD3, and BRD4. The interactions are dependent on diacetylation of the motifs and show a preference for the first BET bromodomain. Structural mapping of the interactions confirms a conserved mode of binding for the two TFs to the acetyllysine binding pocket of the BET bromodomains, mimicking that of other already established functionally important histone- and TF-BET interactions. We also examined a motif from the TF RelA that is known to be acetylated but were unable to observe any interaction, regardless of the acetylation state of the sequence. Our findings overall advance our understanding of BET-TF interactions and suggest a physical link between the important diacetylated motifs found in E2F1 and MyoD1 and the BET-family proteins.\",\"bibtype\":\"article\",\"author\":\"Patel, K. and Solomon, P.D. and Walshe, J.L. and Ford, D.J. and Wilkinson-White, L. and Payne, R.J. and Low, J.K.K. and Mackay, J.P.\",\"doi\":\"10.1021/acs.biochem.0c00816\",\"journal\":\"Biochemistry\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {BET-Family Bromodomains Can Recognize Diacetylated Sequences from Transcription Factors Using a Conserved Mechanism},\\n type = {article},\\n year = {2021},\\n pages = {648-662},\\n volume = {60},\\n id = {50cd9f3e-3d6d-35ea-b47c-1090ed52dff5},\\n created = {2023-01-10T01:43:48.713Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:48.713Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Almost all eukaryotic proteins receive diverse post-translational modifications (PTMs) that modulate protein activity. Many histone PTMs are well characterized, heavily influence gene regulation, and are often predictors of distinct transcriptional programs. Although our understanding of the histone PTM network has matured, much is yet to be understood about the roles of transcription factor (TF) PTMs, which might well represent a similarly complex and dynamic network of functional regulation. Members of the bromodomain and extra-terminal domain (BET) family of proteins recognize acetyllysine residues and relay the signals encoded by these modifications. Here, we have investigated the acetylation dependence of several functionally relevant BET-TF interactions in vitro using surface plasmon resonance, nuclear magnetic resonance, and X-ray crystallography. We show that motifs known to be acetylated in TFs E2F1 and MyoD1 can interact with all bromodomains of BRD2, BRD3, and BRD4. The interactions are dependent on diacetylation of the motifs and show a preference for the first BET bromodomain. Structural mapping of the interactions confirms a conserved mode of binding for the two TFs to the acetyllysine binding pocket of the BET bromodomains, mimicking that of other already established functionally important histone- and TF-BET interactions. We also examined a motif from the TF RelA that is known to be acetylated but were unable to observe any interaction, regardless of the acetylation state of the sequence. Our findings overall advance our understanding of BET-TF interactions and suggest a physical link between the important diacetylated motifs found in E2F1 and MyoD1 and the BET-family proteins.},\\n bibtype = {article},\\n author = {Patel, K. and Solomon, P.D. and Walshe, J.L. and Ford, D.J. and Wilkinson-White, L. and Payne, R.J. and Low, J.K.K. and Mackay, J.P.},\\n doi = {10.1021/acs.biochem.0c00816},\\n journal = {Biochemistry},\\n number = {9}\\n}\",\"author_short\":[\"Patel,  K.\",\"Solomon,  P.\",\"Walshe,  J.\",\"Ford,  D.\",\"Wilkinson-White,  L.\",\"Payne,  R.\",\"Low,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"patel-solomon-walshe-ford-wilkinsonwhite-payne-low-mackay-betfamilybromodomainscanrecognizediacetylatedsequencesfromtranscriptionfactorsusingaconservedmechanism-2021\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The characterization of protein interactions-what, how and how much?\",\"type\":\"article\",\"year\":\"2021\",\"pages\":\"12292-12307\",\"volume\":\"50\",\"id\":\"2d324f7d-025f-3a4a-af9f-b3ca4892be6b\",\"created\":\"2023-01-10T01:43:49.626Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:49.626Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Protein interactions underlie most molecular events in biology. Many methods have been developed to identify protein partners, to measure the affinity with which these biomolecules interact and to characterise the structures of the complexes. Each approach has its own advantages and limitations, and it can be difficult for the newcomer to determine which methodology would best suit their system. This review provides an overview of many of the techniques most widely used to identify protein partners, assess stoichiometry and binding affinity, and determine low-resolution models for complexes. Key methods covered include: yeast two-hybrid analysis, affinity purification mass spectrometry and proximity labelling to identify partners; size-exclusion chromatography, scattering methods, native mass spectrometry and analytical ultracentrifugation to estimate stoichiometry; isothermal titration calorimetry, biosensors and fluorometric methods (including microscale thermophoresis, anisotropy/polarisation, resonance energy transfer, AlphaScreen, and differential scanning fluorimetry) to measure binding affinity; and crosslinking and hydrogen-deuterium exchange mass spectrometry to probe the structure of complexes. This journal is\",\"bibtype\":\"article\",\"author\":\"Walport, L.J. and Low, J.K.K. and Matthews, J.M. and MacKay, J.P.\",\"doi\":\"10.1039/d1cs00548k\",\"journal\":\"Chemical Society Reviews\",\"number\":\"22\",\"bibtex\":\"@article{\\n title = {The characterization of protein interactions-what, how and how much?},\\n type = {article},\\n year = {2021},\\n pages = {12292-12307},\\n volume = {50},\\n id = {2d324f7d-025f-3a4a-af9f-b3ca4892be6b},\\n created = {2023-01-10T01:43:49.626Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:49.626Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Protein interactions underlie most molecular events in biology. Many methods have been developed to identify protein partners, to measure the affinity with which these biomolecules interact and to characterise the structures of the complexes. Each approach has its own advantages and limitations, and it can be difficult for the newcomer to determine which methodology would best suit their system. This review provides an overview of many of the techniques most widely used to identify protein partners, assess stoichiometry and binding affinity, and determine low-resolution models for complexes. Key methods covered include: yeast two-hybrid analysis, affinity purification mass spectrometry and proximity labelling to identify partners; size-exclusion chromatography, scattering methods, native mass spectrometry and analytical ultracentrifugation to estimate stoichiometry; isothermal titration calorimetry, biosensors and fluorometric methods (including microscale thermophoresis, anisotropy/polarisation, resonance energy transfer, AlphaScreen, and differential scanning fluorimetry) to measure binding affinity; and crosslinking and hydrogen-deuterium exchange mass spectrometry to probe the structure of complexes. This journal is},\\n bibtype = {article},\\n author = {Walport, L.J. and Low, J.K.K. and Matthews, J.M. and MacKay, J.P.},\\n doi = {10.1039/d1cs00548k},\\n journal = {Chemical Society Reviews},\\n number = {22}\\n}\",\"author_short\":[\"Walport,  L.\",\"Low,  J.\",\"Matthews,  J.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"walport-low-matthews-mackay-thecharacterizationofproteininteractionswhathowandhowmuch-2021\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":4,\"html\":\"\"},{\"title\":\"The bromodomains of BET family proteins can recognize diacetylated histone H2A.Z\",\"type\":\"article\",\"year\":\"2021\",\"pages\":\"464-476\",\"volume\":\"30\",\"id\":\"b502f967-797b-3fb0-8bb3-8923a5fa0bbd\",\"created\":\"2023-01-10T01:43:50.588Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:50.588Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Chemical modifications of histone tails influence genome accessibility and the transcriptional state of eukaryotic cells. Lysine acetylation is one of the most common modifications and acetyllysine-binding bromodomains (BDs) provide a means for acetyllysine marks to be translated into meaningful cellular responses. Here, we have investigated the mechanism underlying the reported association between the Bromodomain and Extra Terminal (BET) family of BD proteins and the essential histone variant H2A.Z. We use NMR spectroscopy to demonstrate a physical interaction between the N-terminal tail of H2A.Z and the BDs of BRD2, BRD3, and BRD4, and show that the interaction is dependent on lysine acetylation in H2A.Z. The BDs preferentially engage a diacetylated H2A.Z-K4acK7ac motif that is reminiscent of sequences found in other biologically important BET BD target proteins, including histones and transcription factors. A H2A.Z-K7acK11ac motif can also bind BET BDs—with a preference for the second BD of each protein. Chemical shift perturbation mapping of the interactions, together with an X-ray crystal structure of BRD2-BD1 bound to H2A.Z-K4acK7ac, shows that H2A.Z binds the canonical AcK binding pocket of the BDs. This mechanism mirrors the conserved binding mode that is unique to the BET BDs, in which two acetylation marks are read simultaneously by a single BD. Our findings provide structural corroboration of biochemical and cell biological data that link H2A.Z and BET-family proteins, suggesting that the function of H2A.Z is enacted through interactions with these chromatin readers.\",\"bibtype\":\"article\",\"author\":\"Patel, K. and Solomon, P.D. and Walshe, J.L. and Low, J.K.K. and Mackay, J.P.\",\"doi\":\"10.1002/pro.4006\",\"journal\":\"Protein Science\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {The bromodomains of BET family proteins can recognize diacetylated histone H2A.Z},\\n type = {article},\\n year = {2021},\\n pages = {464-476},\\n volume = {30},\\n id = {b502f967-797b-3fb0-8bb3-8923a5fa0bbd},\\n created = {2023-01-10T01:43:50.588Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:50.588Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Chemical modifications of histone tails influence genome accessibility and the transcriptional state of eukaryotic cells. Lysine acetylation is one of the most common modifications and acetyllysine-binding bromodomains (BDs) provide a means for acetyllysine marks to be translated into meaningful cellular responses. Here, we have investigated the mechanism underlying the reported association between the Bromodomain and Extra Terminal (BET) family of BD proteins and the essential histone variant H2A.Z. We use NMR spectroscopy to demonstrate a physical interaction between the N-terminal tail of H2A.Z and the BDs of BRD2, BRD3, and BRD4, and show that the interaction is dependent on lysine acetylation in H2A.Z. The BDs preferentially engage a diacetylated H2A.Z-K4acK7ac motif that is reminiscent of sequences found in other biologically important BET BD target proteins, including histones and transcription factors. A H2A.Z-K7acK11ac motif can also bind BET BDs—with a preference for the second BD of each protein. Chemical shift perturbation mapping of the interactions, together with an X-ray crystal structure of BRD2-BD1 bound to H2A.Z-K4acK7ac, shows that H2A.Z binds the canonical AcK binding pocket of the BDs. This mechanism mirrors the conserved binding mode that is unique to the BET BDs, in which two acetylation marks are read simultaneously by a single BD. Our findings provide structural corroboration of biochemical and cell biological data that link H2A.Z and BET-family proteins, suggesting that the function of H2A.Z is enacted through interactions with these chromatin readers.},\\n bibtype = {article},\\n author = {Patel, K. and Solomon, P.D. and Walshe, J.L. and Low, J.K.K. and Mackay, J.P.},\\n doi = {10.1002/pro.4006},\\n journal = {Protein Science},\\n number = {2}\\n}\",\"author_short\":[\"Patel,  K.\",\"Solomon,  P.\",\"Walshe,  J.\",\"Low,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"patel-solomon-walshe-low-mackay-thebromodomainsofbetfamilyproteinscanrecognizediacetylatedhistoneh2az-2021\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Late-stage modification of peptides and proteins at cysteine with diaryliodonium salts\",\"type\":\"article\",\"year\":\"2021\",\"pages\":\"14159-14166\",\"volume\":\"12\",\"id\":\"fe1847bf-a481-3fe2-9167-b70bf15b8ea0\",\"created\":\"2023-01-10T01:43:51.522Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:51.522Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The modification of peptides and proteins has emerged as a powerful means to efficiently prepare high value bioconjugates for a range of applications in chemical biology and for the development of next-generation therapeutics. Herein, we report a novel method for the chemoselective late-stage modification of peptides and proteins at cysteine in aqueous buffer with suitably functionalised diaryliodonium salts, furnishing stable thioether-linked synthetic conjugates. The power of this new platform is showcased through the late-stage modification of the affibody zEGFR and the histone protein H2A.\",\"bibtype\":\"article\",\"author\":\"Byrne, S.A. and Bedding, M.J. and Corcilius, L. and Ford, D.J. and Zhong, Y. and Franck, C. and Larance, M. and Mackay, J.P. and Payne, R.J.\",\"doi\":\"10.1039/d1sc03127a\",\"journal\":\"Chemical Science\",\"number\":\"42\",\"bibtex\":\"@article{\\n title = {Late-stage modification of peptides and proteins at cysteine with diaryliodonium salts},\\n type = {article},\\n year = {2021},\\n pages = {14159-14166},\\n volume = {12},\\n id = {fe1847bf-a481-3fe2-9167-b70bf15b8ea0},\\n created = {2023-01-10T01:43:51.522Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:51.522Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The modification of peptides and proteins has emerged as a powerful means to efficiently prepare high value bioconjugates for a range of applications in chemical biology and for the development of next-generation therapeutics. Herein, we report a novel method for the chemoselective late-stage modification of peptides and proteins at cysteine in aqueous buffer with suitably functionalised diaryliodonium salts, furnishing stable thioether-linked synthetic conjugates. The power of this new platform is showcased through the late-stage modification of the affibody zEGFR and the histone protein H2A.},\\n bibtype = {article},\\n author = {Byrne, S.A. and Bedding, M.J. and Corcilius, L. and Ford, D.J. and Zhong, Y. and Franck, C. and Larance, M. and Mackay, J.P. and Payne, R.J.},\\n doi = {10.1039/d1sc03127a},\\n journal = {Chemical Science},\\n number = {42}\\n}\",\"author_short\":[\"Byrne,  S.\",\"Bedding,  M.\",\"Corcilius,  L.\",\"Ford,  D.\",\"Zhong,  Y.\",\"Franck,  C.\",\"Larance,  M.\",\"Mackay,  J.\",\"Payne,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"byrne-bedding-corcilius-ford-zhong-franck-larance-mackay-etal-latestagemodificationofpeptidesandproteinsatcysteinewithdiaryliodoniumsalts-2021\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A single dose, BCG-adjuvanted COVID-19 vaccine provides sterilising immunity against SARS-CoV-2 infection\",\"type\":\"article\",\"year\":\"2021\",\"volume\":\"6\",\"id\":\"5f7017bd-f4d1-30e2-95a1-458fa2083a65\",\"created\":\"2023-01-10T01:43:52.472Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:52.472Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Global control of COVID-19 requires broadly accessible vaccines that are effective against SARS-CoV-2 variants. In this report, we exploit the immunostimulatory properties of bacille Calmette-Guérin (BCG), the existing tuberculosis vaccine, to deliver a vaccination regimen with potent SARS-CoV-2-specific protective immunity. Combination of BCG with a stabilised, trimeric form of SARS-CoV-2 spike antigen promoted rapid development of virus-specific IgG antibodies in the blood of vaccinated mice, that was further augmented by the addition of alum. This vaccine formulation, BCG:CoVac, induced high-titre SARS-CoV-2 neutralising antibodies (NAbs) and Th1-biased cytokine release by vaccine-specific T cells, which correlated with the early emergence of T follicular helper cells in local lymph nodes and heightened levels of antigen-specific plasma B cells after vaccination. Vaccination of K18-hACE2 mice with a single dose of BCG:CoVac almost completely abrogated disease after SARS-CoV-2 challenge, with minimal inflammation and no detectable virus in the lungs of infected animals. Boosting BCG:CoVac-primed mice with a heterologous vaccine further increased SARS-CoV-2-specific antibody responses, which effectively neutralised B.1.1.7 and B.1.351 SARS-CoV-2 variants of concern. These findings demonstrate the potential for BCG-based vaccination to protect against major SARS-CoV-2 variants circulating globally.\",\"bibtype\":\"article\",\"author\":\"Counoupas, C. and Johansen, M.D. and Stella, A.O. and Nguyen, D.H. and Ferguson, A.L. and Aggarwal, A. and Bhattacharyya, N.D. and Grey, A. and Hutchings, O. and Patel, K. and Hansbro, P.M. and Triccas, J.A.\",\"doi\":\"10.1038/s41541-021-00406-4\",\"journal\":\"npj Vaccines\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {A single dose, BCG-adjuvanted COVID-19 vaccine provides sterilising immunity against SARS-CoV-2 infection},\\n type = {article},\\n year = {2021},\\n volume = {6},\\n id = {5f7017bd-f4d1-30e2-95a1-458fa2083a65},\\n created = {2023-01-10T01:43:52.472Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:52.472Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Global control of COVID-19 requires broadly accessible vaccines that are effective against SARS-CoV-2 variants. In this report, we exploit the immunostimulatory properties of bacille Calmette-Guérin (BCG), the existing tuberculosis vaccine, to deliver a vaccination regimen with potent SARS-CoV-2-specific protective immunity. Combination of BCG with a stabilised, trimeric form of SARS-CoV-2 spike antigen promoted rapid development of virus-specific IgG antibodies in the blood of vaccinated mice, that was further augmented by the addition of alum. This vaccine formulation, BCG:CoVac, induced high-titre SARS-CoV-2 neutralising antibodies (NAbs) and Th1-biased cytokine release by vaccine-specific T cells, which correlated with the early emergence of T follicular helper cells in local lymph nodes and heightened levels of antigen-specific plasma B cells after vaccination. Vaccination of K18-hACE2 mice with a single dose of BCG:CoVac almost completely abrogated disease after SARS-CoV-2 challenge, with minimal inflammation and no detectable virus in the lungs of infected animals. Boosting BCG:CoVac-primed mice with a heterologous vaccine further increased SARS-CoV-2-specific antibody responses, which effectively neutralised B.1.1.7 and B.1.351 SARS-CoV-2 variants of concern. These findings demonstrate the potential for BCG-based vaccination to protect against major SARS-CoV-2 variants circulating globally.},\\n bibtype = {article},\\n author = {Counoupas, C. and Johansen, M.D. and Stella, A.O. and Nguyen, D.H. and Ferguson, A.L. and Aggarwal, A. and Bhattacharyya, N.D. and Grey, A. and Hutchings, O. and Patel, K. and Hansbro, P.M. and Triccas, J.A.},\\n doi = {10.1038/s41541-021-00406-4},\\n journal = {npj Vaccines},\\n number = {1}\\n}\",\"author_short\":[\"Counoupas,  C.\",\"Johansen,  M.\",\"Stella,  A.\",\"Nguyen,  D.\",\"Ferguson,  A.\",\"Aggarwal,  A.\",\"Bhattacharyya,  N.\",\"Grey,  A.\",\"Hutchings,  O.\",\"Patel,  K.\",\"Hansbro,  P.\",\"Triccas,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"counoupas-johansen-stella-nguyen-ferguson-aggarwal-bhattacharyya-grey-etal-asingledosebcgadjuvantedcovid19vaccineprovidessterilisingimmunityagainstsarscov2infection-2021\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Purification of an insect juvenile hormone receptor complex enables insights into its post-translational phosphorylation\",\"type\":\"article\",\"year\":\"2021\",\"volume\":\"297\",\"id\":\"f039ef2e-e6b3-3564-a85c-0dc612c27e87\",\"created\":\"2023-01-10T01:43:53.398Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:53.398Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Juvenile hormone (JH) plays vital roles in insect reproduction, development, and in many aspects of physiology. JH primarily acts at the gene-regulatory level through interaction with an intracellular receptor (JH receptor [JHR]), a ligand-activated complex of transcription factors consisting of the JH-binding protein methoprene-tolerant (MET) and its partner taiman (TAI). Initial studies indicated significance of post-transcriptional phosphorylation, subunit assembly, and nucleocytoplasmic transport of JHR in JH signaling. However, our knowledge of JHR regulation at the protein level remains rudimentary, partly because of the difficulty of obtaining purified and functional JHR proteins. Here, we present a method for high-yield expression and purification of JHR complexes from two insect species, the beetle T. castaneum and the mosquito Aedes aegypti. Recombinant JHR subunits from each species were coexpressed in an insect cell line using a baculovirus system. MET–TAI complexes were purified through affinity chromatography and anion exchange columns to yield proteins capable of binding both the hormonal ligand (JH III) and DNA bearing cognate JH-response elements. We further examined the beetle JHR complex in greater detail. Biochemical analyses and MS confirmed that T. castaneum JHR was a 1:1 heterodimer consisting of MET and Taiman proteins, stabilized by the JHR agonist ligand methoprene. Phosphoproteomics uncovered multiple phosphorylation sites in the MET protein, some of which were induced by methoprene treatment. Finally, we report a functional bipartite nuclear localization signal, straddled by phosphorylated residues, within the disordered C-terminal region of MET. Our present characterization of the recombinant JHR is an initial step toward understanding JHR structure and function.\",\"bibtype\":\"article\",\"author\":\"Jindra, M. and McKinstry, W.J. and Nebl, T. and Bittova, L. and Ren, B. and Shaw, J. and Phan, T. and Lu, L. and Low, J.K.K. and Mackay, J.P. and Lovrecz, G.O. and Hill, R.J.\",\"doi\":\"10.1016/j.jbc.2021.101387\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Purification of an insect juvenile hormone receptor complex enables insights into its post-translational phosphorylation},\\n type = {article},\\n year = {2021},\\n volume = {297},\\n id = {f039ef2e-e6b3-3564-a85c-0dc612c27e87},\\n created = {2023-01-10T01:43:53.398Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:53.398Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Juvenile hormone (JH) plays vital roles in insect reproduction, development, and in many aspects of physiology. JH primarily acts at the gene-regulatory level through interaction with an intracellular receptor (JH receptor [JHR]), a ligand-activated complex of transcription factors consisting of the JH-binding protein methoprene-tolerant (MET) and its partner taiman (TAI). Initial studies indicated significance of post-transcriptional phosphorylation, subunit assembly, and nucleocytoplasmic transport of JHR in JH signaling. However, our knowledge of JHR regulation at the protein level remains rudimentary, partly because of the difficulty of obtaining purified and functional JHR proteins. Here, we present a method for high-yield expression and purification of JHR complexes from two insect species, the beetle T. castaneum and the mosquito Aedes aegypti. Recombinant JHR subunits from each species were coexpressed in an insect cell line using a baculovirus system. MET–TAI complexes were purified through affinity chromatography and anion exchange columns to yield proteins capable of binding both the hormonal ligand (JH III) and DNA bearing cognate JH-response elements. We further examined the beetle JHR complex in greater detail. Biochemical analyses and MS confirmed that T. castaneum JHR was a 1:1 heterodimer consisting of MET and Taiman proteins, stabilized by the JHR agonist ligand methoprene. Phosphoproteomics uncovered multiple phosphorylation sites in the MET protein, some of which were induced by methoprene treatment. Finally, we report a functional bipartite nuclear localization signal, straddled by phosphorylated residues, within the disordered C-terminal region of MET. Our present characterization of the recombinant JHR is an initial step toward understanding JHR structure and function.},\\n bibtype = {article},\\n author = {Jindra, M. and McKinstry, W.J. and Nebl, T. and Bittova, L. and Ren, B. and Shaw, J. and Phan, T. and Lu, L. and Low, J.K.K. and Mackay, J.P. and Lovrecz, G.O. and Hill, R.J.},\\n doi = {10.1016/j.jbc.2021.101387},\\n journal = {Journal of Biological Chemistry},\\n number = {6}\\n}\",\"author_short\":[\"Jindra,  M.\",\"McKinstry,  W.\",\"Nebl,  T.\",\"Bittova,  L.\",\"Ren,  B.\",\"Shaw,  J.\",\"Phan,  T.\",\"Lu,  L.\",\"Low,  J.\",\"Mackay,  J.\",\"Lovrecz,  G.\",\"Hill,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"jindra-mckinstry-nebl-bittova-ren-shaw-phan-lu-etal-purificationofaninsectjuvenilehormonereceptorcomplexenablesinsightsintoitsposttranslationalphosphorylation-2021\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Discovery of Cyclic Peptide Ligands to the SARS-CoV-2 Spike Protein Using mRNA Display\",\"type\":\"article\",\"year\":\"2021\",\"pages\":\"1001-1008\",\"volume\":\"7\",\"id\":\"278be5c6-73f1-3344-9bd1-51ffab0169e6\",\"created\":\"2023-01-10T01:43:54.314Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:54.314Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The COVID-19 pandemic, caused by SARS-CoV-2, has led to substantial morbidity, mortality, and disruption globally. Cellular entry of SARS-CoV-2 is mediated by the viral spike protein, and affinity ligands to this surface protein have the potential for applications as antivirals and diagnostic reagents. Here, we describe the affinity selection of cyclic peptide ligands to the SARS-CoV-2 spike protein receptor binding domain (RBD) from three distinct libraries (in excess of a trillion molecules each) by mRNA display. We identified six high affinity molecules with dissociation constants (KD) in the nanomolar range (15-550 nM) to the RBD. The highest affinity ligand could be used as an affinity reagent to detect the spike protein in solution by ELISA, and the cocrystal structure of this molecule bound to the RBD demonstrated that it binds to a cryptic binding site, displacing a β-strand near the C-terminus. Our findings provide key mechanistic insight into the binding of peptide ligands to the SARS-CoV-2 spike RBD, and the ligands discovered in this work may find future use as reagents for diagnostic applications.\",\"bibtype\":\"article\",\"author\":\"Norman, A. and Franck, C. and Christie, M. and Hawkins, P.M.E. and Patel, K. and Ashhurst, A.S. and Aggarwal, A. and Low, J.K.K. and Siddiquee, R. and Ashley, C.L. and Passioura, T. and Payne, R.J.\",\"doi\":\"10.1021/acscentsci.0c01708\",\"journal\":\"ACS Central Science\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Discovery of Cyclic Peptide Ligands to the SARS-CoV-2 Spike Protein Using mRNA Display},\\n type = {article},\\n year = {2021},\\n pages = {1001-1008},\\n volume = {7},\\n id = {278be5c6-73f1-3344-9bd1-51ffab0169e6},\\n created = {2023-01-10T01:43:54.314Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:54.314Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The COVID-19 pandemic, caused by SARS-CoV-2, has led to substantial morbidity, mortality, and disruption globally. Cellular entry of SARS-CoV-2 is mediated by the viral spike protein, and affinity ligands to this surface protein have the potential for applications as antivirals and diagnostic reagents. Here, we describe the affinity selection of cyclic peptide ligands to the SARS-CoV-2 spike protein receptor binding domain (RBD) from three distinct libraries (in excess of a trillion molecules each) by mRNA display. We identified six high affinity molecules with dissociation constants (KD) in the nanomolar range (15-550 nM) to the RBD. The highest affinity ligand could be used as an affinity reagent to detect the spike protein in solution by ELISA, and the cocrystal structure of this molecule bound to the RBD demonstrated that it binds to a cryptic binding site, displacing a β-strand near the C-terminus. Our findings provide key mechanistic insight into the binding of peptide ligands to the SARS-CoV-2 spike RBD, and the ligands discovered in this work may find future use as reagents for diagnostic applications.},\\n bibtype = {article},\\n author = {Norman, A. and Franck, C. and Christie, M. and Hawkins, P.M.E. and Patel, K. and Ashhurst, A.S. and Aggarwal, A. and Low, J.K.K. and Siddiquee, R. and Ashley, C.L. and Passioura, T. and Payne, R.J.},\\n doi = {10.1021/acscentsci.0c01708},\\n journal = {ACS Central Science},\\n number = {6}\\n}\",\"author_short\":[\"Norman,  A.\",\"Franck,  C.\",\"Christie,  M.\",\"Hawkins,  P.\",\"Patel,  K.\",\"Ashhurst,  A.\",\"Aggarwal,  A.\",\"Low,  J.\",\"Siddiquee,  R.\",\"Ashley,  C.\",\"Passioura,  T.\",\"Payne,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"norman-franck-christie-hawkins-patel-ashhurst-aggarwal-low-etal-discoveryofcyclicpeptideligandstothesarscov2spikeproteinusingmrnadisplay-2021\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":4,\"html\":\"\"},{\"title\":\"GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder\",\"type\":\"article\",\"year\":\"2020\",\"pages\":\"878-888\",\"volume\":\"22\",\"id\":\"85308366-aa04-3a25-93ec-e3edc0ad78cc\",\"created\":\"2023-01-10T01:43:55.280Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:55.280Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.\",\"bibtype\":\"article\",\"author\":\"Shieh, C. and Jones, N. and Vanle, B. and Au, M. and Huang, A.Y. and Silva, A.P.G. and Lee, H. and Douine, E.D. and Otero, M.G. and Choi, A. and Mackay, J.P. and Pierson, T.M.\",\"doi\":\"10.1038/s41436-019-0747-z\",\"journal\":\"Genetics in Medicine\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder},\\n type = {article},\\n year = {2020},\\n pages = {878-888},\\n volume = {22},\\n id = {85308366-aa04-3a25-93ec-e3edc0ad78cc},\\n created = {2023-01-10T01:43:55.280Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:55.280Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.},\\n bibtype = {article},\\n author = {Shieh, C. and Jones, N. and Vanle, B. and Au, M. and Huang, A.Y. and Silva, A.P.G. and Lee, H. and Douine, E.D. and Otero, M.G. and Choi, A. and Mackay, J.P. and Pierson, T.M.},\\n doi = {10.1038/s41436-019-0747-z},\\n journal = {Genetics in Medicine},\\n number = {5}\\n}\",\"author_short\":[\"Shieh,  C.\",\"Jones,  N.\",\"Vanle,  B.\",\"Au,  M.\",\"Huang,  A.\",\"Silva,  A.\",\"Lee,  H.\",\"Douine,  E.\",\"Otero,  M.\",\"Choi,  A.\",\"Mackay,  J.\",\"Pierson,  T.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Correction: Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system.(Applied Optics(2020)295(1898–1914)Doi: 10.1074/jbc.RA119.010679)\",\"type\":\"article\",\"year\":\"2020\",\"pages\":\"6251\",\"volume\":\"295\",\"id\":\"9fc40b26-4df6-3126-8c8e-92fc8825b167\",\"created\":\"2023-01-10T01:43:56.242Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:56.242Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Yichen Zhong's name was misspelled. The correct spelling is shown above.\",\"bibtype\":\"article\",\"author\":\"Werner, M.T. and Wang, H. and Hamagami, N. and Hsu, S.C. and Yano, J.A. and Stonestrom, A.J. and Behera, V. and Zhong, Y. and MacKay, J.P. and Blobel, G.A.\",\"doi\":\"10.1074/jbc.AAC120.013771\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"18\",\"bibtex\":\"@article{\\n title = {Correction: Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system.(Applied Optics(2020)295(1898–1914)Doi: 10.1074/jbc.RA119.010679)},\\n type = {article},\\n year = {2020},\\n pages = {6251},\\n volume = {295},\\n id = {9fc40b26-4df6-3126-8c8e-92fc8825b167},\\n created = {2023-01-10T01:43:56.242Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:56.242Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Yichen Zhong's name was misspelled. The correct spelling is shown above.},\\n bibtype = {article},\\n author = {Werner, M.T. and Wang, H. and Hamagami, N. and Hsu, S.C. and Yano, J.A. and Stonestrom, A.J. and Behera, V. and Zhong, Y. and MacKay, J.P. and Blobel, G.A.},\\n doi = {10.1074/jbc.AAC120.013771},\\n journal = {Journal of Biological Chemistry},\\n number = {18}\\n}\",\"author_short\":[\"Werner,  M.\",\"Wang,  H.\",\"Hamagami,  N.\",\"Hsu,  S.\",\"Yano,  J.\",\"Stonestrom,  A.\",\"Behera,  V.\",\"Zhong,  Y.\",\"MacKay,  J.\",\"Blobel,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"werner-wang-hamagami-hsu-yano-stonestrom-behera-zhong-etal-correctioncomparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystemappliedoptics202029518981914doi101074jbcra119010679-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Cyclic peptides can engage a single binding pocket through highly divergent modes\",\"type\":\"article\",\"year\":\"2020\",\"pages\":\"26728-26738\",\"volume\":\"117\",\"id\":\"9d68a322-e2a2-3b30-b21a-47d62f211339\",\"created\":\"2023-01-10T01:43:57.164Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:57.164Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.\",\"bibtype\":\"article\",\"author\":\"Patel, K. and Walport, L.J. and Walshe, J.L. and Solomon, P.D. and Low, J.K.K. and Tran, D.H. and Mouradian, K.S. and Silva, A.P.G. and Wilkinson-White, L. and Norman, A. and Suga, H. and Mackay, J.P.\",\"doi\":\"10.1073/pnas.2003086117\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"43\",\"bibtex\":\"@article{\\n title = {Cyclic peptides can engage a single binding pocket through highly divergent modes},\\n type = {article},\\n year = {2020},\\n pages = {26728-26738},\\n volume = {117},\\n id = {9d68a322-e2a2-3b30-b21a-47d62f211339},\\n created = {2023-01-10T01:43:57.164Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:57.164Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.},\\n bibtype = {article},\\n author = {Patel, K. and Walport, L.J. and Walshe, J.L. and Solomon, P.D. and Low, J.K.K. and Tran, D.H. and Mouradian, K.S. and Silva, A.P.G. and Wilkinson-White, L. and Norman, A. and Suga, H. and Mackay, J.P.},\\n doi = {10.1073/pnas.2003086117},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {43}\\n}\",\"author_short\":[\"Patel,  K.\",\"Walport,  L.\",\"Walshe,  J.\",\"Solomon,  P.\",\"Low,  J.\",\"Tran,  D.\",\"Mouradian,  K.\",\"Silva,  A.\",\"Wilkinson-White,  L.\",\"Norman,  A.\",\"Suga,  H.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Semisynthesis of an evasin from tick saliva reveals a critical role of tyrosine sulfation for chemokine binding and inhibition\",\"type\":\"article\",\"year\":\"2020\",\"pages\":\"12657-12664\",\"volume\":\"117\",\"id\":\"b8c84666-fc07-3ad8-aaf9-d251ef3b0ee8\",\"created\":\"2023-01-10T01:43:58.083Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:58.083Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Blood-feeding arthropods produce antiinflammatory salivary proteins called evasins that function through inhibition of chemokine-receptor signaling in the host. Herein, we show that the evasin ACA-01 from the Amblyomma cajennense tick can be posttranslationally sulfated at two tyrosine residues, albeit as a mixture of sulfated variants. Homogenously sulfated variants of the proteins were efficiently assembled via a semisynthetic native chemical ligation strategy. Sulfation significantly improved the binding affinity of ACA-01 for a range of proinflammatory chemokines and enhanced the ability of ACA-01 to inhibit chemokine signaling through cognate receptors. Comparisons of evasin sequences and structural data suggest that tyrosine sulfation serves as a receptor mimetic strategy for recognizing and suppressing the proinflammatory activity of a wide variety of mammalian chemokines. As such, the incorporation of this posttranslational modification (PTM) or mimics thereof into evasins may provide a strategy to optimize tick salivary proteins for antiinflammatory applications.\",\"bibtype\":\"article\",\"author\":\"Franck, C. and Foster, S.R. and Johansen-Leete, J. and Chowdhury, S. and Cielesh, M. and Bhusal, R.P. and Mackay, J.P. and Larance, M. and Stone, M.J. and Payne, R.J.\",\"doi\":\"10.1073/pnas.2000605117\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"23\",\"bibtex\":\"@article{\\n title = {Semisynthesis of an evasin from tick saliva reveals a critical role of tyrosine sulfation for chemokine binding and inhibition},\\n type = {article},\\n year = {2020},\\n pages = {12657-12664},\\n volume = {117},\\n id = {b8c84666-fc07-3ad8-aaf9-d251ef3b0ee8},\\n created = {2023-01-10T01:43:58.083Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:58.083Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Blood-feeding arthropods produce antiinflammatory salivary proteins called evasins that function through inhibition of chemokine-receptor signaling in the host. Herein, we show that the evasin ACA-01 from the Amblyomma cajennense tick can be posttranslationally sulfated at two tyrosine residues, albeit as a mixture of sulfated variants. Homogenously sulfated variants of the proteins were efficiently assembled via a semisynthetic native chemical ligation strategy. Sulfation significantly improved the binding affinity of ACA-01 for a range of proinflammatory chemokines and enhanced the ability of ACA-01 to inhibit chemokine signaling through cognate receptors. Comparisons of evasin sequences and structural data suggest that tyrosine sulfation serves as a receptor mimetic strategy for recognizing and suppressing the proinflammatory activity of a wide variety of mammalian chemokines. As such, the incorporation of this posttranslational modification (PTM) or mimics thereof into evasins may provide a strategy to optimize tick salivary proteins for antiinflammatory applications.},\\n bibtype = {article},\\n author = {Franck, C. and Foster, S.R. and Johansen-Leete, J. and Chowdhury, S. and Cielesh, M. and Bhusal, R.P. and Mackay, J.P. and Larance, M. and Stone, M.J. and Payne, R.J.},\\n doi = {10.1073/pnas.2000605117},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {23}\\n}\",\"author_short\":[\"Franck,  C.\",\"Foster,  S.\",\"Johansen-Leete,  J.\",\"Chowdhury,  S.\",\"Cielesh,  M.\",\"Bhusal,  R.\",\"Mackay,  J.\",\"Larance,  M.\",\"Stone,  M.\",\"Payne,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"franck-foster-johansenleete-chowdhury-cielesh-bhusal-mackay-larance-etal-semisynthesisofanevasinfromticksalivarevealsacriticalroleoftyrosinesulfationforchemokinebindingandinhibition-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":4,\"html\":\"\"},{\"title\":\"A Novel Purification Procedure for Active Recombinant Human DPP4 and the Inability of DPP4 to Bind SARS-CoV-2\",\"type\":\"article\",\"year\":\"2020\",\"volume\":\"25\",\"id\":\"016baf4e-9b3e-39c6-87c2-be2e11c5cbaa\",\"created\":\"2023-01-10T01:43:59.005Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:59.005Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Proteases catalyse irreversible posttranslational modifications that often alter a biological function of the substrate. The protease dipeptidyl peptidase 4 (DPP4) is a pharmacological target in type 2 diabetes therapy primarily because it inactivates glucagon-like protein-1. DPP4 also has roles in steatosis, insulin resistance, cancers and inflammatory and fibrotic diseases. In addition, DPP4 binds to the spike protein of the MERS virus, causing it to be the human cell surface receptor for that virus. DPP4 has been identified as a potential binding target of SARS-CoV-2 spike protein, so this question requires experimental investigation. Understanding protein structure and function requires reliable protocols for production and purification. We developed such strategies for baculovirus generated soluble recombinant human DPP4 (residues 29–766) produced in insect cells. Purification used differential ammonium sulphate precipitation, hydrophobic interaction chromatography, dye affinity chromatography in series with immobilised metal affinity chromatography, and ion-exchange chromatography. The binding affinities of DPP4 to the SARS-CoV-2 full-length spike protein and its receptor-binding domain (RBD) were measured using surface plasmon resonance and ELISA. This optimised DPP4 purification procedure yielded 1 to 1.8 mg of pure fully active soluble DPP4 protein per litre of insect cell culture with specific activity >30 U/mg, indicative of high purity. No specific binding between DPP4 and CoV-2 spike protein was detected by surface plasmon resonance or ELISA. In summary, a procedure for high purity high yield soluble human DPP4 was achieved and used to show that, unlike MERS, SARS-CoV-2 does not bind human DPP4.\",\"bibtype\":\"article\",\"author\":\"Xi, C.R. and Di Fazio, A. and Nadvi, N.A. and Patel, K. and Xiang, M.S.W. and Zhang, H.E. and Deshpande, C. and Low, J.K.K. and Wang, X.T. and Chen, Y. and Church, W.B. and Gorrell, M.D.\",\"doi\":\"10.3390/MOLECULES25225392\",\"journal\":\"Molecules\",\"number\":\"22\",\"bibtex\":\"@article{\\n title = {A Novel Purification Procedure for Active Recombinant Human DPP4 and the Inability of DPP4 to Bind SARS-CoV-2},\\n type = {article},\\n year = {2020},\\n volume = {25},\\n id = {016baf4e-9b3e-39c6-87c2-be2e11c5cbaa},\\n created = {2023-01-10T01:43:59.005Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:59.005Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Proteases catalyse irreversible posttranslational modifications that often alter a biological function of the substrate. The protease dipeptidyl peptidase 4 (DPP4) is a pharmacological target in type 2 diabetes therapy primarily because it inactivates glucagon-like protein-1. DPP4 also has roles in steatosis, insulin resistance, cancers and inflammatory and fibrotic diseases. In addition, DPP4 binds to the spike protein of the MERS virus, causing it to be the human cell surface receptor for that virus. DPP4 has been identified as a potential binding target of SARS-CoV-2 spike protein, so this question requires experimental investigation. Understanding protein structure and function requires reliable protocols for production and purification. We developed such strategies for baculovirus generated soluble recombinant human DPP4 (residues 29–766) produced in insect cells. Purification used differential ammonium sulphate precipitation, hydrophobic interaction chromatography, dye affinity chromatography in series with immobilised metal affinity chromatography, and ion-exchange chromatography. The binding affinities of DPP4 to the SARS-CoV-2 full-length spike protein and its receptor-binding domain (RBD) were measured using surface plasmon resonance and ELISA. This optimised DPP4 purification procedure yielded 1 to 1.8 mg of pure fully active soluble DPP4 protein per litre of insect cell culture with specific activity >30 U/mg, indicative of high purity. No specific binding between DPP4 and CoV-2 spike protein was detected by surface plasmon resonance or ELISA. In summary, a procedure for high purity high yield soluble human DPP4 was achieved and used to show that, unlike MERS, SARS-CoV-2 does not bind human DPP4.},\\n bibtype = {article},\\n author = {Xi, C.R. and Di Fazio, A. and Nadvi, N.A. and Patel, K. and Xiang, M.S.W. and Zhang, H.E. and Deshpande, C. and Low, J.K.K. and Wang, X.T. and Chen, Y. and Church, W.B. and Gorrell, M.D.},\\n doi = {10.3390/MOLECULES25225392},\\n journal = {Molecules},\\n number = {22}\\n}\",\"author_short\":[\"Xi,  C.\",\"Di Fazio,  A.\",\"Nadvi,  N.\",\"Patel,  K.\",\"Xiang,  M.\",\"Zhang,  H.\",\"Deshpande,  C.\",\"Low,  J.\",\"Wang,  X.\",\"Chen,  Y.\",\"Church,  W.\",\"Gorrell,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"xi-difazio-nadvi-patel-xiang-zhang-deshpande-low-etal-anovelpurificationprocedureforactiverecombinanthumandpp4andtheinabilityofdpp4tobindsarscov2-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"CHD4 slides nucleosomes by decoupling entry- and exit-side DNA translocation\",\"type\":\"article\",\"year\":\"2020\",\"volume\":\"11\",\"id\":\"5dc9783b-933a-3c63-9cd9-b4ae4e6fd62e\",\"created\":\"2023-01-10T01:43:59.935Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:43:59.935Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Chromatin remodellers hydrolyse ATP to move nucleosomal DNA against histone octamers. The mechanism, however, is only partially resolved, and it is unclear if it is conserved among the four remodeller families. Here we use single-molecule assays to examine the mechanism of action of CHD4, which is part of the least well understood family. We demonstrate that the binding energy for CHD4-nucleosome complex formation—even in the absence of nucleotide—triggers significant conformational changes in DNA at the entry side, effectively priming the system for remodelling. During remodelling, flanking DNA enters the nucleosome in a continuous, gradual manner but exits in concerted 4–6 base-pair steps. This decoupling of entry- and exit-side translocation suggests that ATP-driven movement of entry-side DNA builds up strain inside the nucleosome that is subsequently released at the exit side by DNA expulsion. Based on our work and previous studies, we propose a mechanism for nucleosome sliding.\",\"bibtype\":\"article\",\"author\":\"Zhong, Y. and Paudel, B.P. and Ryan, D.P. and Low, J.K.K. and Franck, C. and Patel, K. and Bedward, M.J. and Torrado, M. and Payne, R.J. and van Oijen, A.M. and van Oijen, A.M. and Mackay, J.P.\",\"doi\":\"10.1038/s41467-020-15183-2\",\"journal\":\"Nature Communications\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {CHD4 slides nucleosomes by decoupling entry- and exit-side DNA translocation},\\n type = {article},\\n year = {2020},\\n volume = {11},\\n id = {5dc9783b-933a-3c63-9cd9-b4ae4e6fd62e},\\n created = {2023-01-10T01:43:59.935Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:43:59.935Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Chromatin remodellers hydrolyse ATP to move nucleosomal DNA against histone octamers. The mechanism, however, is only partially resolved, and it is unclear if it is conserved among the four remodeller families. Here we use single-molecule assays to examine the mechanism of action of CHD4, which is part of the least well understood family. We demonstrate that the binding energy for CHD4-nucleosome complex formation—even in the absence of nucleotide—triggers significant conformational changes in DNA at the entry side, effectively priming the system for remodelling. During remodelling, flanking DNA enters the nucleosome in a continuous, gradual manner but exits in concerted 4–6 base-pair steps. This decoupling of entry- and exit-side translocation suggests that ATP-driven movement of entry-side DNA builds up strain inside the nucleosome that is subsequently released at the exit side by DNA expulsion. Based on our work and previous studies, we propose a mechanism for nucleosome sliding.},\\n bibtype = {article},\\n author = {Zhong, Y. and Paudel, B.P. and Ryan, D.P. and Low, J.K.K. and Franck, C. and Patel, K. and Bedward, M.J. and Torrado, M. and Payne, R.J. and van Oijen, A.M. and van Oijen, A.M. and Mackay, J.P.},\\n doi = {10.1038/s41467-020-15183-2},\\n journal = {Nature Communications},\\n number = {1}\\n}\",\"author_short\":[\"Zhong,  Y.\",\"Paudel,  B.\",\"Ryan,  D.\",\"Low,  J.\",\"Franck,  C.\",\"Patel,  K.\",\"Bedward,  M.\",\"Torrado,  M.\",\"Payne,  R.\",\"van Oijen,  A.\",\"van Oijen,  A.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"zhong-paudel-ryan-low-franck-patel-bedward-torrado-etal-chd4slidesnucleosomesbydecouplingentryandexitsidednatranslocation-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"A heme-binding protein produced by Haemophilus haemolyticus inhibits non-typeable Haemophilus influenzae\",\"type\":\"article\",\"year\":\"2020\",\"pages\":\"381-398\",\"volume\":\"113\",\"id\":\"9a9059a4-c040-3b6b-8ed2-e2bf25cedaa5\",\"created\":\"2023-01-10T01:44:00.856Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:00.856Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Commensal bacteria serve as an important line of defense against colonisation by opportunisitic pathogens, but the underlying molecular mechanisms remain poorly explored. Here, we show that strains of a commensal bacterium, Haemophilus haemolyticus, make hemophilin, a heme-binding protein that inhibits growth of the opportunistic pathogen, non-typeable Haemophilus influenzae (NTHi) in culture. We purified the NTHi-inhibitory protein from H. haemolyticus and identified the hemophilin gene using proteomics and a gene knockout. An x-ray crystal structure of recombinant hemophilin shows that the protein does not belong to any of the known heme-binding protein folds, suggesting that it evolved independently. Biochemical characterisation shows that heme can be captured in the ferrous or ferric state, and with a variety of small heme-ligands bound, suggesting that hemophilin could function under a range of physiological conditions. Hemophilin knockout bacteria show a limited capacity to utilise free heme for growth. Our data suggest that hemophilin is a hemophore and that inhibition of NTHi occurs by heme starvation, raising the possibility that competition from hemophilin-producing H. haemolyticus could antagonise NTHi colonisation in the respiratory tract.\",\"bibtype\":\"article\",\"author\":\"Latham, R.D. and Torrado, M. and Atto, B. and Walshe, J.L. and Wilson, R. and Guss, J.M. and Mackay, J.P. and Tristram, S. and Gell, D.A.\",\"doi\":\"10.1111/mmi.14426\",\"journal\":\"Molecular Microbiology\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {A heme-binding protein produced by Haemophilus haemolyticus inhibits non-typeable Haemophilus influenzae},\\n type = {article},\\n year = {2020},\\n pages = {381-398},\\n volume = {113},\\n id = {9a9059a4-c040-3b6b-8ed2-e2bf25cedaa5},\\n created = {2023-01-10T01:44:00.856Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:00.856Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Commensal bacteria serve as an important line of defense against colonisation by opportunisitic pathogens, but the underlying molecular mechanisms remain poorly explored. Here, we show that strains of a commensal bacterium, Haemophilus haemolyticus, make hemophilin, a heme-binding protein that inhibits growth of the opportunistic pathogen, non-typeable Haemophilus influenzae (NTHi) in culture. We purified the NTHi-inhibitory protein from H. haemolyticus and identified the hemophilin gene using proteomics and a gene knockout. An x-ray crystal structure of recombinant hemophilin shows that the protein does not belong to any of the known heme-binding protein folds, suggesting that it evolved independently. Biochemical characterisation shows that heme can be captured in the ferrous or ferric state, and with a variety of small heme-ligands bound, suggesting that hemophilin could function under a range of physiological conditions. Hemophilin knockout bacteria show a limited capacity to utilise free heme for growth. Our data suggest that hemophilin is a hemophore and that inhibition of NTHi occurs by heme starvation, raising the possibility that competition from hemophilin-producing H. haemolyticus could antagonise NTHi colonisation in the respiratory tract.},\\n bibtype = {article},\\n author = {Latham, R.D. and Torrado, M. and Atto, B. and Walshe, J.L. and Wilson, R. and Guss, J.M. and Mackay, J.P. and Tristram, S. and Gell, D.A.},\\n doi = {10.1111/mmi.14426},\\n journal = {Molecular Microbiology},\\n number = {2}\\n}\",\"author_short\":[\"Latham,  R.\",\"Torrado,  M.\",\"Atto,  B.\",\"Walshe,  J.\",\"Wilson,  R.\",\"Guss,  J.\",\"Mackay,  J.\",\"Tristram,  S.\",\"Gell,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"latham-torrado-atto-walshe-wilson-guss-mackay-tristram-etal-ahemebindingproteinproducedbyhaemophilushaemolyticusinhibitsnontypeablehaemophilusinfluenzae-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":4,\"html\":\"\"},{\"title\":\"Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system\",\"type\":\"article\",\"year\":\"2020\",\"pages\":\"1898-1914\",\"volume\":\"295\",\"id\":\"53a92f38-e8f3-3c7c-9fcf-6b8b0bb9521a\",\"created\":\"2023-01-10T01:44:01.787Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:01.787Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The widely expressed bromodomain and extraterminal motif (BET) proteins bromodomain-containing protein 2 (BRD2), BRD3, and BRD4 are multifunctional transcriptional regulators that bind acetylated chromatin via their conserved tandem bromodomains. Small molecules that target BET bromodomains are being tested for various diseases but typically do not discern between BET family members. Genomic distributions and protein partners of BET proteins have been described, but the basis for differences in BET protein function within a given lineage remains unclear. By establishing a gene knockout-rescue system in a Brd2-null erythroblast cell line, here we compared a series of mutant and chimeric BET proteins for their ability to modulate cell growth, differentiation, and gene expression. We found that the BET N-terminal halves bearing the bromodomains convey marked differences in protein stability but do not account for specificity in BET protein function. Instead, when BET proteins were expressed at comparable levels, their specificity was largely determined by the C-terminal half. Remarkably, a chimeric BET protein comprising the N-terminal half of the structurally similar short BRD4 isoform (BRD4S) and the C-terminal half of BRD2 functioned similarly to intact BRD2. We traced part of the BRD2-specific activity to a previously uncharacterized short segment predicted to harbor a coiled-coil (CC) domain. Deleting the CC segment impaired BRD2's ability to restore growth and differentiation, and the CC region functioned in conjunction with the adjacent ET domain to impart BRD2-like activity onto BRD4S. In summary, our results identify distinct BET protein domains that regulate protein turnover and biological activities.\",\"bibtype\":\"article\",\"author\":\"Werner, M.T. and Wang, H. and Hamagami, N. and Hsu, S.C. and Yano, J.A. and Stonestrom, A.J. and Behera, V. and Zong, Y. and Mackay, J.P. and Blobel, G.A.\",\"doi\":\"10.1074/jbc.RA119.010679\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system},\\n type = {article},\\n year = {2020},\\n pages = {1898-1914},\\n volume = {295},\\n id = {53a92f38-e8f3-3c7c-9fcf-6b8b0bb9521a},\\n created = {2023-01-10T01:44:01.787Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:01.787Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The widely expressed bromodomain and extraterminal motif (BET) proteins bromodomain-containing protein 2 (BRD2), BRD3, and BRD4 are multifunctional transcriptional regulators that bind acetylated chromatin via their conserved tandem bromodomains. Small molecules that target BET bromodomains are being tested for various diseases but typically do not discern between BET family members. Genomic distributions and protein partners of BET proteins have been described, but the basis for differences in BET protein function within a given lineage remains unclear. By establishing a gene knockout-rescue system in a Brd2-null erythroblast cell line, here we compared a series of mutant and chimeric BET proteins for their ability to modulate cell growth, differentiation, and gene expression. We found that the BET N-terminal halves bearing the bromodomains convey marked differences in protein stability but do not account for specificity in BET protein function. Instead, when BET proteins were expressed at comparable levels, their specificity was largely determined by the C-terminal half. Remarkably, a chimeric BET protein comprising the N-terminal half of the structurally similar short BRD4 isoform (BRD4S) and the C-terminal half of BRD2 functioned similarly to intact BRD2. We traced part of the BRD2-specific activity to a previously uncharacterized short segment predicted to harbor a coiled-coil (CC) domain. Deleting the CC segment impaired BRD2's ability to restore growth and differentiation, and the CC region functioned in conjunction with the adjacent ET domain to impart BRD2-like activity onto BRD4S. In summary, our results identify distinct BET protein domains that regulate protein turnover and biological activities.},\\n bibtype = {article},\\n author = {Werner, M.T. and Wang, H. and Hamagami, N. and Hsu, S.C. and Yano, J.A. and Stonestrom, A.J. and Behera, V. and Zong, Y. and Mackay, J.P. and Blobel, G.A.},\\n doi = {10.1074/jbc.RA119.010679},\\n journal = {Journal of Biological Chemistry},\\n number = {7}\\n}\",\"author_short\":[\"Werner,  M.\",\"Wang,  H.\",\"Hamagami,  N.\",\"Hsu,  S.\",\"Yano,  J.\",\"Stonestrom,  A.\",\"Behera,  V.\",\"Zong,  Y.\",\"Mackay,  J.\",\"Blobel,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"werner-wang-hamagami-hsu-yano-stonestrom-behera-zong-etal-comparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystem-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Thermostable small-molecule inhibitor of angiogenesis and vascular permeability that suppresses a pERK-FosB/ΔFosB-VCAM-1 axis\",\"type\":\"article\",\"year\":\"2020\",\"volume\":\"6\",\"id\":\"f819966b-5abf-3066-bb8c-9914a970f868\",\"created\":\"2023-01-10T01:44:02.733Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:02.733Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Vascular permeability and angiogenesis underpin neovascular age-related macular degeneration and diabetic retinopathy. While anti-VEGF therapies are widely used clinically, many patients do not respond optimally, or at all, and small-molecule therapies are lacking. Here, we identified a dibenzoxazepinone BT2 that inhibits endothelial cell proliferation, migration, wound repair in vitro, network formation, and angiogenesis in mice bearing Matrigel plugs. BT2 interacts with MEK1 and inhibits ERK phosphorylation and the expression of FosB/ΔFosB, VCAM-1, and many genes involved in proliferation, migration, angiogenesis, and inflammation. BT2 reduced retinal vascular leakage following rat choroidal laser trauma and rabbit intravitreal VEGF-A165 administration. BT2 suppressed retinal CD31, pERK, VCAM-1, and VEGF-A165 expression. BT2 reduced retinal leakage in rats at least as effectively as aflibercept, a first-line therapy for nAMD/DR. BT2 withstands boiling or autoclaving and several months' storage at 22°C. BT2 is a new small-molecule inhibitor of vascular permeability and angiogenesis.\",\"bibtype\":\"article\",\"author\":\"Li, Y. and Alhendi, A.M.N. and Yeh, M.-C. and Elahy, M. and Santiago, F.S. and Deshpande, N.P. and Wu, B. and Chan, E. and Inam, S. and Prado-Lourenco, L. and Marcuccio, S.M. and Khachigian, L.M.\",\"doi\":\"10.1126/sciadv.aaz7815\",\"journal\":\"Science Advances\",\"number\":\"31\",\"bibtex\":\"@article{\\n title = {Thermostable small-molecule inhibitor of angiogenesis and vascular permeability that suppresses a pERK-FosB/ΔFosB-VCAM-1 axis},\\n type = {article},\\n year = {2020},\\n volume = {6},\\n id = {f819966b-5abf-3066-bb8c-9914a970f868},\\n created = {2023-01-10T01:44:02.733Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:02.733Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Vascular permeability and angiogenesis underpin neovascular age-related macular degeneration and diabetic retinopathy. While anti-VEGF therapies are widely used clinically, many patients do not respond optimally, or at all, and small-molecule therapies are lacking. Here, we identified a dibenzoxazepinone BT2 that inhibits endothelial cell proliferation, migration, wound repair in vitro, network formation, and angiogenesis in mice bearing Matrigel plugs. BT2 interacts with MEK1 and inhibits ERK phosphorylation and the expression of FosB/ΔFosB, VCAM-1, and many genes involved in proliferation, migration, angiogenesis, and inflammation. BT2 reduced retinal vascular leakage following rat choroidal laser trauma and rabbit intravitreal VEGF-A165 administration. BT2 suppressed retinal CD31, pERK, VCAM-1, and VEGF-A165 expression. BT2 reduced retinal leakage in rats at least as effectively as aflibercept, a first-line therapy for nAMD/DR. BT2 withstands boiling or autoclaving and several months' storage at 22°C. BT2 is a new small-molecule inhibitor of vascular permeability and angiogenesis.},\\n bibtype = {article},\\n author = {Li, Y. and Alhendi, A.M.N. and Yeh, M.-C. and Elahy, M. and Santiago, F.S. and Deshpande, N.P. and Wu, B. and Chan, E. and Inam, S. and Prado-Lourenco, L. and Marcuccio, S.M. and Khachigian, L.M.},\\n doi = {10.1126/sciadv.aaz7815},\\n journal = {Science Advances},\\n number = {31}\\n}\",\"author_short\":[\"Li,  Y.\",\"Alhendi,  A.\",\"Yeh,  M.\",\"Elahy,  M.\",\"Santiago,  F.\",\"Deshpande,  N.\",\"Wu,  B.\",\"Chan,  E.\",\"Inam,  S.\",\"Prado-Lourenco,  L.\",\"Marcuccio,  S.\",\"Khachigian,  L.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"li-alhendi-yeh-elahy-santiago-deshpande-wu-chan-etal-thermostablesmallmoleculeinhibitorofangiogenesisandvascularpermeabilitythatsuppressesaperkfosbfosbvcam1axis-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The Nucleosome Remodeling and Deacetylase Complex Has an Asymmetric, Dynamic, and Modular Architecture\",\"type\":\"article\",\"year\":\"2020\",\"volume\":\"33\",\"id\":\"a494d066-7bf4-3d91-b39e-2c81a3c17fb6\",\"created\":\"2023-01-10T01:44:03.714Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:03.714Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Low et al. examine the architecture of the nucleosome remodeling and deacetylase complex. They define its stoichiometry, use cross-linking mass spectrometry to define subunit locations, and use electron microscopy to reveal large-scale dynamics. They also demonstrate that PWWP2A competes with MBD3 to sequester the HDAC-MTA-RBBP module from NuRD.SCOPUS_ABS_SEPARATORThe nucleosome remodeling and deacetylase (NuRD) complex is essential for metazoan development but has been refractory to biochemical analysis. We present an integrated analysis of the native mammalian NuRD complex, combining quantitative mass spectrometry, cross-linking, protein biochemistry, and electron microscopy to define the architecture of the complex. NuRD is built from a 2:2:4 (MTA, HDAC, and RBBP) deacetylase module and a 1:1:1 (MBD, GATAD2, and Chromodomain-Helicase-DNA-binding [CHD]) remodeling module, and the complex displays considerable structural dynamics. The enigmatic GATAD2 controls the asymmetry of the complex and directly recruits the CHD remodeler. The MTA-MBD interaction acts as a point of functional switching, with the transcriptional regulator PWWP2A competing with MBD for binding to the MTA-HDAC-RBBP subcomplex. Overall, our data address the long-running controversy over NuRD stoichiometry, provide imaging of the mammalian NuRD complex, and establish the biochemical mechanism by which PWWP2A can regulate NuRD composition.\",\"bibtype\":\"article\",\"author\":\"Low, J.K.K. and Silva, A.P.G. and Sharifi Tabar, M. and Torrado, M. and Webb, S.R. and Parker, B.L. and Sana, M. and Smits, C. and Schmidberger, J.W. and Brillault, L. and Landsberg, M.J. and Mackay, J.P.\",\"doi\":\"10.1016/j.celrep.2020.108450\",\"journal\":\"Cell Reports\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {The Nucleosome Remodeling and Deacetylase Complex Has an Asymmetric, Dynamic, and Modular Architecture},\\n type = {article},\\n year = {2020},\\n volume = {33},\\n id = {a494d066-7bf4-3d91-b39e-2c81a3c17fb6},\\n created = {2023-01-10T01:44:03.714Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:03.714Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Low et al. examine the architecture of the nucleosome remodeling and deacetylase complex. They define its stoichiometry, use cross-linking mass spectrometry to define subunit locations, and use electron microscopy to reveal large-scale dynamics. They also demonstrate that PWWP2A competes with MBD3 to sequester the HDAC-MTA-RBBP module from NuRD.SCOPUS_ABS_SEPARATORThe nucleosome remodeling and deacetylase (NuRD) complex is essential for metazoan development but has been refractory to biochemical analysis. We present an integrated analysis of the native mammalian NuRD complex, combining quantitative mass spectrometry, cross-linking, protein biochemistry, and electron microscopy to define the architecture of the complex. NuRD is built from a 2:2:4 (MTA, HDAC, and RBBP) deacetylase module and a 1:1:1 (MBD, GATAD2, and Chromodomain-Helicase-DNA-binding [CHD]) remodeling module, and the complex displays considerable structural dynamics. The enigmatic GATAD2 controls the asymmetry of the complex and directly recruits the CHD remodeler. The MTA-MBD interaction acts as a point of functional switching, with the transcriptional regulator PWWP2A competing with MBD for binding to the MTA-HDAC-RBBP subcomplex. Overall, our data address the long-running controversy over NuRD stoichiometry, provide imaging of the mammalian NuRD complex, and establish the biochemical mechanism by which PWWP2A can regulate NuRD composition.},\\n bibtype = {article},\\n author = {Low, J.K.K. and Silva, A.P.G. and Sharifi Tabar, M. and Torrado, M. and Webb, S.R. and Parker, B.L. and Sana, M. and Smits, C. and Schmidberger, J.W. and Brillault, L. and Landsberg, M.J. and Mackay, J.P.},\\n doi = {10.1016/j.celrep.2020.108450},\\n journal = {Cell Reports},\\n number = {9}\\n}\",\"author_short\":[\"Low,  J.\",\"Silva,  A.\",\"Sharifi Tabar,  M.\",\"Torrado,  M.\",\"Webb,  S.\",\"Parker,  B.\",\"Sana,  M.\",\"Smits,  C.\",\"Schmidberger,  J.\",\"Brillault,  L.\",\"Landsberg,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"low-silva-sharifitabar-torrado-webb-parker-sana-smits-etal-thenucleosomeremodelinganddeacetylasecomplexhasanasymmetricdynamicandmodulararchitecture-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Peppy: A virtual reality environment for exploring the principles of polypeptide structure\",\"type\":\"article\",\"year\":\"2020\",\"pages\":\"157-168\",\"volume\":\"29\",\"id\":\"4e51d150-054e-3635-9211-b383f4822b0e\",\"created\":\"2023-01-10T01:44:04.638Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:04.638Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.\",\"bibtype\":\"article\",\"author\":\"Doak, D.G. and Denyer, G.S. and Gerrard, J.A. and Mackay, J.P. and Allison, J.R.\",\"doi\":\"10.1002/pro.3752\",\"journal\":\"Protein Science\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Peppy: A virtual reality environment for exploring the principles of polypeptide structure},\\n type = {article},\\n year = {2020},\\n pages = {157-168},\\n volume = {29},\\n id = {4e51d150-054e-3635-9211-b383f4822b0e},\\n created = {2023-01-10T01:44:04.638Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:04.638Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.},\\n bibtype = {article},\\n author = {Doak, D.G. and Denyer, G.S. and Gerrard, J.A. and Mackay, J.P. and Allison, J.R.},\\n doi = {10.1002/pro.3752},\\n journal = {Protein Science},\\n number = {1}\\n}\",\"author_short\":[\"Doak,  D.\",\"Denyer,  G.\",\"Gerrard,  J.\",\"Mackay,  J.\",\"Allison,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Correction: GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder (Genetics in Medicine, (2020), 10.1038/s41436-019-0747-z)\",\"type\":\"article\",\"year\":\"2020\",\"pages\":\"822\",\"volume\":\"22\",\"id\":\"7ee81c43-2642-362b-af4d-d6af97c63161\",\"created\":\"2023-01-10T01:44:05.579Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:05.579Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"An amendment to this paper has been published and can be accessed via a link at the top of the paper.\",\"bibtype\":\"article\",\"author\":\"Shieh, C. and Jones, N. and Vanle, B. and Au, M. and Huang, A.Y. and Silva, A.P.G. and Lee, H. and Douine, E.D. and Otero, M.G. and Choi, A. and Mackay, J.P. and Pierson, T.M.\",\"doi\":\"10.1038/s41436-020-0760-2\",\"journal\":\"Genetics in Medicine\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {Correction: GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder (Genetics in Medicine, (2020), 10.1038/s41436-019-0747-z)},\\n type = {article},\\n year = {2020},\\n pages = {822},\\n volume = {22},\\n id = {7ee81c43-2642-362b-af4d-d6af97c63161},\\n created = {2023-01-10T01:44:05.579Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:05.579Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {An amendment to this paper has been published and can be accessed via a link at the top of the paper.},\\n bibtype = {article},\\n author = {Shieh, C. and Jones, N. and Vanle, B. and Au, M. and Huang, A.Y. and Silva, A.P.G. and Lee, H. and Douine, E.D. and Otero, M.G. and Choi, A. and Mackay, J.P. and Pierson, T.M.},\\n doi = {10.1038/s41436-020-0760-2},\\n journal = {Genetics in Medicine},\\n number = {4}\\n}\",\"author_short\":[\"Shieh,  C.\",\"Jones,  N.\",\"Vanle,  B.\",\"Au,  M.\",\"Huang,  A.\",\"Silva,  A.\",\"Lee,  H.\",\"Douine,  E.\",\"Otero,  M.\",\"Choi,  A.\",\"Mackay,  J.\",\"Pierson,  T.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"shieh-jones-vanle-au-huang-silva-lee-douine-etal-correctiongatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisordergeneticsinmedicine2020101038s414360190747z-2020\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G\",\"type\":\"article\",\"year\":\"2019\",\"pages\":\"699-705\",\"volume\":\"87\",\"id\":\"eef77526-79b2-36df-bdf3-98887024df8f\",\"created\":\"2023-01-10T01:44:06.506Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:06.506Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.\",\"bibtype\":\"article\",\"author\":\"Mohanty, B. and Hanson-Manful, P. and Finn, T.J. and Chambers, C.R. and McKellar, J.L.O. and Macindoe, I. and Helder, S. and Setiyaputra, S. and Zhong, Y. and Mackay, J.P. and Mackay, J.P. and Patrick, W.M.\",\"doi\":\"10.1002/prot.25687\",\"journal\":\"Proteins: Structure, Function and Bioinformatics\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G},\\n type = {article},\\n year = {2019},\\n pages = {699-705},\\n volume = {87},\\n id = {eef77526-79b2-36df-bdf3-98887024df8f},\\n created = {2023-01-10T01:44:06.506Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:06.506Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.},\\n bibtype = {article},\\n author = {Mohanty, B. and Hanson-Manful, P. and Finn, T.J. and Chambers, C.R. and McKellar, J.L.O. and Macindoe, I. and Helder, S. and Setiyaputra, S. and Zhong, Y. and Mackay, J.P. and Mackay, J.P. and Patrick, W.M.},\\n doi = {10.1002/prot.25687},\\n journal = {Proteins: Structure, Function and Bioinformatics},\\n number = {8}\\n}\",\"author_short\":[\"Mohanty,  B.\",\"Hanson-Manful,  P.\",\"Finn,  T.\",\"Chambers,  C.\",\"McKellar,  J.\",\"Macindoe,  I.\",\"Helder,  S.\",\"Setiyaputra,  S.\",\"Zhong,  Y.\",\"Mackay,  J.\",\"Mackay,  J.\",\"Patrick,  W.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The stoichiometry and interactome of the Nucleosome Remodeling and Deacetylase (NuRD) complex are conserved across multiple cell lines\",\"type\":\"article\",\"year\":\"2019\",\"pages\":\"2043-2061\",\"volume\":\"286\",\"id\":\"51b93072-bfe2-37ed-9321-35234e2dd662\",\"created\":\"2023-01-10T01:44:07.415Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:07.415Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The nucleosome remodelling and deacetylase complex (NuRD) is a widely conserved regulator of gene expression. The determination of the subunit composition of the complex and identification of its binding partners are important steps towards understanding its architecture and function. The question of how these properties of the complex vary across different cell types has not been addressed in detail to date. Here, we set up a two-step purification protocol coupled to liquid chromatography-tandem mass spectrometry to assess NuRD composition and interaction partners in three different cancer cell lines, using label-free intensity-based absolute quantification (iBAQ). Our data indicate that the stoichiometry of the NuRD complex is preserved across our three different cancer cell lines. In addition, our interactome data suggest ZNF219 and SLC25A5 as possible interaction partners of the complex. To corroborate this latter finding, in vitro and cell-based pull-down experiments were carried out. These experiments indicated that ZNF219 can interact with RBBP4, GATAD2A/B and chromodomain helicase DNA binding 4, whereas SLC25A5 might interact with MTA2 and GATAD2A.\",\"bibtype\":\"article\",\"author\":\"Sharifi Tabar, M. and Mackay, J.P. and Low, J.K.K.\",\"doi\":\"10.1111/febs.14800\",\"journal\":\"FEBS Journal\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {The stoichiometry and interactome of the Nucleosome Remodeling and Deacetylase (NuRD) complex are conserved across multiple cell lines},\\n type = {article},\\n year = {2019},\\n pages = {2043-2061},\\n volume = {286},\\n id = {51b93072-bfe2-37ed-9321-35234e2dd662},\\n created = {2023-01-10T01:44:07.415Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:07.415Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The nucleosome remodelling and deacetylase complex (NuRD) is a widely conserved regulator of gene expression. The determination of the subunit composition of the complex and identification of its binding partners are important steps towards understanding its architecture and function. The question of how these properties of the complex vary across different cell types has not been addressed in detail to date. Here, we set up a two-step purification protocol coupled to liquid chromatography-tandem mass spectrometry to assess NuRD composition and interaction partners in three different cancer cell lines, using label-free intensity-based absolute quantification (iBAQ). Our data indicate that the stoichiometry of the NuRD complex is preserved across our three different cancer cell lines. In addition, our interactome data suggest ZNF219 and SLC25A5 as possible interaction partners of the complex. To corroborate this latter finding, in vitro and cell-based pull-down experiments were carried out. These experiments indicated that ZNF219 can interact with RBBP4, GATAD2A/B and chromodomain helicase DNA binding 4, whereas SLC25A5 might interact with MTA2 and GATAD2A.},\\n bibtype = {article},\\n author = {Sharifi Tabar, M. and Mackay, J.P. and Low, J.K.K.},\\n doi = {10.1111/febs.14800},\\n journal = {FEBS Journal},\\n number = {11}\\n}\",\"author_short\":[\"Sharifi Tabar,  M.\",\"Mackay,  J.\",\"Low,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"sharifitabar-mackay-low-thestoichiometryandinteractomeofthenucleosomeremodelinganddeacetylasenurdcomplexareconservedacrossmultiplecelllines-2019\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":3,\"html\":\"\"},{\"title\":\"Exploring the suitability of RanBP2-type Zinc Fingers for RNA-binding protein design\",\"type\":\"article\",\"year\":\"2019\",\"volume\":\"9\",\"id\":\"221c72ab-8395-34b9-89e2-2cc6f2d798f7\",\"created\":\"2023-01-10T01:44:08.343Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:08.343Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Transcriptomes consist of several classes of RNA that have wide-ranging but often poorly described functions and the deregulation of which leads to numerous diseases. Engineering of functionalized RNA-binding proteins (RBPs) could therefore have many applications. Our previous studies suggested that the RanBP2-type Zinc Finger (ZF) domain is a suitable scaffold to investigate the design of single-stranded RBPs. In the present work, we have analyzed the natural sequence specificity of various members of the RanBP2-type ZF family and characterized the interaction with their target RNA. Surprisingly, our data showed that natural RanBP2-type ZFs with different RNA-binding residues exhibit a similar sequence specificity and therefore no simple recognition code can be established. Despite this finding, different discriminative abilities were observed within the family. In addition, in order to target a long RNA sequence and therefore gain in specificity, we generated a 6-ZF array by combining ZFs from the RanBP2-type family but also from different families, in an effort to achieve a wider target sequence repertoire. We showed that this chimeric protein recognizes its target sequence (20 nucleotides), both in vitro and in living cells. Altogether, our results indicate that the use of ZFs in RBP design remains attractive even though engineering of specificity changes is challenging.\",\"bibtype\":\"article\",\"author\":\"De Franco, S. and Vandenameele, J. and Brans, A. and Verlaine, O. and Bendak, K. and Damblon, C. and Matagne, A. and Segal, D.J. and Galleni, M. and Mackay, J.P. and Mackay, J.P. and Vandevenne, M.\",\"doi\":\"10.1038/s41598-019-38655-y\",\"journal\":\"Scientific Reports\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Exploring the suitability of RanBP2-type Zinc Fingers for RNA-binding protein design},\\n type = {article},\\n year = {2019},\\n volume = {9},\\n id = {221c72ab-8395-34b9-89e2-2cc6f2d798f7},\\n created = {2023-01-10T01:44:08.343Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:08.343Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Transcriptomes consist of several classes of RNA that have wide-ranging but often poorly described functions and the deregulation of which leads to numerous diseases. Engineering of functionalized RNA-binding proteins (RBPs) could therefore have many applications. Our previous studies suggested that the RanBP2-type Zinc Finger (ZF) domain is a suitable scaffold to investigate the design of single-stranded RBPs. In the present work, we have analyzed the natural sequence specificity of various members of the RanBP2-type ZF family and characterized the interaction with their target RNA. Surprisingly, our data showed that natural RanBP2-type ZFs with different RNA-binding residues exhibit a similar sequence specificity and therefore no simple recognition code can be established. Despite this finding, different discriminative abilities were observed within the family. In addition, in order to target a long RNA sequence and therefore gain in specificity, we generated a 6-ZF array by combining ZFs from the RanBP2-type family but also from different families, in an effort to achieve a wider target sequence repertoire. We showed that this chimeric protein recognizes its target sequence (20 nucleotides), both in vitro and in living cells. Altogether, our results indicate that the use of ZFs in RBP design remains attractive even though engineering of specificity changes is challenging.},\\n bibtype = {article},\\n author = {De Franco, S. and Vandenameele, J. and Brans, A. and Verlaine, O. and Bendak, K. and Damblon, C. and Matagne, A. and Segal, D.J. and Galleni, M. and Mackay, J.P. and Mackay, J.P. and Vandevenne, M.},\\n doi = {10.1038/s41598-019-38655-y},\\n journal = {Scientific Reports},\\n number = {1}\\n}\",\"author_short\":[\"De Franco,  S.\",\"Vandenameele,  J.\",\"Brans,  A.\",\"Verlaine,  O.\",\"Bendak,  K.\",\"Damblon,  C.\",\"Matagne,  A.\",\"Segal,  D.\",\"Galleni,  M.\",\"Mackay,  J.\",\"Mackay,  J.\",\"Vandevenne,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"defranco-vandenameele-brans-verlaine-bendak-damblon-matagne-segal-etal-exploringthesuitabilityofranbp2typezincfingersforrnabindingproteindesign-2019\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":6,\"html\":\"\"},{\"title\":\"The NuRD complex and macrocephaly associated neurodevelopmental disorders\",\"type\":\"article\",\"year\":\"2019\",\"pages\":\"548-556\",\"volume\":\"181\",\"id\":\"03edfe18-463d-3f00-9f0b-586a1515ccc8\",\"created\":\"2023-01-10T01:44:09.267Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:09.267Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.\",\"bibtype\":\"article\",\"author\":\"Pierson, T.M. and Otero, M.G. and Grand, K. and Choi, A. and Graham, J.M. and Young, J.I. and Mackay, J.P.\",\"doi\":\"10.1002/ajmg.c.31752\",\"journal\":\"American Journal of Medical Genetics, Part C: Seminars in Medical Genetics\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {The NuRD complex and macrocephaly associated neurodevelopmental disorders},\\n type = {article},\\n year = {2019},\\n pages = {548-556},\\n volume = {181},\\n id = {03edfe18-463d-3f00-9f0b-586a1515ccc8},\\n created = {2023-01-10T01:44:09.267Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:09.267Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.},\\n bibtype = {article},\\n author = {Pierson, T.M. and Otero, M.G. and Grand, K. and Choi, A. and Graham, J.M. and Young, J.I. and Mackay, J.P.},\\n doi = {10.1002/ajmg.c.31752},\\n journal = {American Journal of Medical Genetics, Part C: Seminars in Medical Genetics},\\n number = {4}\\n}\",\"author_short\":[\"Pierson,  T.\",\"Otero,  M.\",\"Grand,  K.\",\"Choi,  A.\",\"Graham,  J.\",\"Young,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding\",\"type\":\"article\",\"year\":\"2018\",\"pages\":\"44-53\",\"volume\":\"16\",\"id\":\"086665b6-9995-388c-8525-8bf8e6dfb235\",\"created\":\"2023-01-10T01:44:10.193Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:10.193Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.\",\"bibtype\":\"article\",\"author\":\"Mundell, S.J. and Rabbolini, D. and Gabrielli, S. and Chen, Q. and Aungraheeta, R. and Hutchinson, J.L. and Kilo, T. and Mackay, J. and Ward, C.M. and Stevenson, W. and Stevenson, W. and Morel-Kopp, M.-C.\",\"doi\":\"10.1111/jth.13900\",\"journal\":\"Journal of Thrombosis and Haemostasis\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding},\\n type = {article},\\n year = {2018},\\n pages = {44-53},\\n volume = {16},\\n id = {086665b6-9995-388c-8525-8bf8e6dfb235},\\n created = {2023-01-10T01:44:10.193Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:10.193Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.},\\n bibtype = {article},\\n author = {Mundell, S.J. and Rabbolini, D. and Gabrielli, S. and Chen, Q. and Aungraheeta, R. and Hutchinson, J.L. and Kilo, T. and Mackay, J. and Ward, C.M. and Stevenson, W. and Stevenson, W. and Morel-Kopp, M.-C.},\\n doi = {10.1111/jth.13900},\\n journal = {Journal of Thrombosis and Haemostasis},\\n number = {1}\\n}\",\"author_short\":[\"Mundell,  S.\",\"Rabbolini,  D.\",\"Gabrielli,  S.\",\"Chen,  Q.\",\"Aungraheeta,  R.\",\"Hutchinson,  J.\",\"Kilo,  T.\",\"Mackay,  J.\",\"Ward,  C.\",\"Stevenson,  W.\",\"Stevenson,  W.\",\"Morel-Kopp,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana\",\"type\":\"article\",\"year\":\"2018\",\"pages\":\"167-176\",\"volume\":\"8\",\"id\":\"dd057f52-1171-342a-ba86-9f93ba6db989\",\"created\":\"2023-01-10T01:44:11.110Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:11.110Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.\",\"bibtype\":\"article\",\"author\":\"Sani, H.M. and Hamzeh-Mivehroud, M. and Silva, A.P. and Walshe, J.L. and Abolghasem Mohammadi, S. and Rahbar-Shahrouziasl, M. and Abbasi, M. and Jamshidi, O. and Low, J.K.K. and Dastmalchi, S. and Dastmalchi, S. and Mackay, J.P.\",\"doi\":\"10.15171/bi.2018.19\",\"journal\":\"BioImpacts\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana},\\n type = {article},\\n year = {2018},\\n pages = {167-176},\\n volume = {8},\\n id = {dd057f52-1171-342a-ba86-9f93ba6db989},\\n created = {2023-01-10T01:44:11.110Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:11.110Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.},\\n bibtype = {article},\\n author = {Sani, H.M. and Hamzeh-Mivehroud, M. and Silva, A.P. and Walshe, J.L. and Abolghasem Mohammadi, S. and Rahbar-Shahrouziasl, M. and Abbasi, M. and Jamshidi, O. and Low, J.K.K. and Dastmalchi, S. and Dastmalchi, S. and Mackay, J.P.},\\n doi = {10.15171/bi.2018.19},\\n journal = {BioImpacts},\\n number = {3}\\n}\",\"author_short\":[\"Sani,  H.\",\"Hamzeh-Mivehroud,  M.\",\"Silva,  A.\",\"Walshe,  J.\",\"Abolghasem Mohammadi,  S.\",\"Rahbar-Shahrouziasl,  M.\",\"Abbasi,  M.\",\"Jamshidi,  O.\",\"Low,  J.\",\"Dastmalchi,  S.\",\"Dastmalchi,  S.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"sani-hamzehmivehroud-silva-walshe-abolghasemmohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"PWWP2A binds distinct chromatin moieties and interacts with an MTA1-specific core NuRD complex\",\"type\":\"article\",\"year\":\"2018\",\"volume\":\"9\",\"id\":\"3ac8e87c-41a6-3ef1-859c-d226ac51ccaa\",\"created\":\"2023-01-10T01:44:12.035Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:12.035Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Chromatin structure and function is regulated by reader proteins recognizing histone modifications and/or histone variants. We recently identified that PWWP2A tightly binds to H2A.Z-containing nucleosomes and is involved in mitotic progression and cranial–facial development. Here, using in vitro assays, we show that distinct domains of PWWP2A mediate binding to free linker DNA as well as H3K36me3 nucleosomes. In vivo, PWWP2A strongly recognizes H2A.Z-containing regulatory regions and weakly binds H3K36me3-containing gene bodies. Further, PWWP2A binds to an MTA1-specific subcomplex of the NuRD complex (M1HR), which consists solely of MTA1, HDAC1, and RBBP4/7, and excludes CHD, GATAD2 and MBD proteins. Depletion of PWWP2A leads to an increase of acetylation levels on H3K27 as well as H2A.Z, presumably by impaired chromatin recruitment of M1HR. Thus, this study identifies PWWP2A as a complex chromatin-binding protein that serves to direct the deacetylase complex M1HR to H2A.Z-containing chromatin, thereby promoting changes in histone acetylation levels.\",\"bibtype\":\"article\",\"author\":\"Link, S. and Spitzer, R.M.M. and Sana, M. and Torrado, M. and Völker-Albert, M.C. and Keilhauer, E.C. and Burgold, T. and Pünzeler, S. and Low, J.K.K. and Lindström, I. and Bartkuhn, M. and Hake, S.B.\",\"doi\":\"10.1038/s41467-018-06665-5\",\"journal\":\"Nature Communications\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {PWWP2A binds distinct chromatin moieties and interacts with an MTA1-specific core NuRD complex},\\n type = {article},\\n year = {2018},\\n volume = {9},\\n id = {3ac8e87c-41a6-3ef1-859c-d226ac51ccaa},\\n created = {2023-01-10T01:44:12.035Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:12.035Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Chromatin structure and function is regulated by reader proteins recognizing histone modifications and/or histone variants. We recently identified that PWWP2A tightly binds to H2A.Z-containing nucleosomes and is involved in mitotic progression and cranial–facial development. Here, using in vitro assays, we show that distinct domains of PWWP2A mediate binding to free linker DNA as well as H3K36me3 nucleosomes. In vivo, PWWP2A strongly recognizes H2A.Z-containing regulatory regions and weakly binds H3K36me3-containing gene bodies. Further, PWWP2A binds to an MTA1-specific subcomplex of the NuRD complex (M1HR), which consists solely of MTA1, HDAC1, and RBBP4/7, and excludes CHD, GATAD2 and MBD proteins. Depletion of PWWP2A leads to an increase of acetylation levels on H3K27 as well as H2A.Z, presumably by impaired chromatin recruitment of M1HR. Thus, this study identifies PWWP2A as a complex chromatin-binding protein that serves to direct the deacetylase complex M1HR to H2A.Z-containing chromatin, thereby promoting changes in histone acetylation levels.},\\n bibtype = {article},\\n author = {Link, S. and Spitzer, R.M.M. and Sana, M. and Torrado, M. and Völker-Albert, M.C. and Keilhauer, E.C. and Burgold, T. and Pünzeler, S. and Low, J.K.K. and Lindström, I. and Bartkuhn, M. and Hake, S.B.},\\n doi = {10.1038/s41467-018-06665-5},\\n journal = {Nature Communications},\\n number = {1}\\n}\",\"author_short\":[\"Link,  S.\",\"Spitzer,  R.\",\"Sana,  M.\",\"Torrado,  M.\",\"Völker-Albert,  M.\",\"Keilhauer,  E.\",\"Burgold,  T.\",\"Pünzeler,  S.\",\"Low,  J.\",\"Lindström,  I.\",\"Bartkuhn,  M.\",\"Hake,  S.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"link-spitzer-sana-torrado-vlkeralbert-keilhauer-burgold-pnzeler-etal-pwwp2abindsdistinctchromatinmoietiesandinteractswithanmta1specificcorenurdcomplex-2018\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"NMR spectroscopy in the analysis of protein-protein interactions\",\"type\":\"book\",\"year\":\"2018\",\"source\":\"Modern Magnetic Resonance\",\"pages\":\"2099-2132\",\"id\":\"8844e241-63ec-3ea2-bdbe-1d62e7545c91\",\"created\":\"2023-01-10T01:44:12.957Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:12.957Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.\",\"bibtype\":\"book\",\"author\":\"Gell, D.A. and Kwan, A.H. and Mackay, J.P.\",\"doi\":\"10.1007/978-3-319-28388-3_121\",\"bibtex\":\"@book{\\n title = {NMR spectroscopy in the analysis of protein-protein interactions},\\n type = {book},\\n year = {2018},\\n source = {Modern Magnetic Resonance},\\n pages = {2099-2132},\\n id = {8844e241-63ec-3ea2-bdbe-1d62e7545c91},\\n created = {2023-01-10T01:44:12.957Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:12.957Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.},\\n bibtype = {book},\\n author = {Gell, D.A. and Kwan, A.H. and Mackay, J.P.},\\n doi = {10.1007/978-3-319-28388-3_121}\\n}\",\"author_short\":[\"Gell,  D.\",\"Kwan,  A.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gell-kwan-mackay-nmrspectroscopyintheanalysisofproteinproteininteractions-2018\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The BRD3 ET domain recognizes a short peptide motif through a mechanism that is conserved across chromatin remodelers and transcriptional regulators\",\"type\":\"article\",\"year\":\"2018\",\"pages\":\"7160-7175\",\"volume\":\"293\",\"id\":\"312373ac-f639-35cd-8df1-cd3bdb1cbb6f\",\"created\":\"2023-01-10T01:44:13.874Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:13.874Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Members of the bromodomain and extra-terminal domain (BET) family of proteins (bromodomain-containing (BRD) 2, 3, 4, and T) are widely expressed and highly conserved regulators of gene expression in eukaryotes. These proteins have been intimately linked to human disease, and more than a dozen clinical trials are currently underway to test BET-protein inhibitors as modulators of cancer. However, although it is clear that these proteins use their bromodomains to bind both histones and transcription factors bearing acetylated lysine residues, the molecular mechanisms by which BET family proteins regulate gene expression are not well defined. In particular, the functions of the other domains such as the ET domain have been less extensively studied. Here, we examine the properties of the ET domain of BRD3 as a protein/protein interaction module. Using a combination of pulldown and biophysical assays, we demonstrate that BRD3 binds to a range of chromatin-remodeling complexes, including the NuRD, BAF, and INO80 complexes, via a short linear “KIKL” motif in one of the complex subunits. NMR-based structural analysis revealed that, surprisingly, this mode of interaction is shared by the AF9 and ENL transcriptional coregulators that contain an acetyl-lysine-binding YEATS domain and regulate transcriptional elongation. This observation establishes a functional commonality between these two families of cancer-related transcriptional regulators. In summary, our data provide insight into the mechanisms by which BET family proteins might link chromatin acetylation to transcriptional outcomes and uncover an unexpected functional similarity between BET and YEATS family proteins.\",\"bibtype\":\"article\",\"author\":\"Wai, D.C.C. and Szyszka, T.N. and Campbell, A.E. and Kwong, C. and Lorna, E.W.-W. and Silva, A.P.G. and Low, J.K.K. and Kwan, A.H. and Gamsjaeger, R. and Chalmers, J.D. and Blobel, G.A. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.RA117.000678\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"19\",\"bibtex\":\"@article{\\n title = {The BRD3 ET domain recognizes a short peptide motif through a mechanism that is conserved across chromatin remodelers and transcriptional regulators},\\n type = {article},\\n year = {2018},\\n pages = {7160-7175},\\n volume = {293},\\n id = {312373ac-f639-35cd-8df1-cd3bdb1cbb6f},\\n created = {2023-01-10T01:44:13.874Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:13.874Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Members of the bromodomain and extra-terminal domain (BET) family of proteins (bromodomain-containing (BRD) 2, 3, 4, and T) are widely expressed and highly conserved regulators of gene expression in eukaryotes. These proteins have been intimately linked to human disease, and more than a dozen clinical trials are currently underway to test BET-protein inhibitors as modulators of cancer. However, although it is clear that these proteins use their bromodomains to bind both histones and transcription factors bearing acetylated lysine residues, the molecular mechanisms by which BET family proteins regulate gene expression are not well defined. In particular, the functions of the other domains such as the ET domain have been less extensively studied. Here, we examine the properties of the ET domain of BRD3 as a protein/protein interaction module. Using a combination of pulldown and biophysical assays, we demonstrate that BRD3 binds to a range of chromatin-remodeling complexes, including the NuRD, BAF, and INO80 complexes, via a short linear “KIKL” motif in one of the complex subunits. NMR-based structural analysis revealed that, surprisingly, this mode of interaction is shared by the AF9 and ENL transcriptional coregulators that contain an acetyl-lysine-binding YEATS domain and regulate transcriptional elongation. This observation establishes a functional commonality between these two families of cancer-related transcriptional regulators. In summary, our data provide insight into the mechanisms by which BET family proteins might link chromatin acetylation to transcriptional outcomes and uncover an unexpected functional similarity between BET and YEATS family proteins.},\\n bibtype = {article},\\n author = {Wai, D.C.C. and Szyszka, T.N. and Campbell, A.E. and Kwong, C. and Lorna, E.W.-W. and Silva, A.P.G. and Low, J.K.K. and Kwan, A.H. and Gamsjaeger, R. and Chalmers, J.D. and Blobel, G.A. and Mackay, J.P.},\\n doi = {10.1074/jbc.RA117.000678},\\n journal = {Journal of Biological Chemistry},\\n number = {19}\\n}\",\"author_short\":[\"Wai,  D.\",\"Szyszka,  T.\",\"Campbell,  A.\",\"Kwong,  C.\",\"Lorna,  E.\",\"Silva,  A.\",\"Low,  J.\",\"Kwan,  A.\",\"Gamsjaeger,  R.\",\"Chalmers,  J.\",\"Blobel,  G.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wai-szyszka-campbell-kwong-lorna-silva-low-kwan-etal-thebrd3etdomainrecognizesashortpeptidemotifthroughamechanismthatisconservedacrosschromatinremodelersandtranscriptionalregulators-2018\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"A Transcription Factor Addiction in Leukemia Imposed by the MLL Promoter Sequence\",\"type\":\"article\",\"year\":\"2018\",\"pages\":\"970-981.e8\",\"volume\":\"34\",\"id\":\"cdabe452-f0bc-3b9a-a76b-3912d34763ca\",\"created\":\"2023-01-10T01:44:14.809Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:14.809Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The Mixed Lineage Leukemia gene (MLL) is altered in leukemia by chromosomal translocations to produce oncoproteins composed of the MLL N-terminus fused to the C-terminus of a partner protein. Here, we used domain-focused CRISPR screening to identify ZFP64 as an essential transcription factor in MLL-rearranged leukemia. We show that the critical function of ZFP64 in leukemia is to maintain MLL expression via binding to the MLL promoter, which is the most enriched location of ZFP64 occupancy in the human genome. The specificity of ZFP64 for MLL is accounted for by an exceptional density of ZFP64 motifs embedded within the MLL promoter. These findings demonstrate how a sequence anomaly of an oncogene promoter can impose a transcriptional addiction in cancer.SCOPUS_ABS_SEPARATORLu et al. show that MLL-rearranged leukemia is addicted to the transcription factor ZFP64 due to direct regulation of MLL by ZFP64. The MLL promoter has an unusually high number of ZFP64 binding motifs and is the most enriched location of ZFP64 occupancy in the human genome.\",\"bibtype\":\"article\",\"author\":\"Lu, B. and Klingbeil, O. and Tarumoto, Y. and Somerville, T.D.D. and Huang, Y.-H. and Wei, Y. and Wai, D.C. and Low, J.K.K. and Milazzo, J.P. and Wu, X.S. and Shi, J. and Vakoc, C.R.\",\"doi\":\"10.1016/j.ccell.2018.10.015\",\"journal\":\"Cancer Cell\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {A Transcription Factor Addiction in Leukemia Imposed by the MLL Promoter Sequence},\\n type = {article},\\n year = {2018},\\n pages = {970-981.e8},\\n volume = {34},\\n id = {cdabe452-f0bc-3b9a-a76b-3912d34763ca},\\n created = {2023-01-10T01:44:14.809Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:14.809Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The Mixed Lineage Leukemia gene (MLL) is altered in leukemia by chromosomal translocations to produce oncoproteins composed of the MLL N-terminus fused to the C-terminus of a partner protein. Here, we used domain-focused CRISPR screening to identify ZFP64 as an essential transcription factor in MLL-rearranged leukemia. We show that the critical function of ZFP64 in leukemia is to maintain MLL expression via binding to the MLL promoter, which is the most enriched location of ZFP64 occupancy in the human genome. The specificity of ZFP64 for MLL is accounted for by an exceptional density of ZFP64 motifs embedded within the MLL promoter. These findings demonstrate how a sequence anomaly of an oncogene promoter can impose a transcriptional addiction in cancer.SCOPUS_ABS_SEPARATORLu et al. show that MLL-rearranged leukemia is addicted to the transcription factor ZFP64 due to direct regulation of MLL by ZFP64. The MLL promoter has an unusually high number of ZFP64 binding motifs and is the most enriched location of ZFP64 occupancy in the human genome.},\\n bibtype = {article},\\n author = {Lu, B. and Klingbeil, O. and Tarumoto, Y. and Somerville, T.D.D. and Huang, Y.-H. and Wei, Y. and Wai, D.C. and Low, J.K.K. and Milazzo, J.P. and Wu, X.S. and Shi, J. and Vakoc, C.R.},\\n doi = {10.1016/j.ccell.2018.10.015},\\n journal = {Cancer Cell},\\n number = {6}\\n}\",\"author_short\":[\"Lu,  B.\",\"Klingbeil,  O.\",\"Tarumoto,  Y.\",\"Somerville,  T.\",\"Huang,  Y.\",\"Wei,  Y.\",\"Wai,  D.\",\"Low,  J.\",\"Milazzo,  J.\",\"Wu,  X.\",\"Shi,  J.\",\"Vakoc,  C.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"lu-klingbeil-tarumoto-somerville-huang-wei-wai-low-etal-atranscriptionfactoraddictioninleukemiaimposedbythemllpromotersequence-2018\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P\",\"type\":\"article\",\"year\":\"2018\",\"pages\":\"1196-1209\",\"volume\":\"19\",\"id\":\"baa2b413-e43a-312a-9a1b-c88fce658272\",\"created\":\"2023-01-10T01:44:15.759Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:15.759Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.\",\"bibtype\":\"article\",\"author\":\"Zhang, X. and Farah, N. and Rolston, L. and Ericsson, D.J. and Catanzariti, A.-M. and Bernoux, M. and Ve, T. and Bendak, K. and Chen, C. and Mackay, J.P. and Jones, D.A. and Kobe, B.\",\"doi\":\"10.1111/mpp.12597\",\"journal\":\"Molecular Plant Pathology\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P},\\n type = {article},\\n year = {2018},\\n pages = {1196-1209},\\n volume = {19},\\n id = {baa2b413-e43a-312a-9a1b-c88fce658272},\\n created = {2023-01-10T01:44:15.759Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:15.759Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.},\\n bibtype = {article},\\n author = {Zhang, X. and Farah, N. and Rolston, L. and Ericsson, D.J. and Catanzariti, A.-M. and Bernoux, M. and Ve, T. and Bendak, K. and Chen, C. and Mackay, J.P. and Jones, D.A. and Kobe, B.},\\n doi = {10.1111/mpp.12597},\\n journal = {Molecular Plant Pathology},\\n number = {5}\\n}\",\"author_short\":[\"Zhang,  X.\",\"Farah,  N.\",\"Rolston,  L.\",\"Ericsson,  D.\",\"Catanzariti,  A.\",\"Bernoux,  M.\",\"Ve,  T.\",\"Bendak,  K.\",\"Chen,  C.\",\"Mackay,  J.\",\"Jones,  D.\",\"Kobe,  B.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex\",\"type\":\"article\",\"year\":\"2017\",\"pages\":\"4216-4232\",\"volume\":\"284\",\"id\":\"125673fd-8a9b-3e40-83fe-795ba612fa34\",\"created\":\"2023-01-10T01:44:16.686Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:16.686Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.\",\"bibtype\":\"article\",\"author\":\"Torrado, M. and Low, J.K.K. and Silva, A.P.G. and Schmidberger, J.W. and Sana, M. and Sharifi Tabar, M. and Isilak, M.E. and Winning, C.S. and Kwong, C. and Bedward, M.J. and Shepherd, N.E. and Mackay, J.P.\",\"doi\":\"10.1111/febs.14301\",\"journal\":\"FEBS Journal\",\"number\":\"24\",\"bibtex\":\"@article{\\n title = {Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex},\\n type = {article},\\n year = {2017},\\n pages = {4216-4232},\\n volume = {284},\\n id = {125673fd-8a9b-3e40-83fe-795ba612fa34},\\n created = {2023-01-10T01:44:16.686Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:16.686Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.},\\n bibtype = {article},\\n author = {Torrado, M. and Low, J.K.K. and Silva, A.P.G. and Schmidberger, J.W. and Sana, M. and Sharifi Tabar, M. and Isilak, M.E. and Winning, C.S. and Kwong, C. and Bedward, M.J. and Shepherd, N.E. and Mackay, J.P.},\\n doi = {10.1111/febs.14301},\\n journal = {FEBS Journal},\\n number = {24}\\n}\",\"author_short\":[\"Torrado,  M.\",\"Low,  J.\",\"Silva,  A.\",\"Schmidberger,  J.\",\"Sana,  M.\",\"Sharifi Tabar,  M.\",\"Isilak,  M.\",\"Winning,  C.\",\"Kwong,  C.\",\"Bedward,  M.\",\"Shepherd,  N.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean\",\"type\":\"article\",\"year\":\"2017\",\"pages\":\"1347-1355.e7\",\"volume\":\"24\",\"id\":\"f5f29eda-8c79-33fb-a24e-f144305413fb\",\"created\":\"2023-01-10T01:44:17.671Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:17.671Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.\",\"bibtype\":\"article\",\"author\":\"Corcilius, L. and Hastwell, A.H. and Zhang, M. and Williams, J. and Mackay, J.P. and Gresshoff, P.M. and Ferguson, B.J. and Payne, R.J.\",\"doi\":\"10.1016/j.chembiol.2017.08.014\",\"journal\":\"Cell Chemical Biology\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean},\\n type = {article},\\n year = {2017},\\n pages = {1347-1355.e7},\\n volume = {24},\\n id = {f5f29eda-8c79-33fb-a24e-f144305413fb},\\n created = {2023-01-10T01:44:17.671Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:17.671Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.},\\n bibtype = {article},\\n author = {Corcilius, L. and Hastwell, A.H. and Zhang, M. and Williams, J. and Mackay, J.P. and Gresshoff, P.M. and Ferguson, B.J. and Payne, R.J.},\\n doi = {10.1016/j.chembiol.2017.08.014},\\n journal = {Cell Chemical Biology},\\n number = {11}\\n}\",\"author_short\":[\"Corcilius,  L.\",\"Hastwell,  A.\",\"Zhang,  M.\",\"Williams,  J.\",\"Mackay,  J.\",\"Gresshoff,  P.\",\"Ferguson,  B.\",\"Payne,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"DCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression\",\"type\":\"article\",\"year\":\"2017\",\"pages\":\"9901-9916\",\"volume\":\"45\",\"id\":\"5eedcada-99ed-3068-9c49-72ae47c1066a\",\"created\":\"2023-01-10T01:44:18.596Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:18.596Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Distinct epigenomic profiles of histone marks have been associated with gene expression, but questions regarding the causal relationship remain. Here we investigated the activity of a broad collection of genomically targeted epigenetic regulators that could write epigenetic marks associated with a repressed chromatin state (G9A, SUV39H1, Krüppel-associated box (KRAB), DNMT3A as well as the first targetable versions of Ezh2 and Friend of GATA-1 (FOG1)). dCas9 fusions produced target gene repression over a range of 0- to 10-fold that varied by locus and cell type. dCpf1 fusions were unable to repress gene expression. The most persistent gene repression required the action of several effector domains; however, KRAB-dCas9 did not contribute to persistence in contrast to previous reports. A 'direct tethering' strategy attaching the Ezh2 methyltransferase enzyme to dCas9, as well as a 'recruitment' strategy attaching the N-terminal 45 residues of FOG1 to dCas9 to recruit the endogenous nucleosome remodeling and deacetylase complex, were both successful in targeted deposition of H3K27me3. Surprisingly, however, repression was not correlated with deposition of either H3K9me3 or H3K27me3. Our results suggest that so-called repressive histone modifications are not sufficient for gene repression. The easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation.\",\"bibtype\":\"article\",\"author\":\"O'Geen, H. and Ren, C. and Nicolet, C.M. and Perez, A.A. and Halmai, J. and Le, V.M. and MacKay, J.P. and Farnham, P.J. and Segal, D.J.\",\"doi\":\"10.1093/nar/gkx578\",\"journal\":\"Nucleic Acids Research\",\"number\":\"17\",\"bibtex\":\"@article{\\n title = {DCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression},\\n type = {article},\\n year = {2017},\\n pages = {9901-9916},\\n volume = {45},\\n id = {5eedcada-99ed-3068-9c49-72ae47c1066a},\\n created = {2023-01-10T01:44:18.596Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:18.596Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Distinct epigenomic profiles of histone marks have been associated with gene expression, but questions regarding the causal relationship remain. Here we investigated the activity of a broad collection of genomically targeted epigenetic regulators that could write epigenetic marks associated with a repressed chromatin state (G9A, SUV39H1, Krüppel-associated box (KRAB), DNMT3A as well as the first targetable versions of Ezh2 and Friend of GATA-1 (FOG1)). dCas9 fusions produced target gene repression over a range of 0- to 10-fold that varied by locus and cell type. dCpf1 fusions were unable to repress gene expression. The most persistent gene repression required the action of several effector domains; however, KRAB-dCas9 did not contribute to persistence in contrast to previous reports. A 'direct tethering' strategy attaching the Ezh2 methyltransferase enzyme to dCas9, as well as a 'recruitment' strategy attaching the N-terminal 45 residues of FOG1 to dCas9 to recruit the endogenous nucleosome remodeling and deacetylase complex, were both successful in targeted deposition of H3K27me3. Surprisingly, however, repression was not correlated with deposition of either H3K9me3 or H3K27me3. Our results suggest that so-called repressive histone modifications are not sufficient for gene repression. The easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation.},\\n bibtype = {article},\\n author = {O'Geen, H. and Ren, C. and Nicolet, C.M. and Perez, A.A. and Halmai, J. and Le, V.M. and MacKay, J.P. and Farnham, P.J. and Segal, D.J.},\\n doi = {10.1093/nar/gkx578},\\n journal = {Nucleic Acids Research},\\n number = {17}\\n}\",\"author_short\":[\"O'Geen,  H.\",\"Ren,  C.\",\"Nicolet,  C.\",\"Perez,  A.\",\"Halmai,  J.\",\"Le,  V.\",\"MacKay,  J.\",\"Farnham,  P.\",\"Segal,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"ogeen-ren-nicolet-perez-halmai-le-mackay-farnham-etal-dcas9basedepigenomeeditingsuggestsacquisitionofhistonemethylationisnotsufficientfortargetgenerepression-2017\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Promiscuous DNA-binding of a mutant zinc finger protein corrupts the transcriptome and diminishes cell viability\",\"type\":\"article\",\"year\":\"2017\",\"pages\":\"1130-1143\",\"volume\":\"45\",\"id\":\"fc32d854-4e6d-3528-9963-88946b3e8087\",\"created\":\"2023-01-10T01:44:19.547Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:19.547Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The rules of engagement between zinc finger transcription factors and DNA have been partly defined by in vitro DNA-binding and structural studies, but less is known about how these rules apply in vivo. Here, we demonstrate how a missense mutation in the second zinc finger of Krüppel-like factor-1 (KLF1) leads to degenerate DNA-binding specificity in vivo, resulting in ectopic transcription and anemia in the Nanmouse model. We employed ChIP-seq and 4sU-RNA-seq to identify aberrant DNA-binding events genome wide and ectopic transcriptional consequences of this binding. We confirmed novel sequence specificity of the mutant recombinant zinc finger domain by performing biophysical measurements ofinvitroDNA-binding affinity. Together, these results shed new light on the mechanisms by which missense mutations in DNA-binding domains of transcription factors can lead to autosomal dominant diseases.\",\"bibtype\":\"article\",\"author\":\"Gillinder, K.R. and Ilsley, M.D. and Nébor, D. and Sachidanandam, R. and Lajoie, M. and Magor, G.W. and Tallack, M.R. and Bailey, T. and Landsberg, M.J. and Mackay, J.P. and Bieker, J.J. and Perkins, A.C.\",\"doi\":\"10.1093/nar/gkw1014\",\"journal\":\"Nucleic Acids Research\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Promiscuous DNA-binding of a mutant zinc finger protein corrupts the transcriptome and diminishes cell viability},\\n type = {article},\\n year = {2017},\\n pages = {1130-1143},\\n volume = {45},\\n id = {fc32d854-4e6d-3528-9963-88946b3e8087},\\n created = {2023-01-10T01:44:19.547Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:19.547Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The rules of engagement between zinc finger transcription factors and DNA have been partly defined by in vitro DNA-binding and structural studies, but less is known about how these rules apply in vivo. Here, we demonstrate how a missense mutation in the second zinc finger of Krüppel-like factor-1 (KLF1) leads to degenerate DNA-binding specificity in vivo, resulting in ectopic transcription and anemia in the Nanmouse model. We employed ChIP-seq and 4sU-RNA-seq to identify aberrant DNA-binding events genome wide and ectopic transcriptional consequences of this binding. We confirmed novel sequence specificity of the mutant recombinant zinc finger domain by performing biophysical measurements ofinvitroDNA-binding affinity. Together, these results shed new light on the mechanisms by which missense mutations in DNA-binding domains of transcription factors can lead to autosomal dominant diseases.},\\n bibtype = {article},\\n author = {Gillinder, K.R. and Ilsley, M.D. and Nébor, D. and Sachidanandam, R. and Lajoie, M. and Magor, G.W. and Tallack, M.R. and Bailey, T. and Landsberg, M.J. and Mackay, J.P. and Bieker, J.J. and Perkins, A.C.},\\n doi = {10.1093/nar/gkw1014},\\n journal = {Nucleic Acids Research},\\n number = {3}\\n}\",\"author_short\":[\"Gillinder,  K.\",\"Ilsley,  M.\",\"Nébor,  D.\",\"Sachidanandam,  R.\",\"Lajoie,  M.\",\"Magor,  G.\",\"Tallack,  M.\",\"Bailey,  T.\",\"Landsberg,  M.\",\"Mackay,  J.\",\"Bieker,  J.\",\"Perkins,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gillinder-ilsley-nbor-sachidanandam-lajoie-magor-tallack-bailey-etal-promiscuousdnabindingofamutantzincfingerproteincorruptsthetranscriptomeanddiminishescellviability-2017\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":2,\"html\":\"\"},{\"title\":\"IP<inf>3-4</inf> kinase Arg1 regulates cell wall homeostasis and surface architecture to promote clearance of Cryptococcus neoformans infection in a mouse model\",\"type\":\"article\",\"year\":\"2017\",\"pages\":\"1833-1848\",\"volume\":\"8\",\"id\":\"2ce6221b-c29d-38b5-826d-bcfd3df72bba\",\"created\":\"2023-01-10T01:44:20.463Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:20.463Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"We previously identified a series of inositol polyphosphate kinases (IPKs), Arg1, Ipk1, Kcs1 and Asp1, in the opportunistic fungal pathogen Cryptococcus neoformans. Using gene deletion analysis, we characterized Arg1, Ipk1 and Kcs1 and showed that they act sequentially to convert IP3 to PP-IP5 (IP7), a key metabolite promoting stress tolerance, metabolic adaptation and fungal dissemination to the brain. We have now directly characterized the enzymatic activity of Arg1, demonstrating that it is a dual specificity (IP3/IP4) kinase producing IP5. We showed previously that IP5 is further phosphorylated by Ipk1 to produce IP6, which is a substrate for the synthesis of PP-IP5 by Kcs1. Phenotypic comparison of the arg1Δ and kcs1Δ deletion mutants (both PP-IP5-deficient) reveals that arg1Δ has the most deleterious phenotype: while PP-IP5 is essential for metabolic and stress adaptation in both mutant strains, PP-IP5 is dispensable for virulence-associated functions such as capsule production, cell wall organization, and normal N-linked mannosylation of the virulence factor, phospholipase B1, as these phenotypes were defective only in arg1Δ. The more deleterious arg1Δ phenotype correlated with a higher rate of arg1Δ phagocytosis by human peripheral blood monocytes and rapid arg1Δ clearance from lung in a mouse model. This observation is in contrast to kcs1Δ, which we previously reported establishes a chronic, confined lung infection. In summary, we show that Arg1 is the most crucial IPK for cryptococcal virulence, conveying PP-IP5–dependent and novel PP-IP5–independent functions.\",\"bibtype\":\"article\",\"author\":\"Li, C. and Lev, S. and Desmarini, D. and Kaufman-Francis, K. and Saiardi, A. and Silva, A.P.G. and Mackay, J.P. and Thompson, P.E. and Sorrell, T.C. and Djordjevic, J.T.\",\"doi\":\"10.1080/21505594.2017.1385692\",\"journal\":\"Virulence\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {IP<inf>3-4</inf> kinase Arg1 regulates cell wall homeostasis and surface architecture to promote clearance of Cryptococcus neoformans infection in a mouse model},\\n type = {article},\\n year = {2017},\\n pages = {1833-1848},\\n volume = {8},\\n id = {2ce6221b-c29d-38b5-826d-bcfd3df72bba},\\n created = {2023-01-10T01:44:20.463Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:20.463Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {We previously identified a series of inositol polyphosphate kinases (IPKs), Arg1, Ipk1, Kcs1 and Asp1, in the opportunistic fungal pathogen Cryptococcus neoformans. Using gene deletion analysis, we characterized Arg1, Ipk1 and Kcs1 and showed that they act sequentially to convert IP3 to PP-IP5 (IP7), a key metabolite promoting stress tolerance, metabolic adaptation and fungal dissemination to the brain. We have now directly characterized the enzymatic activity of Arg1, demonstrating that it is a dual specificity (IP3/IP4) kinase producing IP5. We showed previously that IP5 is further phosphorylated by Ipk1 to produce IP6, which is a substrate for the synthesis of PP-IP5 by Kcs1. Phenotypic comparison of the arg1Δ and kcs1Δ deletion mutants (both PP-IP5-deficient) reveals that arg1Δ has the most deleterious phenotype: while PP-IP5 is essential for metabolic and stress adaptation in both mutant strains, PP-IP5 is dispensable for virulence-associated functions such as capsule production, cell wall organization, and normal N-linked mannosylation of the virulence factor, phospholipase B1, as these phenotypes were defective only in arg1Δ. The more deleterious arg1Δ phenotype correlated with a higher rate of arg1Δ phagocytosis by human peripheral blood monocytes and rapid arg1Δ clearance from lung in a mouse model. This observation is in contrast to kcs1Δ, which we previously reported establishes a chronic, confined lung infection. In summary, we show that Arg1 is the most crucial IPK for cryptococcal virulence, conveying PP-IP5–dependent and novel PP-IP5–independent functions.},\\n bibtype = {article},\\n author = {Li, C. and Lev, S. and Desmarini, D. and Kaufman-Francis, K. and Saiardi, A. and Silva, A.P.G. and Mackay, J.P. and Thompson, P.E. and Sorrell, T.C. and Djordjevic, J.T.},\\n doi = {10.1080/21505594.2017.1385692},\\n journal = {Virulence},\\n number = {8}\\n}\",\"author_short\":[\"Li,  C.\",\"Lev,  S.\",\"Desmarini,  D.\",\"Kaufman-Francis,  K.\",\"Saiardi,  A.\",\"Silva,  A.\",\"Mackay,  J.\",\"Thompson,  P.\",\"Sorrell,  T.\",\"Djordjevic,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"li-lev-desmarini-kaufmanfrancis-saiardi-silva-mackay-thompson-etal-ipinf34infkinasearg1regulatescellwallhomeostasisandsurfacearchitecturetopromoteclearanceofcryptococcusneoformansinfectioninamousemodel-2017\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Whaddaya Know: A Guide to Uncertainty and Subjectivity in Structural Biology\",\"type\":\"article\",\"year\":\"2017\",\"pages\":\"155-167\",\"volume\":\"42\",\"id\":\"cea96057-95a9-35d5-851d-39b4311c3896\",\"created\":\"2023-01-10T01:44:21.385Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:21.385Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The methods of structural biology, while powerful, are technically complex. Although the Protein Data Bank (PDB) provides a repository that allows anyone to download any structure, many users would not appreciate the caveats that should be considered when examining a structure. Here, we describe several key uncertainties associated with the application of X-ray crystallography, NMR spectroscopy, single-particle electron microscopy (SPEM), and small-angle scattering (SAS) to biological macromolecules. The take-home message is that structures are not absolute truths – they are models that fit the experimental data and therefore have uncertainty and subjectivity associated with them. These uncertainties must be appreciated – careful reading of the associated paper, and any validation report provided by the structure database, is highly recommended.\",\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Landsberg, M.J. and Whitten, A.E. and Bond, C.S.\",\"doi\":\"10.1016/j.tibs.2016.11.002\",\"journal\":\"Trends in Biochemical Sciences\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Whaddaya Know: A Guide to Uncertainty and Subjectivity in Structural Biology},\\n type = {article},\\n year = {2017},\\n pages = {155-167},\\n volume = {42},\\n id = {cea96057-95a9-35d5-851d-39b4311c3896},\\n created = {2023-01-10T01:44:21.385Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:21.385Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The methods of structural biology, while powerful, are technically complex. Although the Protein Data Bank (PDB) provides a repository that allows anyone to download any structure, many users would not appreciate the caveats that should be considered when examining a structure. Here, we describe several key uncertainties associated with the application of X-ray crystallography, NMR spectroscopy, single-particle electron microscopy (SPEM), and small-angle scattering (SAS) to biological macromolecules. The take-home message is that structures are not absolute truths – they are models that fit the experimental data and therefore have uncertainty and subjectivity associated with them. These uncertainties must be appreciated – careful reading of the associated paper, and any validation report provided by the structure database, is highly recommended.},\\n bibtype = {article},\\n author = {Mackay, J.P. and Landsberg, M.J. and Whitten, A.E. and Bond, C.S.},\\n doi = {10.1016/j.tibs.2016.11.002},\\n journal = {Trends in Biochemical Sciences},\\n number = {2}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Landsberg,  M.\",\"Whitten,  A.\",\"Bond,  C.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-landsberg-whitten-bond-whaddayaknowaguidetouncertaintyandsubjectivityinstructuralbiology-2017\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"4298-4314\",\"volume\":\"428\",\"id\":\"77987d3c-291d-39e9-97bc-2d692f6cd0c0\",\"created\":\"2023-01-10T01:44:22.294Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:22.294Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.\",\"bibtype\":\"article\",\"author\":\"Mohanty, B. and Helder, S. and Silva, A.P.G. and Mackay, J.P. and Ryan, D.P.\",\"doi\":\"10.1016/j.jmb.2016.08.028\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"21\",\"bibtex\":\"@article{\\n title = {The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain},\\n type = {article},\\n year = {2016},\\n pages = {4298-4314},\\n volume = {428},\\n id = {77987d3c-291d-39e9-97bc-2d692f6cd0c0},\\n created = {2023-01-10T01:44:22.294Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:22.294Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.},\\n bibtype = {article},\\n author = {Mohanty, B. and Helder, S. and Silva, A.P.G. and Mackay, J.P. and Ryan, D.P.},\\n doi = {10.1016/j.jmb.2016.08.028},\\n journal = {Journal of Molecular Biology},\\n number = {21}\\n}\",\"author_short\":[\"Mohanty,  B.\",\"Helder,  S.\",\"Silva,  A.\",\"Mackay,  J.\",\"Ryan,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Site-specific phosphorylation of tau inhibits amyloid-β toxicity in Alzheimer's mice\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"904-908\",\"volume\":\"354\",\"id\":\"569c0c3a-417c-3254-a9b2-e49317df57d6\",\"created\":\"2023-01-10T01:44:23.226Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:23.226Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Amyloid-β (Aβ) toxicity in Alzheimer's disease (AD) is considered to be mediated by phosphorylated tau protein. In contrast, we found that, at least in early disease, site-specific phosphorylation of tau inhibited Aβ toxicity. This specific tau phosphorylation was mediated by the neuronal p38 mitogen-activated protein kinase p38γ and interfered with postsynaptic excitotoxic signaling complexes engaged by Aβ. Accordingly, depletion of p38γ exacerbated neuronal circuit aberrations, cognitive deficits, and premature lethality in a mousemodel of AD, whereas increasing the activity of p38γ abolished these deficits. Furthermore, mimicking site-specific tau phosphorylation alleviated Aβ-induced neuronal death and offered protection from excitotoxicity. Our work provides insights into postsynaptic processes in AD pathogenesis and challenges a purely pathogenic role of tau phosphorylation in neuronal toxicity.\",\"bibtype\":\"article\",\"author\":\"Ittner, A. and Chua, S.W. and Bertz, J. and Volkerling, A. and Van Der Hoven, J. and Gladbach, A. and Przybyla, M. and Bi, M. and Van Hummel, A. and Stevens, C.H. and Ke, Y.D. and Ittner, L.M.\",\"doi\":\"10.1126/science.aah6205\",\"journal\":\"Science\",\"number\":\"6314\",\"bibtex\":\"@article{\\n title = {Site-specific phosphorylation of tau inhibits amyloid-β toxicity in Alzheimer's mice},\\n type = {article},\\n year = {2016},\\n pages = {904-908},\\n volume = {354},\\n id = {569c0c3a-417c-3254-a9b2-e49317df57d6},\\n created = {2023-01-10T01:44:23.226Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:23.226Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Amyloid-β (Aβ) toxicity in Alzheimer's disease (AD) is considered to be mediated by phosphorylated tau protein. In contrast, we found that, at least in early disease, site-specific phosphorylation of tau inhibited Aβ toxicity. This specific tau phosphorylation was mediated by the neuronal p38 mitogen-activated protein kinase p38γ and interfered with postsynaptic excitotoxic signaling complexes engaged by Aβ. Accordingly, depletion of p38γ exacerbated neuronal circuit aberrations, cognitive deficits, and premature lethality in a mousemodel of AD, whereas increasing the activity of p38γ abolished these deficits. Furthermore, mimicking site-specific tau phosphorylation alleviated Aβ-induced neuronal death and offered protection from excitotoxicity. Our work provides insights into postsynaptic processes in AD pathogenesis and challenges a purely pathogenic role of tau phosphorylation in neuronal toxicity.},\\n bibtype = {article},\\n author = {Ittner, A. and Chua, S.W. and Bertz, J. and Volkerling, A. and Van Der Hoven, J. and Gladbach, A. and Przybyla, M. and Bi, M. and Van Hummel, A. and Stevens, C.H. and Ke, Y.D. and Ittner, L.M.},\\n doi = {10.1126/science.aah6205},\\n journal = {Science},\\n number = {6314}\\n}\",\"author_short\":[\"Ittner,  A.\",\"Chua,  S.\",\"Bertz,  J.\",\"Volkerling,  A.\",\"Van Der Hoven,  J.\",\"Gladbach,  A.\",\"Przybyla,  M.\",\"Bi,  M.\",\"Van Hummel,  A.\",\"Stevens,  C.\",\"Ke,  Y.\",\"Ittner,  L.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"ittner-chua-bertz-volkerling-vanderhoven-gladbach-przybyla-bi-etal-sitespecificphosphorylationoftauinhibitsamyloidtoxicityinalzheimersmice-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The zinc fingers of YY1 bind single-stranded RNA with low sequence specificity\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"9153-9165\",\"volume\":\"44\",\"id\":\"448ec1cd-9b90-31ca-8df8-2d37eb36ba76\",\"created\":\"2023-01-10T01:44:24.149Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:24.149Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Classical zinc fingers (ZFs) are traditionally considered to act as sequence-specific DNA-binding domains. More recently, classical ZFs have been recognised as potential RNA-binding modules, raising the intriguing possibility that classical-ZF transcription factors are involved in post-transcriptional gene regulation via direct RNA binding. To date, however, only one classical ZF-RNA complex, that involving TFIIIA, has been structurally characterised. Yin Yang-1 (YY1) is a multi-functional transcription factor involved in many regulatory processes, and binds DNA via four classical ZFs. Recent evidence suggests that YY1 also interacts with RNA, but the molecular nature of the interaction remains unknown. In the present work, we directly assess the ability of YY1 to bind RNA using in vitro assays. Systematic Evolution of Ligands by EXponential enrichment (SELEX) was used to identify preferred RNA sequences bound by the YY1 ZFs from a randomised library over multiple rounds of selection. However, a strong motif was not consistently recovered, suggesting that the RNA sequence selectivity of these domains is modest. YY1 ZF residues involved in binding to single-stranded RNA were identified by NMR spectroscopy and found to be largely distinct from the set of residues involved in DNA binding, suggesting that interactions between YY1 and ssRNA constitute a separate mode of nucleic acid binding. Our data are consistent with recent reports that YY1 can bind to RNA in a low-specificity, yet physiologically relevant manner.\",\"bibtype\":\"article\",\"author\":\"Wai, D.C.C. and Shihab, M. and Low, J.K.K. and Mackay, J.P.\",\"doi\":\"10.1093/nar/gkw590\",\"journal\":\"Nucleic Acids Research\",\"number\":\"19\",\"bibtex\":\"@article{\\n title = {The zinc fingers of YY1 bind single-stranded RNA with low sequence specificity},\\n type = {article},\\n year = {2016},\\n pages = {9153-9165},\\n volume = {44},\\n id = {448ec1cd-9b90-31ca-8df8-2d37eb36ba76},\\n created = {2023-01-10T01:44:24.149Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:24.149Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Classical zinc fingers (ZFs) are traditionally considered to act as sequence-specific DNA-binding domains. More recently, classical ZFs have been recognised as potential RNA-binding modules, raising the intriguing possibility that classical-ZF transcription factors are involved in post-transcriptional gene regulation via direct RNA binding. To date, however, only one classical ZF-RNA complex, that involving TFIIIA, has been structurally characterised. Yin Yang-1 (YY1) is a multi-functional transcription factor involved in many regulatory processes, and binds DNA via four classical ZFs. Recent evidence suggests that YY1 also interacts with RNA, but the molecular nature of the interaction remains unknown. In the present work, we directly assess the ability of YY1 to bind RNA using in vitro assays. Systematic Evolution of Ligands by EXponential enrichment (SELEX) was used to identify preferred RNA sequences bound by the YY1 ZFs from a randomised library over multiple rounds of selection. However, a strong motif was not consistently recovered, suggesting that the RNA sequence selectivity of these domains is modest. YY1 ZF residues involved in binding to single-stranded RNA were identified by NMR spectroscopy and found to be largely distinct from the set of residues involved in DNA binding, suggesting that interactions between YY1 and ssRNA constitute a separate mode of nucleic acid binding. Our data are consistent with recent reports that YY1 can bind to RNA in a low-specificity, yet physiologically relevant manner.},\\n bibtype = {article},\\n author = {Wai, D.C.C. and Shihab, M. and Low, J.K.K. and Mackay, J.P.},\\n doi = {10.1093/nar/gkw590},\\n journal = {Nucleic Acids Research},\\n number = {19}\\n}\",\"author_short\":[\"Wai,  D.\",\"Shihab,  M.\",\"Low,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wai-shihab-low-mackay-thezincfingersofyy1bindsinglestrandedrnawithlowsequencespecificity-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Determinants of affinity and specificity in RNA-binding proteins\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"83-91\",\"volume\":\"38\",\"id\":\"61fe4e83-b401-35f1-9433-78ccbe3d486b\",\"created\":\"2023-01-10T01:44:25.084Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:25.084Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Emerging data suggest that the mechanisms by which RNA-binding proteins (RBPs) interact with RNA and the rules governing specificity might be substantially more complex than those underlying their DNA-binding counterparts. Even our knowledge of what constitutes the RNA-bound proteome is contentious; recent studies suggest that 10-30% of RBPs contain no known RNA-binding domain. Adding to this situation is a growing disconnect between the avalanche of identified interactions between proteins and long noncoding RNAs and the absence of biophysical data on these interactions. RNA-protein interactions are also at the centre of what might emerge as one of the biggest shifts in thinking about cell and molecular biology this century, following from recent reports of ribonucleoprotein complexes that drive reversible membrane-free phase separation events within the cell. Unexpectedly, low-complexity motifs are important in the formation of these structures. Here we briefly survey recent advances in our understanding of the specificity of RBPs.\",\"bibtype\":\"article\",\"author\":\"Helder, S. and Blythe, A.J. and Bond, C.S. and Mackay, J.P.\",\"doi\":\"10.1016/j.sbi.2016.05.005\",\"journal\":\"Current Opinion in Structural Biology\",\"bibtex\":\"@article{\\n title = {Determinants of affinity and specificity in RNA-binding proteins},\\n type = {article},\\n year = {2016},\\n pages = {83-91},\\n volume = {38},\\n id = {61fe4e83-b401-35f1-9433-78ccbe3d486b},\\n created = {2023-01-10T01:44:25.084Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:25.084Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Emerging data suggest that the mechanisms by which RNA-binding proteins (RBPs) interact with RNA and the rules governing specificity might be substantially more complex than those underlying their DNA-binding counterparts. Even our knowledge of what constitutes the RNA-bound proteome is contentious; recent studies suggest that 10-30% of RBPs contain no known RNA-binding domain. Adding to this situation is a growing disconnect between the avalanche of identified interactions between proteins and long noncoding RNAs and the absence of biophysical data on these interactions. RNA-protein interactions are also at the centre of what might emerge as one of the biggest shifts in thinking about cell and molecular biology this century, following from recent reports of ribonucleoprotein complexes that drive reversible membrane-free phase separation events within the cell. Unexpectedly, low-complexity motifs are important in the formation of these structures. Here we briefly survey recent advances in our understanding of the specificity of RBPs.},\\n bibtype = {article},\\n author = {Helder, S. and Blythe, A.J. and Bond, C.S. and Mackay, J.P.},\\n doi = {10.1016/j.sbi.2016.05.005},\\n journal = {Current Opinion in Structural Biology}\\n}\",\"author_short\":[\"Helder,  S.\",\"Blythe,  A.\",\"Bond,  C.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"helder-blythe-bond-mackay-determinantsofaffinityandspecificityinrnabindingproteins-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"1472-1482\",\"id\":\"a9a74730-2adb-3cee-87b3-9643769fab42\",\"created\":\"2023-01-10T01:44:26.953Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:26.953Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.\",\"bibtype\":\"article\",\"author\":\"Schmidberger, J.W. and Sharifi Tabar, M. and Torrado, M. and Silva, A.P.G. and Landsberg, M.J. and Brillault, L. and AlQarni, S. and Zeng, Y.C. and Parker, B.L. and Low, J.K.K. and Low, J.K.K. and Mackay, J.P.\",\"doi\":\"10.1002/pro.2943\",\"journal\":\"Protein Science\",\"bibtex\":\"@article{\\n title = {The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7},\\n type = {article},\\n year = {2016},\\n pages = {1472-1482},\\n id = {a9a74730-2adb-3cee-87b3-9643769fab42},\\n created = {2023-01-10T01:44:26.953Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:26.953Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.},\\n bibtype = {article},\\n author = {Schmidberger, J.W. and Sharifi Tabar, M. and Torrado, M. and Silva, A.P.G. and Landsberg, M.J. and Brillault, L. and AlQarni, S. and Zeng, Y.C. and Parker, B.L. and Low, J.K.K. and Low, J.K.K. and Mackay, J.P.},\\n doi = {10.1002/pro.2943},\\n journal = {Protein Science}\\n}\",\"author_short\":[\"Schmidberger,  J.\",\"Sharifi Tabar,  M.\",\"Torrado,  M.\",\"Silva,  A.\",\"Landsberg,  M.\",\"Brillault,  L.\",\"AlQarni,  S.\",\"Zeng,  Y.\",\"Parker,  B.\",\"Low,  J.\",\"Low,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The N-terminal region of chromodomain helicase DNA-binding protein 4 (CHD4) is essential for activity and contains a high mobility group (HMG) box-like-domain that can bind poly(ADP-ribose)\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"924-938\",\"volume\":\"291\",\"id\":\"ecf7907f-b86b-3542-ab23-e2a9a7ff5cc9\",\"created\":\"2023-01-10T01:44:27.879Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:27.879Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Chromodomain Helicase DNA-binding protein 4 (CHD4) is a chromatin-remodeling enzyme that has been reported to regulate DNA-damage responses through its N-terminal region in a poly(ADP-ribose) polymerase-dependent manner. We have identified and determined the structure of a stable domain (CHD4-N) in this N-terminal region. The-fold consists of a four-α-helix bundle with structural similarity to the high mobility group box, a domain that is well known as a DNA binding module. We show that the CHD4-N domain binds with higher affinity to poly(ADP-ribose) than to DNA. We also show that the N-terminal region of CHD4, although not CHD4-N alone, is essential for full nucleosome remodeling activity and is important for localizing CHD4 to sites of DNA damage. Overall, these data build on our understanding of how CHD4-NuRD acts to regulate gene expression and participates in the DNA-damage response.\",\"bibtype\":\"article\",\"author\":\"Silva, A.P.G. and Ryan, D.P. and Galanty, Y. and Low, J.K.K. and Vandevenne, M. and Jackson, S.P. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M115.683227\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {The N-terminal region of chromodomain helicase DNA-binding protein 4 (CHD4) is essential for activity and contains a high mobility group (HMG) box-like-domain that can bind poly(ADP-ribose)},\\n type = {article},\\n year = {2016},\\n pages = {924-938},\\n volume = {291},\\n id = {ecf7907f-b86b-3542-ab23-e2a9a7ff5cc9},\\n created = {2023-01-10T01:44:27.879Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:27.879Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Chromodomain Helicase DNA-binding protein 4 (CHD4) is a chromatin-remodeling enzyme that has been reported to regulate DNA-damage responses through its N-terminal region in a poly(ADP-ribose) polymerase-dependent manner. We have identified and determined the structure of a stable domain (CHD4-N) in this N-terminal region. The-fold consists of a four-α-helix bundle with structural similarity to the high mobility group box, a domain that is well known as a DNA binding module. We show that the CHD4-N domain binds with higher affinity to poly(ADP-ribose) than to DNA. We also show that the N-terminal region of CHD4, although not CHD4-N alone, is essential for full nucleosome remodeling activity and is important for localizing CHD4 to sites of DNA damage. Overall, these data build on our understanding of how CHD4-NuRD acts to regulate gene expression and participates in the DNA-damage response.},\\n bibtype = {article},\\n author = {Silva, A.P.G. and Ryan, D.P. and Galanty, Y. and Low, J.K.K. and Vandevenne, M. and Jackson, S.P. and Mackay, J.P.},\\n doi = {10.1074/jbc.M115.683227},\\n journal = {Journal of Biological Chemistry},\\n number = {2}\\n}\",\"author_short\":[\"Silva,  A.\",\"Ryan,  D.\",\"Galanty,  Y.\",\"Low,  J.\",\"Vandevenne,  M.\",\"Jackson,  S.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"silva-ryan-galanty-low-vandevenne-jackson-mackay-thenterminalregionofchromodomainhelicasednabindingprotein4chd4isessentialforactivityandcontainsahighmobilitygrouphmgboxlikedomainthatcanbindpolyadpribose-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"CHD4 is a peripheral component of the nucleosome remodeling and deacetylase complex\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"15853-15866\",\"volume\":\"291\",\"id\":\"7b57ef93-0acb-3843-a769-694abe573e25\",\"created\":\"2023-01-10T01:44:28.938Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:28.938Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Chromatin remodeling enzymes act to dynamically regulate gene accessibility. In many cases, these enzymes function as large multicomponent complexes that in general comprise a central ATP-dependent Snf2 family helicase that is decorated with a variable number of regulatory subunits. The nucleosome remodeling and deacetylase (NuRD) complex, which is essential for normal development in higher organisms, is one such macromolecular machine. The NuRD complex comprises ∼10 subunits, including the histone deacetylases 1 and 2 (HDAC1 and HDAC2), and is defined by the presence of a CHD family remodeling enzyme, most commonly CHD4 (chromodomain helicase DNA-binding protein 4). The existing paradigm holds that CHD4 acts as the central hub upon which the complex is built. We show here that this paradigm does not, in fact, hold and that CHD4 is a peripheral component of the NuRD complex. A complex lacking CHD4 that has HDAC activity can exist as a stable species. The addition of recombinant CHD4 to this nucleosome deacetylase complex reconstitutes a NuRD complex with nucleosome remodeling activity. These data contribute to our understanding of the architecture of the NuRD complex.\",\"bibtype\":\"article\",\"author\":\"Low, J.K.K. and Webb, S.R. and Silva, A.P.G. and Saathoff, H. and Ryan, D.P. and Torrado, M. and Brofelth, M. and Parker, B.L. and Shepherd, N.E. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M115.707018\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"30\",\"bibtex\":\"@article{\\n title = {CHD4 is a peripheral component of the nucleosome remodeling and deacetylase complex},\\n type = {article},\\n year = {2016},\\n pages = {15853-15866},\\n volume = {291},\\n id = {7b57ef93-0acb-3843-a769-694abe573e25},\\n created = {2023-01-10T01:44:28.938Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:28.938Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Chromatin remodeling enzymes act to dynamically regulate gene accessibility. In many cases, these enzymes function as large multicomponent complexes that in general comprise a central ATP-dependent Snf2 family helicase that is decorated with a variable number of regulatory subunits. The nucleosome remodeling and deacetylase (NuRD) complex, which is essential for normal development in higher organisms, is one such macromolecular machine. The NuRD complex comprises ∼10 subunits, including the histone deacetylases 1 and 2 (HDAC1 and HDAC2), and is defined by the presence of a CHD family remodeling enzyme, most commonly CHD4 (chromodomain helicase DNA-binding protein 4). The existing paradigm holds that CHD4 acts as the central hub upon which the complex is built. We show here that this paradigm does not, in fact, hold and that CHD4 is a peripheral component of the NuRD complex. A complex lacking CHD4 that has HDAC activity can exist as a stable species. The addition of recombinant CHD4 to this nucleosome deacetylase complex reconstitutes a NuRD complex with nucleosome remodeling activity. These data contribute to our understanding of the architecture of the NuRD complex.},\\n bibtype = {article},\\n author = {Low, J.K.K. and Webb, S.R. and Silva, A.P.G. and Saathoff, H. and Ryan, D.P. and Torrado, M. and Brofelth, M. and Parker, B.L. and Shepherd, N.E. and Mackay, J.P.},\\n doi = {10.1074/jbc.M115.707018},\\n journal = {Journal of Biological Chemistry},\\n number = {30}\\n}\",\"author_short\":[\"Low,  J.\",\"Webb,  S.\",\"Silva,  A.\",\"Saathoff,  H.\",\"Ryan,  D.\",\"Torrado,  M.\",\"Brofelth,  M.\",\"Parker,  B.\",\"Shepherd,  N.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"low-webb-silva-saathoff-ryan-torrado-brofelth-parker-etal-chd4isaperipheralcomponentofthenucleosomeremodelinganddeacetylasecomplex-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The binding of syndapin SH3 domain to Dynamin proline-rich domain involves short and long distance elements\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"9411-9424\",\"volume\":\"291\",\"id\":\"34917dfc-3a3c-379e-a481-47c891a62429\",\"created\":\"2023-01-10T01:44:29.922Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:29.922Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Dynamin is a GTPase that mediates vesicle fission during synaptic vesicle endocytosis. Its long C-terminal proline-rich domain contains 13 PXXP motifs, which orchestrate its interactions with multiple proteins. The SH3 domains of syndapin and endophilin bind the PXXP motifs called Site 2 and 3 (Pro-786-Pro-793) at the N-terminal end of the proline-rich domain, whereas the amphiphysin SH3 binds Site 9 (Pro-833-Pro-836) toward the C-terminal end. In some proteins, SH3/peptide interactions also involve short distance elements, which are 5-15 amino acid extensions flanking the central PXXP motif for high affinity binding. Here we found two previously unrecognized elements in the central and the C-terminal end of the dynamin proline-rich domain that account for a significant increase in syndapin binding affinity compared with a previously reported Site 2 and Site 3 PXXP peptide alone. The first new element (Gly-807-Gly-811) is short distance element on the C-terminal side of Site 2 PXXP, which might contact a groove identified under the RT loop of the SH3 domain. The second element (Arg-838-Pro-844) is located about 50 amino acids downstream of Site 2. These two elements provide additional specificity tothe syndapin SH3 domain outsideofthe well described polyproline-binding groove. Thus, the dynamin/syndapin interaction is mediated via a network of multiple contacts outside the core PXXP motif over a previously unrecognized extended region of the proline-rich domain. To our knowledge this is the first example among known SH3 interactions to involve spatially separated and extended long-range elements that combine to provide a higher affinity interaction. 7copy;2016 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Luo, L. and Xue, J. and Kwan, A. and Gamsjaeger, R. and Wielens, J. and Von Kleist, L. and Cubeddu, L. and Guo, Z. and Stow, J.L. and Parker, M.W. and Mackay, J.P. and Robinson, P.J.\",\"doi\":\"10.1074/jbc.M115.703108\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"18\",\"bibtex\":\"@article{\\n title = {The binding of syndapin SH3 domain to Dynamin proline-rich domain involves short and long distance elements},\\n type = {article},\\n year = {2016},\\n pages = {9411-9424},\\n volume = {291},\\n id = {34917dfc-3a3c-379e-a481-47c891a62429},\\n created = {2023-01-10T01:44:29.922Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:29.922Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Dynamin is a GTPase that mediates vesicle fission during synaptic vesicle endocytosis. Its long C-terminal proline-rich domain contains 13 PXXP motifs, which orchestrate its interactions with multiple proteins. The SH3 domains of syndapin and endophilin bind the PXXP motifs called Site 2 and 3 (Pro-786-Pro-793) at the N-terminal end of the proline-rich domain, whereas the amphiphysin SH3 binds Site 9 (Pro-833-Pro-836) toward the C-terminal end. In some proteins, SH3/peptide interactions also involve short distance elements, which are 5-15 amino acid extensions flanking the central PXXP motif for high affinity binding. Here we found two previously unrecognized elements in the central and the C-terminal end of the dynamin proline-rich domain that account for a significant increase in syndapin binding affinity compared with a previously reported Site 2 and Site 3 PXXP peptide alone. The first new element (Gly-807-Gly-811) is short distance element on the C-terminal side of Site 2 PXXP, which might contact a groove identified under the RT loop of the SH3 domain. The second element (Arg-838-Pro-844) is located about 50 amino acids downstream of Site 2. These two elements provide additional specificity tothe syndapin SH3 domain outsideofthe well described polyproline-binding groove. Thus, the dynamin/syndapin interaction is mediated via a network of multiple contacts outside the core PXXP motif over a previously unrecognized extended region of the proline-rich domain. To our knowledge this is the first example among known SH3 interactions to involve spatially separated and extended long-range elements that combine to provide a higher affinity interaction. 7copy;2016 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Luo, L. and Xue, J. and Kwan, A. and Gamsjaeger, R. and Wielens, J. and Von Kleist, L. and Cubeddu, L. and Guo, Z. and Stow, J.L. and Parker, M.W. and Mackay, J.P. and Robinson, P.J.},\\n doi = {10.1074/jbc.M115.703108},\\n journal = {Journal of Biological Chemistry},\\n number = {18}\\n}\",\"author_short\":[\"Luo,  L.\",\"Xue,  J.\",\"Kwan,  A.\",\"Gamsjaeger,  R.\",\"Wielens,  J.\",\"Von Kleist,  L.\",\"Cubeddu,  L.\",\"Guo,  Z.\",\"Stow,  J.\",\"Parker,  M.\",\"Mackay,  J.\",\"Robinson,  P.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"luo-xue-kwan-gamsjaeger-wielens-vonkleist-cubeddu-guo-etal-thebindingofsyndapinsh3domaintodynaminprolinerichdomaininvolvesshortandlongdistanceelements-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of a C-terminal domain of human CHD1\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"31-34\",\"volume\":\"10\",\"id\":\"b856848d-bf8e-37fe-813e-40d403bbd998\",\"created\":\"2023-01-10T01:44:30.848Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:30.848Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Chromatin remodelling proteins are an essential family of eukaryotic proteins. They harness the energy from ATP hydrolysis and apply it to alter chromatin structure in order to regulate all aspects of genome biology. Chromodomain helicase DNA-binding protein 1 (CHD1) is one such remodelling protein that has specialised nucleosome organising abilities and is conserved across eukaryotes. CHD1 possesses a pair of tandem chromodomains that directly precede the core catalytic Snf2 helicase-like domain, and a C-terminal SANT-SLIDE DNA-binding domain. We have identified an additional conserved domain in the C-terminal region of CHD1. Here, we report the backbone and side chain resonance assignments for this domain from human CHD1 at pH 6.5 and 25 °C (BMRB No. 25638).\",\"bibtype\":\"article\",\"author\":\"Mohanty, B. and Silva, A.P.G. and Mackay, J.P. and Ryan, D.P.\",\"doi\":\"10.1007/s12104-015-9631-1\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of a C-terminal domain of human CHD1},\\n type = {article},\\n year = {2016},\\n pages = {31-34},\\n volume = {10},\\n id = {b856848d-bf8e-37fe-813e-40d403bbd998},\\n created = {2023-01-10T01:44:30.848Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:30.848Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Chromatin remodelling proteins are an essential family of eukaryotic proteins. They harness the energy from ATP hydrolysis and apply it to alter chromatin structure in order to regulate all aspects of genome biology. Chromodomain helicase DNA-binding protein 1 (CHD1) is one such remodelling protein that has specialised nucleosome organising abilities and is conserved across eukaryotes. CHD1 possesses a pair of tandem chromodomains that directly precede the core catalytic Snf2 helicase-like domain, and a C-terminal SANT-SLIDE DNA-binding domain. We have identified an additional conserved domain in the C-terminal region of CHD1. Here, we report the backbone and side chain resonance assignments for this domain from human CHD1 at pH 6.5 and 25 °C (BMRB No. 25638).},\\n bibtype = {article},\\n author = {Mohanty, B. and Silva, A.P.G. and Mackay, J.P. and Ryan, D.P.},\\n doi = {10.1007/s12104-015-9631-1},\\n journal = {Biomolecular NMR Assignments},\\n number = {1}\\n}\",\"author_short\":[\"Mohanty,  B.\",\"Silva,  A.\",\"Mackay,  J.\",\"Ryan,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mohanty-silva-mackay-ryan-sup1suphsup13supcandsup15supnresonanceassignmentsofacterminaldomainofhumanchd1-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein\",\"type\":\"article\",\"year\":\"2016\",\"pages\":\"1710-1721\",\"id\":\"7411c703-cf65-3456-a534-495980102c40\",\"created\":\"2023-01-10T01:44:31.837Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:31.837Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.\",\"bibtype\":\"article\",\"author\":\"Blythe, A.J. and Yazar-Klosinski, B. and Webster, M.W. and Chen, E. and Vandevenne, M. and Bendak, K. and Mackay, J.P. and Hartzog, G.A. and Vrielink, A.\",\"doi\":\"10.1002/pro.2976\",\"journal\":\"Protein Science\",\"bibtex\":\"@article{\\n title = {The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein},\\n type = {article},\\n year = {2016},\\n pages = {1710-1721},\\n id = {7411c703-cf65-3456-a534-495980102c40},\\n created = {2023-01-10T01:44:31.837Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:31.837Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.},\\n bibtype = {article},\\n author = {Blythe, A.J. and Yazar-Klosinski, B. and Webster, M.W. and Chen, E. and Vandevenne, M. and Bendak, K. and Mackay, J.P. and Hartzog, G.A. and Vrielink, A.},\\n doi = {10.1002/pro.2976},\\n journal = {Protein Science}\\n}\",\"author_short\":[\"Blythe,  A.\",\"Yazar-Klosinski,  B.\",\"Webster,  M.\",\"Chen,  E.\",\"Vandevenne,  M.\",\"Bendak,  K.\",\"Mackay,  J.\",\"Hartzog,  G.\",\"Vrielink,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Paris-Trousseau thrombocytopenia is phenocopied by the autosomal recessive inheritance of a DNA-binding domain mutation in FLI1\",\"type\":\"article\",\"year\":\"2015\",\"pages\":\"2027-2030\",\"volume\":\"126\",\"id\":\"9411fc17-2fe7-33f7-b1a3-3b0944dfec36\",\"created\":\"2023-01-10T01:44:32.853Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:32.853Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Hemizygous deletion of a variable regionon chromosome 11q containing FLI1 causes an inherited platelet-related bleeding disorder in Paris-Trousseau thrombocytopenia and Jacobsen syndrome. These multisystem disorders are also characterized by heart anomalies, changes in facial structure, and intellectual disability. We have identified a consanguineous family with autosomal recessive inheritance of a bleeding disorder that mimics Paris-Trousseau thrombocytopenia but has no other features of the 11q23 deletion syndrome. Affected individuals inthis family have moderate thrombocytopenia; absent collagen-induced platelet aggregation; and large, fused α-granules in1%to 5% of circulating platelets. This phenotype was caused by a FLI1 homozygous c.970C>T-point mutation that predicts an arginine-to-tryptophan substitution in the conserved ETS DNA-binding domain of FLI1. This mutation caused a transcription defect at the promoter of known FLI1 target genes GP6, GP9, and ITGA2B, as measured by luciferase assay in HEK293 cells, and decreased the expression of these target proteins in affected members of the family as measured by Western blotting of platelet lysates. This kindred suggests abnormalitiesin FLI1 as causative of Paris-Trousseau thrombocytopenia and confirms the important role of FLI1 in normal platelet development.\",\"bibtype\":\"article\",\"author\":\"Stevenson, W.S. and Rabbolini, D.J. and Beutler, L. and Chen, Q. and Gabrielli, S. and Mackay, J.P. and Brighton, T.A. and Ward, C.M. and Morel-Kopp, M.-C.\",\"doi\":\"10.1182/blood-2015-06-650887\",\"journal\":\"Blood\",\"number\":\"17\",\"bibtex\":\"@article{\\n title = {Paris-Trousseau thrombocytopenia is phenocopied by the autosomal recessive inheritance of a DNA-binding domain mutation in FLI1},\\n type = {article},\\n year = {2015},\\n pages = {2027-2030},\\n volume = {126},\\n id = {9411fc17-2fe7-33f7-b1a3-3b0944dfec36},\\n created = {2023-01-10T01:44:32.853Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:32.853Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Hemizygous deletion of a variable regionon chromosome 11q containing FLI1 causes an inherited platelet-related bleeding disorder in Paris-Trousseau thrombocytopenia and Jacobsen syndrome. These multisystem disorders are also characterized by heart anomalies, changes in facial structure, and intellectual disability. We have identified a consanguineous family with autosomal recessive inheritance of a bleeding disorder that mimics Paris-Trousseau thrombocytopenia but has no other features of the 11q23 deletion syndrome. Affected individuals inthis family have moderate thrombocytopenia; absent collagen-induced platelet aggregation; and large, fused α-granules in1%to 5% of circulating platelets. This phenotype was caused by a FLI1 homozygous c.970C>T-point mutation that predicts an arginine-to-tryptophan substitution in the conserved ETS DNA-binding domain of FLI1. This mutation caused a transcription defect at the promoter of known FLI1 target genes GP6, GP9, and ITGA2B, as measured by luciferase assay in HEK293 cells, and decreased the expression of these target proteins in affected members of the family as measured by Western blotting of platelet lysates. This kindred suggests abnormalitiesin FLI1 as causative of Paris-Trousseau thrombocytopenia and confirms the important role of FLI1 in normal platelet development.},\\n bibtype = {article},\\n author = {Stevenson, W.S. and Rabbolini, D.J. and Beutler, L. and Chen, Q. and Gabrielli, S. and Mackay, J.P. and Brighton, T.A. and Ward, C.M. and Morel-Kopp, M.-C.},\\n doi = {10.1182/blood-2015-06-650887},\\n journal = {Blood},\\n number = {17}\\n}\",\"author_short\":[\"Stevenson,  W.\",\"Rabbolini,  D.\",\"Beutler,  L.\",\"Chen,  Q.\",\"Gabrielli,  S.\",\"Mackay,  J.\",\"Brighton,  T.\",\"Ward,  C.\",\"Morel-Kopp,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"stevenson-rabbolini-beutler-chen-gabrielli-mackay-brighton-ward-etal-paristrousseauthrombocytopeniaisphenocopiedbytheautosomalrecessiveinheritanceofadnabindingdomainmutationinfli1-2015\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Site directed nitroxide spin labeling of oligonucleotides for NMR and EPR studies\",\"type\":\"article\",\"year\":\"2015\",\"pages\":\"813-819\",\"volume\":\"71\",\"id\":\"2ba001b8-f1dd-3136-90e2-56100f2c35ff\",\"created\":\"2023-01-10T01:44:33.864Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:33.864Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Nitroxide labels are useful probes of biomolecule structure that can be detected by NMR and EPR spectroscopy. Although many methods exist for labeling oligonucleotides with nitroxides, most require reagents that are expensive or laborious to prepare. A simpler approach is described herein using commercially available phosphorothioate oligonucleotides and 2-(3-iodoacetamidomethyl)-PROXYL. We describe semi-optimized conditions for labeling DNA and RNA oligonucleotides and methodology for purifying and identifying these reagents by MALDI-MS. The nitroxide label showed some propensity to hydrolyze at high temperature and over prolonged periods at room temperature. Nitroxide-labeled DNA oligonucleotides gave characteristic EPR spectra and caused the disappearance of bound protein signals in 15N HSQC spectra consistent with the paramagnetic relaxation enhancement (PRE) effect.\",\"bibtype\":\"article\",\"author\":\"Shepherd, N.E. and Gamsjaeger, R. and Vandevenne, M. and Cubeddu, L. and Mackay, J.P.\",\"doi\":\"10.1016/j.tet.2014.12.056\",\"journal\":\"Tetrahedron\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Site directed nitroxide spin labeling of oligonucleotides for NMR and EPR studies},\\n type = {article},\\n year = {2015},\\n pages = {813-819},\\n volume = {71},\\n id = {2ba001b8-f1dd-3136-90e2-56100f2c35ff},\\n created = {2023-01-10T01:44:33.864Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:33.864Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Nitroxide labels are useful probes of biomolecule structure that can be detected by NMR and EPR spectroscopy. Although many methods exist for labeling oligonucleotides with nitroxides, most require reagents that are expensive or laborious to prepare. A simpler approach is described herein using commercially available phosphorothioate oligonucleotides and 2-(3-iodoacetamidomethyl)-PROXYL. We describe semi-optimized conditions for labeling DNA and RNA oligonucleotides and methodology for purifying and identifying these reagents by MALDI-MS. The nitroxide label showed some propensity to hydrolyze at high temperature and over prolonged periods at room temperature. Nitroxide-labeled DNA oligonucleotides gave characteristic EPR spectra and caused the disappearance of bound protein signals in 15N HSQC spectra consistent with the paramagnetic relaxation enhancement (PRE) effect.},\\n bibtype = {article},\\n author = {Shepherd, N.E. and Gamsjaeger, R. and Vandevenne, M. and Cubeddu, L. and Mackay, J.P.},\\n doi = {10.1016/j.tet.2014.12.056},\\n journal = {Tetrahedron},\\n number = {5}\\n}\",\"author_short\":[\"Shepherd,  N.\",\"Gamsjaeger,  R.\",\"Vandevenne,  M.\",\"Cubeddu,  L.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"shepherd-gamsjaeger-vandevenne-cubeddu-mackay-sitedirectednitroxidespinlabelingofoligonucleotidesfornmrandeprstudies-2015\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells\",\"type\":\"article\",\"year\":\"2015\",\"pages\":\"960-965\",\"volume\":\"23\",\"id\":\"4e5561f0-05d0-3b6a-8a77-9e05d0b36591\",\"created\":\"2023-01-10T01:44:34.787Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:34.787Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).\",\"bibtype\":\"article\",\"author\":\"Saathoff, H. and Brofelth, M. and Trinh, A. and Parker, B.L. and Ryan, D.P. and Low, J.K.K. and Webb, S.R. and Silva, A.P.G. and Mackay, J.P. and Shepherd, N.E.\",\"doi\":\"10.1016/j.bmc.2015.01.023\",\"journal\":\"Bioorganic and Medicinal Chemistry\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells},\\n type = {article},\\n year = {2015},\\n pages = {960-965},\\n volume = {23},\\n id = {4e5561f0-05d0-3b6a-8a77-9e05d0b36591},\\n created = {2023-01-10T01:44:34.787Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:34.787Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).},\\n bibtype = {article},\\n author = {Saathoff, H. and Brofelth, M. and Trinh, A. and Parker, B.L. and Ryan, D.P. and Low, J.K.K. and Webb, S.R. and Silva, A.P.G. and Mackay, J.P. and Shepherd, N.E.},\\n doi = {10.1016/j.bmc.2015.01.023},\\n journal = {Bioorganic and Medicinal Chemistry},\\n number = {5}\\n}\",\"author_short\":[\"Saathoff,  H.\",\"Brofelth,  M.\",\"Trinh,  A.\",\"Parker,  B.\",\"Ryan,  D.\",\"Low,  J.\",\"Webb,  S.\",\"Silva,  A.\",\"Mackay,  J.\",\"Shepherd,  N.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The identification and structure of an N-terminal PR domain show that FOG1 is a member of the PRDM family of proteins\",\"type\":\"article\",\"year\":\"2014\",\"volume\":\"9\",\"id\":\"d4bd4375-43f0-3b26-8cf7-d8fffdd6c864\",\"created\":\"2023-01-10T01:44:35.718Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:35.718Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"FOG1 is a transcriptional regulator that acts in concert with the hematopoietic master regulator GATA1 to coordinate the differentiation of platelets and erythrocytes. Despite considerable effort, however, the mechanisms through which FOG1 regulates gene expression are only partially understood. Here we report the discovery of a previously unrecognized domain in FOG1: a PR (PRD-BF1 and RIZ) domain that is distantly related in sequence to the SET domains that are found in many histone methyltransferases. We have used NMR spectroscopy to determine the solution structure of this domain, revealing that the domain shares close structural similarity with SET domains. Titration with S-adenosyl-L-homocysteine, the cofactor product synonymous with SET domain methyltransferase activity, indicated that the FOG PR domain is not, however, likely to function as a methyltransferase in the same fashion. We also sought to define the function of this domain using both pulldown experiments and gel shift assays. However, neither pulldowns from mammalian nuclear extracts nor yeast two-hybrid assays reproducibly revealed binding partners, and we were unable to detect nucleic-acid-binding activity in this domain using our high-diversity Pentaprobe oligonucleotides. Overall, our data demonstrate that FOG1 is a member of the PRDM (PR domain containing proteins, with zinc fingers) family of transcriptional regulators. The function of many PR domains, however, remains somewhat enigmatic for the time being. © 2014 Clifton et al.\",\"bibtype\":\"article\",\"author\":\"Clifton, M.K. and Westman, B.J. and Thong, S.Y. and O'Connell, M.R. and Webster, M.W. and Shepherd, N.E. and Quinlan, K.G. and Crossley, M. and Blobel, G.A. and Mackay, J.P.\",\"doi\":\"10.1371/journal.pone.0106011\",\"journal\":\"PLoS ONE\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {The identification and structure of an N-terminal PR domain show that FOG1 is a member of the PRDM family of proteins},\\n type = {article},\\n year = {2014},\\n volume = {9},\\n id = {d4bd4375-43f0-3b26-8cf7-d8fffdd6c864},\\n created = {2023-01-10T01:44:35.718Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:35.718Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {FOG1 is a transcriptional regulator that acts in concert with the hematopoietic master regulator GATA1 to coordinate the differentiation of platelets and erythrocytes. Despite considerable effort, however, the mechanisms through which FOG1 regulates gene expression are only partially understood. Here we report the discovery of a previously unrecognized domain in FOG1: a PR (PRD-BF1 and RIZ) domain that is distantly related in sequence to the SET domains that are found in many histone methyltransferases. We have used NMR spectroscopy to determine the solution structure of this domain, revealing that the domain shares close structural similarity with SET domains. Titration with S-adenosyl-L-homocysteine, the cofactor product synonymous with SET domain methyltransferase activity, indicated that the FOG PR domain is not, however, likely to function as a methyltransferase in the same fashion. We also sought to define the function of this domain using both pulldown experiments and gel shift assays. However, neither pulldowns from mammalian nuclear extracts nor yeast two-hybrid assays reproducibly revealed binding partners, and we were unable to detect nucleic-acid-binding activity in this domain using our high-diversity Pentaprobe oligonucleotides. Overall, our data demonstrate that FOG1 is a member of the PRDM (PR domain containing proteins, with zinc fingers) family of transcriptional regulators. The function of many PR domains, however, remains somewhat enigmatic for the time being. © 2014 Clifton et al.},\\n bibtype = {article},\\n author = {Clifton, M.K. and Westman, B.J. and Thong, S.Y. and O'Connell, M.R. and Webster, M.W. and Shepherd, N.E. and Quinlan, K.G. and Crossley, M. and Blobel, G.A. and Mackay, J.P.},\\n doi = {10.1371/journal.pone.0106011},\\n journal = {PLoS ONE},\\n number = {8}\\n}\",\"author_short\":[\"Clifton,  M.\",\"Westman,  B.\",\"Thong,  S.\",\"O'Connell,  M.\",\"Webster,  M.\",\"Shepherd,  N.\",\"Quinlan,  K.\",\"Crossley,  M.\",\"Blobel,  G.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"clifton-westman-thong-oconnell-webster-shepherd-quinlan-crossley-etal-theidentificationandstructureofannterminalprdomainshowthatfog1isamemberoftheprdmfamilyofproteins-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Ca2+-induced PRE-NMR changes in the troponin complex reveal the possessive nature of the cardiac isoform for its regulatory switch\",\"type\":\"article\",\"year\":\"2014\",\"volume\":\"9\",\"id\":\"c3b7b413-fdec-325f-9bcd-b242670e45a7\",\"created\":\"2023-01-10T01:44:36.628Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:36.628Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The interaction between myosin and actin in cardiac muscle, modulated by the calcium (Ca2+) sensor Troponin complex (Tn), is a complex process which is yet to be fully resolved at the molecular level. Our understanding of how the binding of Ca2+ triggers conformational changes within Tn that are subsequently propagated through the contractile apparatus to initiate muscle activation is hampered by a lack of an atomic structure for the Ca2+-free state of the cardiac isoform. We have used paramagnetic relaxation enhancement (PRE)-NMR to obtain a description of the Ca2+-free state of cardiac Tn by describing the movement of key regions of the troponin I (cTnI) subunit upon the release of Ca2+ from Troponin C (cTnC). Site-directed spin-labeling was used to position paramagnetic spin labels in cTnI and the changes in the interaction between cTnI and cTnC subunits were then mapped by PRE-NMR. The functionally important regions of cTnI targeted in this study included the cTnC-binding N-region (cTnI57), the inhibitory region (cTnI143), and two sites on the regulatory switch region (cTnI151 and cTnI159). Comparison of 1H-15N-TROSY spectra of Ca2+-bound and free states for the spin labeled cTnCcTnI binary constructs demonstrated the release and modest movement of the cTnI switch region (,10 A ) away from the hydrophobic N-lobe of troponin C (cTnC) upon the removal of Ca2+. Our data supports a model where the non-bound regulatory switch region of cTnI is highly flexible in the absence of Ca2+ but remains in close vicinity to cTnC. We speculate that the close proximity of TnI to TnC in the cardiac complex is favourable for increasing the frequency of collisions between the N-lobe of cTnC and the regulatory switch region, counterbalancing the reduction in collision probability that results from the incomplete opening of the N-lobe of TnC that is unique to the cardiac isoform.\",\"bibtype\":\"article\",\"author\":\"Cordina, N.M. and Liew, C.K. and Potluri, P.R. and Curmi, P.M. and Fajer, P.G. and Logan, T.M. and Mackay, J.P. and Brown, L.J.\",\"doi\":\"10.1371/journal.pone.0112976\",\"journal\":\"PLoS ONE\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Ca2+-induced PRE-NMR changes in the troponin complex reveal the possessive nature of the cardiac isoform for its regulatory switch},\\n type = {article},\\n year = {2014},\\n volume = {9},\\n id = {c3b7b413-fdec-325f-9bcd-b242670e45a7},\\n created = {2023-01-10T01:44:36.628Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:36.628Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The interaction between myosin and actin in cardiac muscle, modulated by the calcium (Ca2+) sensor Troponin complex (Tn), is a complex process which is yet to be fully resolved at the molecular level. Our understanding of how the binding of Ca2+ triggers conformational changes within Tn that are subsequently propagated through the contractile apparatus to initiate muscle activation is hampered by a lack of an atomic structure for the Ca2+-free state of the cardiac isoform. We have used paramagnetic relaxation enhancement (PRE)-NMR to obtain a description of the Ca2+-free state of cardiac Tn by describing the movement of key regions of the troponin I (cTnI) subunit upon the release of Ca2+ from Troponin C (cTnC). Site-directed spin-labeling was used to position paramagnetic spin labels in cTnI and the changes in the interaction between cTnI and cTnC subunits were then mapped by PRE-NMR. The functionally important regions of cTnI targeted in this study included the cTnC-binding N-region (cTnI57), the inhibitory region (cTnI143), and two sites on the regulatory switch region (cTnI151 and cTnI159). Comparison of 1H-15N-TROSY spectra of Ca2+-bound and free states for the spin labeled cTnCcTnI binary constructs demonstrated the release and modest movement of the cTnI switch region (,10 A ) away from the hydrophobic N-lobe of troponin C (cTnC) upon the removal of Ca2+. Our data supports a model where the non-bound regulatory switch region of cTnI is highly flexible in the absence of Ca2+ but remains in close vicinity to cTnC. We speculate that the close proximity of TnI to TnC in the cardiac complex is favourable for increasing the frequency of collisions between the N-lobe of cTnC and the regulatory switch region, counterbalancing the reduction in collision probability that results from the incomplete opening of the N-lobe of TnC that is unique to the cardiac isoform.},\\n bibtype = {article},\\n author = {Cordina, N.M. and Liew, C.K. and Potluri, P.R. and Curmi, P.M. and Fajer, P.G. and Logan, T.M. and Mackay, J.P. and Brown, L.J.},\\n doi = {10.1371/journal.pone.0112976},\\n journal = {PLoS ONE},\\n number = {11}\\n}\",\"author_short\":[\"Cordina,  N.\",\"Liew,  C.\",\"Potluri,  P.\",\"Curmi,  P.\",\"Fajer,  P.\",\"Logan,  T.\",\"Mackay,  J.\",\"Brown,  L.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"cordina-liew-potluri-curmi-fajer-logan-mackay-brown-ca2inducedprenmrchangesinthetroponincomplexrevealthepossessivenatureofthecardiacisoformforitsregulatoryswitch-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Engineering specificity changes on a RanBP2 zinc finger that binds single-stranded RNA\",\"type\":\"article\",\"year\":\"2014\",\"pages\":\"7848-7852\",\"volume\":\"53\",\"id\":\"efff0e09-c7d0-32a8-9607-835d587fd71a\",\"created\":\"2023-01-10T01:44:37.599Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:37.599Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.\",\"bibtype\":\"article\",\"author\":\"Vandevenne, M. and O'Connell, M.R. and Helder, S. and Shepherd, N.E. and Matthews, J.M. and Kwan, A.H. and Segal, D.J. and Mackay, J.P.\",\"doi\":\"10.1002/anie.201402980\",\"journal\":\"Angewandte Chemie - International Edition\",\"number\":\"30\",\"bibtex\":\"@article{\\n title = {Engineering specificity changes on a RanBP2 zinc finger that binds single-stranded RNA},\\n type = {article},\\n year = {2014},\\n pages = {7848-7852},\\n volume = {53},\\n id = {efff0e09-c7d0-32a8-9607-835d587fd71a},\\n created = {2023-01-10T01:44:37.599Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:37.599Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.},\\n bibtype = {article},\\n author = {Vandevenne, M. and O'Connell, M.R. and Helder, S. and Shepherd, N.E. and Matthews, J.M. and Kwan, A.H. and Segal, D.J. and Mackay, J.P.},\\n doi = {10.1002/anie.201402980},\\n journal = {Angewandte Chemie - International Edition},\\n number = {30}\\n}\",\"author_short\":[\"Vandevenne,  M.\",\"O'Connell,  M.\",\"Helder,  S.\",\"Shepherd,  N.\",\"Matthews,  J.\",\"Kwan,  A.\",\"Segal,  D.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The structure of an LIM-Only protein 4 (LMO4) and Deformed Epidermal Autoregulatory Factor-1 (DEAF1) complex reveals a common mode of binding to LMO4\",\"type\":\"article\",\"year\":\"2014\",\"volume\":\"9\",\"id\":\"5973cfa4-e9ae-3e89-b66f-cf09132c9ab9\",\"created\":\"2023-01-10T01:44:38.507Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:38.507Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"LIM-domain only protein 4 (LMO4) is a widely expressed protein with important roles in embryonic development and breast cancer. It has been reported to bind many partners, including the transcription factor Deformed epidermal autoregulatory factor-1 (DEAF1), with which LMO4 shares many biological parallels. We used yeast two-hybrid assays to show that DEAF1 binds both LIM domains of LMO4 and that DEAF1 binds the same face on LMO4 as two other LMO4-binding partners, namely LIM domain binding protein 1 (LDB1) and C-terminal binding protein interacting protein (CtIP/RBBP8). Mutagenic screening analysed by the same method, indicates that the key residues in the interaction lie in LMO4LIM2and the N-terminal half of the LMO4-binding domain in DEAF1. We generated a stable LMO4LIM2-DEAF1 complex and determined the solution structure of that complex. Although the LMO4-binding domain from DEAF1 is intrinsically disordered, it becomes structured on binding. The structure confirms that LDB1, CtIP and DEAF1 all bind to the same face on LMO4. LMO4 appears to form a hub in protein-protein interaction networks, linking numerous pathways within cells. Competitive binding for LMO4 therefore most likely provides a level of regulation between those different pathways.\",\"bibtype\":\"article\",\"author\":\"Joseph, S. and Kwan, A.H. and Stokes, P.H. and Mackay, J.P. and Cubeddu, L. and Matthews, J.M.\",\"doi\":\"10.1371/journal.pone.0109108\",\"journal\":\"PLoS ONE\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {The structure of an LIM-Only protein 4 (LMO4) and Deformed Epidermal Autoregulatory Factor-1 (DEAF1) complex reveals a common mode of binding to LMO4},\\n type = {article},\\n year = {2014},\\n volume = {9},\\n id = {5973cfa4-e9ae-3e89-b66f-cf09132c9ab9},\\n created = {2023-01-10T01:44:38.507Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:38.507Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {LIM-domain only protein 4 (LMO4) is a widely expressed protein with important roles in embryonic development and breast cancer. It has been reported to bind many partners, including the transcription factor Deformed epidermal autoregulatory factor-1 (DEAF1), with which LMO4 shares many biological parallels. We used yeast two-hybrid assays to show that DEAF1 binds both LIM domains of LMO4 and that DEAF1 binds the same face on LMO4 as two other LMO4-binding partners, namely LIM domain binding protein 1 (LDB1) and C-terminal binding protein interacting protein (CtIP/RBBP8). Mutagenic screening analysed by the same method, indicates that the key residues in the interaction lie in LMO4LIM2and the N-terminal half of the LMO4-binding domain in DEAF1. We generated a stable LMO4LIM2-DEAF1 complex and determined the solution structure of that complex. Although the LMO4-binding domain from DEAF1 is intrinsically disordered, it becomes structured on binding. The structure confirms that LDB1, CtIP and DEAF1 all bind to the same face on LMO4. LMO4 appears to form a hub in protein-protein interaction networks, linking numerous pathways within cells. Competitive binding for LMO4 therefore most likely provides a level of regulation between those different pathways.},\\n bibtype = {article},\\n author = {Joseph, S. and Kwan, A.H. and Stokes, P.H. and Mackay, J.P. and Cubeddu, L. and Matthews, J.M.},\\n doi = {10.1371/journal.pone.0109108},\\n journal = {PLoS ONE},\\n number = {10}\\n}\",\"author_short\":[\"Joseph,  S.\",\"Kwan,  A.\",\"Stokes,  P.\",\"Mackay,  J.\",\"Cubeddu,  L.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"joseph-kwan-stokes-mackay-cubeddu-matthews-thestructureofanlimonlyprotein4lmo4anddeformedepidermalautoregulatoryfactor1deaf1complexrevealsacommonmodeofbindingtolmo4-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Insight into the architecture of the NuRD complex: Structure of the RbAp48-MTA1 subcomplex\",\"type\":\"article\",\"year\":\"2014\",\"pages\":\"21844-21855\",\"volume\":\"289\",\"id\":\"a0afaf4d-99c7-3fea-9a0a-a55fcb967c4a\",\"created\":\"2023-01-10T01:44:39.545Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:39.545Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background: The NuRD complex controls gene expression through altering chromatin structure. Results: The MTA1-RbAp48 structure shows how the RbAp46/p48 histone chaperones are recruited to NuRD. Conclusion: The MTA subunits act as scaffolds for NuRD complex assembly. Significance: The MTA/RbAp48 interaction prevents binding of histone H4, which is crucial for understanding the role of the RbAp46/p48 chaperones in the complex. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Alqarni, S.S.M. and Murthy, A. and Zhang, W. and Przewloka, M.R. and Silva, A.P.G. and Watson, A.A. and Lejon, S. and Pei, X.Y. and Smits, A.H. and Kloet, S.L. and Mackay, J.P. and Laue, E.D.\",\"doi\":\"10.1074/jbc.M114.558940\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"32\",\"bibtex\":\"@article{\\n title = {Insight into the architecture of the NuRD complex: Structure of the RbAp48-MTA1 subcomplex},\\n type = {article},\\n year = {2014},\\n pages = {21844-21855},\\n volume = {289},\\n id = {a0afaf4d-99c7-3fea-9a0a-a55fcb967c4a},\\n created = {2023-01-10T01:44:39.545Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:39.545Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background: The NuRD complex controls gene expression through altering chromatin structure. Results: The MTA1-RbAp48 structure shows how the RbAp46/p48 histone chaperones are recruited to NuRD. Conclusion: The MTA subunits act as scaffolds for NuRD complex assembly. Significance: The MTA/RbAp48 interaction prevents binding of histone H4, which is crucial for understanding the role of the RbAp46/p48 chaperones in the complex. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Alqarni, S.S.M. and Murthy, A. and Zhang, W. and Przewloka, M.R. and Silva, A.P.G. and Watson, A.A. and Lejon, S. and Pei, X.Y. and Smits, A.H. and Kloet, S.L. and Mackay, J.P. and Laue, E.D.},\\n doi = {10.1074/jbc.M114.558940},\\n journal = {Journal of Biological Chemistry},\\n number = {32}\\n}\",\"author_short\":[\"Alqarni,  S.\",\"Murthy,  A.\",\"Zhang,  W.\",\"Przewloka,  M.\",\"Silva,  A.\",\"Watson,  A.\",\"Lejon,  S.\",\"Pei,  X.\",\"Smits,  A.\",\"Kloet,  S.\",\"Mackay,  J.\",\"Laue,  E.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"alqarni-murthy-zhang-przewloka-silva-watson-lejon-pei-etal-insightintothearchitectureofthenurdcomplexstructureoftherbap48mta1subcomplex-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Structure of the hemoglobin-isdh complex reveals the molecular basis of iron capture by staphylococcus aureus\",\"type\":\"article\",\"year\":\"2014\",\"pages\":\"6728-6738\",\"volume\":\"289\",\"id\":\"8082b276-9693-3b97-b289-93ddb85c7c38\",\"created\":\"2023-01-10T01:44:40.488Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:40.488Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background: IsdB and IsdH proteins from Staphylococcus aureus strip heme iron from human hemoglobin. Results: The IsdH·hemoglobin complex shows how globin-binding and heme-binding NEAT domains of IsdH cooperate to remove heme from both chains of hemoglobin. Conclusion: The supradomain architecture of IsdH confers activity by precisely positioning the heme acceptor domain. Significance: Multiple IsdH·hemoglobin interfaces may be targets for new antibiotics. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Dickson, C.F. and Kumar, K.K. and Jacques, D.A. and Malmirchegini, G.R. and Spirig, T. and Mackay, J.P. and Clubb, R.T. and Guss, J.M. and Gell, D.A.\",\"doi\":\"10.1074/jbc.M113.545566\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {Structure of the hemoglobin-isdh complex reveals the molecular basis of iron capture by staphylococcus aureus},\\n type = {article},\\n year = {2014},\\n pages = {6728-6738},\\n volume = {289},\\n id = {8082b276-9693-3b97-b289-93ddb85c7c38},\\n created = {2023-01-10T01:44:40.488Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:40.488Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background: IsdB and IsdH proteins from Staphylococcus aureus strip heme iron from human hemoglobin. Results: The IsdH·hemoglobin complex shows how globin-binding and heme-binding NEAT domains of IsdH cooperate to remove heme from both chains of hemoglobin. Conclusion: The supradomain architecture of IsdH confers activity by precisely positioning the heme acceptor domain. Significance: Multiple IsdH·hemoglobin interfaces may be targets for new antibiotics. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Dickson, C.F. and Kumar, K.K. and Jacques, D.A. and Malmirchegini, G.R. and Spirig, T. and Mackay, J.P. and Clubb, R.T. and Guss, J.M. and Gell, D.A.},\\n doi = {10.1074/jbc.M113.545566},\\n journal = {Journal of Biological Chemistry},\\n number = {10}\\n}\",\"author_short\":[\"Dickson,  C.\",\"Kumar,  K.\",\"Jacques,  D.\",\"Malmirchegini,  G.\",\"Spirig,  T.\",\"Mackay,  J.\",\"Clubb,  R.\",\"Guss,  J.\",\"Gell,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dickson-kumar-jacques-malmirchegini-spirig-mackay-clubb-guss-etal-structureofthehemoglobinisdhcomplexrevealsthemolecularbasisofironcapturebystaphylococcusaureus-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5\",\"type\":\"article\",\"year\":\"2014\",\"pages\":\"54-60\",\"volume\":\"100\",\"id\":\"79bfac53-430c-3f33-98e9-37db63509339\",\"created\":\"2023-01-10T01:44:41.424Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:41.424Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Blythe, A. and Gunasekara, S. and Walshe, J. and Mackay, J.P. and Hartzog, G.A. and Vrielink, A.\",\"doi\":\"10.1016/j.pep.2014.05.005\",\"journal\":\"Protein Expression and Purification\",\"bibtex\":\"@article{\\n title = {Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5},\\n type = {article},\\n year = {2014},\\n pages = {54-60},\\n volume = {100},\\n id = {79bfac53-430c-3f33-98e9-37db63509339},\\n created = {2023-01-10T01:44:41.424Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:41.424Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.},\\n bibtype = {article},\\n author = {Blythe, A. and Gunasekara, S. and Walshe, J. and Mackay, J.P. and Hartzog, G.A. and Vrielink, A.},\\n doi = {10.1016/j.pep.2014.05.005},\\n journal = {Protein Expression and Purification}\\n}\",\"author_short\":[\"Blythe,  A.\",\"Gunasekara,  S.\",\"Walshe,  J.\",\"Mackay,  J.\",\"Hartzog,  G.\",\"Vrielink,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1\",\"type\":\"article\",\"year\":\"2014\",\"pages\":\"141-144\",\"volume\":\"8\",\"id\":\"a64344fa-3b32-37f0-b1e1-f7b7ad8cbfd8\",\"created\":\"2023-01-10T01:44:42.330Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:42.330Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.\",\"bibtype\":\"article\",\"author\":\"Joseph, S. and Kwan, A.H.Y. and Mackay, J.P. and Cubeddu, L. and Matthews, J.M.\",\"doi\":\"10.1007/s12104-013-9470-x\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1},\\n type = {article},\\n year = {2014},\\n pages = {141-144},\\n volume = {8},\\n id = {a64344fa-3b32-37f0-b1e1-f7b7ad8cbfd8},\\n created = {2023-01-10T01:44:42.330Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:42.330Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.},\\n bibtype = {article},\\n author = {Joseph, S. and Kwan, A.H.Y. and Mackay, J.P. and Cubeddu, L. and Matthews, J.M.},\\n doi = {10.1007/s12104-013-9470-x},\\n journal = {Biomolecular NMR Assignments},\\n number = {1}\\n}\",\"author_short\":[\"Joseph,  S.\",\"Kwan,  A.\",\"Mackay,  J.\",\"Cubeddu,  L.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Trim58 Degrades Dynein and Regulates Terminal Erythropoiesis\",\"type\":\"article\",\"year\":\"2014\",\"pages\":\"688-700\",\"volume\":\"30\",\"id\":\"c1ccb790-7e19-39e4-aaad-2c8dd2dc11ef\",\"created\":\"2023-01-10T01:44:43.258Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:43.258Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"TRIM58 is an E3 ubiquitin ligase superfamily member implicated by genome-wide association studies to regulate human erythrocyte traits. Here, we show that Trim58 expression is induced during late erythropoiesis and that its depletion by small hairpin RNAs (shRNAs) inhibits the maturation of late-stage nucleated erythroblasts to anucleate reticulocytes. Imaging flow cytometry studies demonstrate that Trim58 regulates polarization and/or extrusion of erythroblast nuclei. Invitro, Trim58 directly binds and ubiquitinates the intermediate chain of the microtubule motor dynein. In cells, Trim58 stimulates proteasome-dependent degradation of the dynein holoprotein complex. During erythropoiesis, Trim58 expression, dynein loss, and enucleation occur concomitantly, and all are inhibited by Trim58 shRNAs. Dynein regulates nuclear positioning and microtubule organization, both of which undergo dramatic changes during erythroblast enucleation. Thus, we propose that Trim58 promotes this processby eliminating dynein. Our findings identify an erythroid-specific regulator of enucleation and elucidate a previously unrecognized mechanism for controlling dynein activity.\",\"bibtype\":\"article\",\"author\":\"Thom, C.S. and Traxler, E.A. and Khandros, E. and Nickas, J.M. and Zhou, O.Y. and Lazarus, J.E. and Silva, A.P.G. and Prabhu, D. and Yao, Y. and Aribeana, C. and Holzbaur, E.L.F. and Weiss, M.J.\",\"doi\":\"10.1016/j.devcel.2014.07.021\",\"journal\":\"Developmental Cell\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Trim58 Degrades Dynein and Regulates Terminal Erythropoiesis},\\n type = {article},\\n year = {2014},\\n pages = {688-700},\\n volume = {30},\\n id = {c1ccb790-7e19-39e4-aaad-2c8dd2dc11ef},\\n created = {2023-01-10T01:44:43.258Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:43.258Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {TRIM58 is an E3 ubiquitin ligase superfamily member implicated by genome-wide association studies to regulate human erythrocyte traits. Here, we show that Trim58 expression is induced during late erythropoiesis and that its depletion by small hairpin RNAs (shRNAs) inhibits the maturation of late-stage nucleated erythroblasts to anucleate reticulocytes. Imaging flow cytometry studies demonstrate that Trim58 regulates polarization and/or extrusion of erythroblast nuclei. Invitro, Trim58 directly binds and ubiquitinates the intermediate chain of the microtubule motor dynein. In cells, Trim58 stimulates proteasome-dependent degradation of the dynein holoprotein complex. During erythropoiesis, Trim58 expression, dynein loss, and enucleation occur concomitantly, and all are inhibited by Trim58 shRNAs. Dynein regulates nuclear positioning and microtubule organization, both of which undergo dramatic changes during erythroblast enucleation. Thus, we propose that Trim58 promotes this processby eliminating dynein. Our findings identify an erythroid-specific regulator of enucleation and elucidate a previously unrecognized mechanism for controlling dynein activity.},\\n bibtype = {article},\\n author = {Thom, C.S. and Traxler, E.A. and Khandros, E. and Nickas, J.M. and Zhou, O.Y. and Lazarus, J.E. and Silva, A.P.G. and Prabhu, D. and Yao, Y. and Aribeana, C. and Holzbaur, E.L.F. and Weiss, M.J.},\\n doi = {10.1016/j.devcel.2014.07.021},\\n journal = {Developmental Cell},\\n number = {6}\\n}\",\"author_short\":[\"Thom,  C.\",\"Traxler,  E.\",\"Khandros,  E.\",\"Nickas,  J.\",\"Zhou,  O.\",\"Lazarus,  J.\",\"Silva,  A.\",\"Prabhu,  D.\",\"Yao,  Y.\",\"Aribeana,  C.\",\"Holzbaur,  E.\",\"Weiss,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"thom-traxler-khandros-nickas-zhou-lazarus-silva-prabhu-etal-trim58degradesdyneinandregulatesterminalerythropoiesis-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Transcription factor seeks DNA - Cognate site preferred\",\"type\":\"article\",\"year\":\"2014\",\"pages\":\"1370-1372\",\"volume\":\"426\",\"id\":\"1e075f30-6417-39c7-8b24-b13d5c28f504\",\"created\":\"2023-01-10T01:44:44.178Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:44.178Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Mackay, J.\",\"doi\":\"10.1016/j.jmb.2013.12.010\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {Transcription factor seeks DNA - Cognate site preferred},\\n type = {article},\\n year = {2014},\\n pages = {1370-1372},\\n volume = {426},\\n id = {1e075f30-6417-39c7-8b24-b13d5c28f504},\\n created = {2023-01-10T01:44:44.178Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:44.178Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Mackay, J.},\\n doi = {10.1016/j.jmb.2013.12.010},\\n journal = {Journal of Molecular Biology},\\n number = {7}\\n}\",\"author_short\":[\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-transcriptionfactorseeksdnacognatesitepreferred-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of an N-terminal domain of CHD4\",\"type\":\"article\",\"year\":\"2014\",\"pages\":\"137-139\",\"volume\":\"8\",\"id\":\"cd0244ce-1944-3196-a041-f999ed20f6c3\",\"created\":\"2023-01-10T01:44:45.086Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:45.086Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.\",\"bibtype\":\"article\",\"author\":\"Silva, A.P.G. and Kwan, A.H. and Mackay, J.P.\",\"doi\":\"10.1007/s12104-013-9469-3\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of an N-terminal domain of CHD4},\\n type = {article},\\n year = {2014},\\n pages = {137-139},\\n volume = {8},\\n id = {cd0244ce-1944-3196-a041-f999ed20f6c3},\\n created = {2023-01-10T01:44:45.086Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:45.086Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.},\\n bibtype = {article},\\n author = {Silva, A.P.G. and Kwan, A.H. and Mackay, J.P.},\\n doi = {10.1007/s12104-013-9469-3},\\n journal = {Biomolecular NMR Assignments},\\n number = {1}\\n}\",\"author_short\":[\"Silva,  A.\",\"Kwan,  A.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"silva-kwan-mackay-sup1suphsup13supcandsup15supnresonanceassignmentsofannterminaldomainofchd4-2014\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"α-Hemoglobin-stabilizing Protein (AHSP) perturbs the proximal heme pocket of oxy-α-hemoglobin and weakens the iron-oxygen bond\",\"type\":\"article\",\"year\":\"2013\",\"pages\":\"19986-20001\",\"volume\":\"288\",\"id\":\"d5cb8d14-4ed7-37da-a53b-1987811758ed\",\"created\":\"2023-01-10T01:44:46.024Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:46.024Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"α-Hemoglobin (αHb)-stabilizing protein (AHSP) is a molecular chaperone that assists hemoglobin assembly. AHSP induces changes in αHb heme coordination, but how these changes are facilitated by interactions at the αHb·AHSP interface is not well understood. To address this question we have used NMR, x-ray absorption spectroscopy, and ligand binding measurements to probe αHb conformational changes induced by AHSP binding.NMRchemical shift analyses of free CO-αHb and CO- αHb·AHSP indicated that the seven helical elements of the native αHb structure are retained and that the heme Fe(II) remains coordinated to the proximal His-87 side chain. However, chemical shift differences revealed alterations of the F, G, and H helices and the heme pocket of CO-αHb bound to AHSP. Comparisons of iron-ligand geometry using extended x-ray absorption fine structure spectroscopy showed that AHSP binding induces a small 0.03 Å lengthening of the Fe-O2 bond, explaining previous reports that AHSP decreases αHb O2 affinity roughly 4-fold and promotes autooxidation due primarily to a 3-4-fold increase in the rate ofO2 dissociation. Pro-30 mutations diminished NMR chemical shift changes in the proximal heme pocket, restored normal O2 dissociation rate and equilibrium constants, and reduced O2-αHb autooxidation rates. Thus, the contacts mediated by Pro-30 in wild-type AHSP promote αHb autooxidation by introducing strain into the proximal heme pocket. As a chaperone, AHSP facilitates rapid assembly of αHb into Hb when βHb is abundant but diverts αHb to a redox resistant holding state when βHb is limiting. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Dickson, C.F. and Rich, A.M. and D'Avigdor, W.M.H. and Collins, D.A.T. and Lowry, J.A. and Mollan, T.L. and Khandros, E. and Olson, J.S. and Weiss, M.J. and MacKay, J.P. and Lay, P.A. and Gell, D.A.\",\"doi\":\"10.1074/jbc.M112.437509\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"27\",\"bibtex\":\"@article{\\n title = {α-Hemoglobin-stabilizing Protein (AHSP) perturbs the proximal heme pocket of oxy-α-hemoglobin and weakens the iron-oxygen bond},\\n type = {article},\\n year = {2013},\\n pages = {19986-20001},\\n volume = {288},\\n id = {d5cb8d14-4ed7-37da-a53b-1987811758ed},\\n created = {2023-01-10T01:44:46.024Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:46.024Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {α-Hemoglobin (αHb)-stabilizing protein (AHSP) is a molecular chaperone that assists hemoglobin assembly. AHSP induces changes in αHb heme coordination, but how these changes are facilitated by interactions at the αHb·AHSP interface is not well understood. To address this question we have used NMR, x-ray absorption spectroscopy, and ligand binding measurements to probe αHb conformational changes induced by AHSP binding.NMRchemical shift analyses of free CO-αHb and CO- αHb·AHSP indicated that the seven helical elements of the native αHb structure are retained and that the heme Fe(II) remains coordinated to the proximal His-87 side chain. However, chemical shift differences revealed alterations of the F, G, and H helices and the heme pocket of CO-αHb bound to AHSP. Comparisons of iron-ligand geometry using extended x-ray absorption fine structure spectroscopy showed that AHSP binding induces a small 0.03 Å lengthening of the Fe-O2 bond, explaining previous reports that AHSP decreases αHb O2 affinity roughly 4-fold and promotes autooxidation due primarily to a 3-4-fold increase in the rate ofO2 dissociation. Pro-30 mutations diminished NMR chemical shift changes in the proximal heme pocket, restored normal O2 dissociation rate and equilibrium constants, and reduced O2-αHb autooxidation rates. Thus, the contacts mediated by Pro-30 in wild-type AHSP promote αHb autooxidation by introducing strain into the proximal heme pocket. As a chaperone, AHSP facilitates rapid assembly of αHb into Hb when βHb is abundant but diverts αHb to a redox resistant holding state when βHb is limiting. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Dickson, C.F. and Rich, A.M. and D'Avigdor, W.M.H. and Collins, D.A.T. and Lowry, J.A. and Mollan, T.L. and Khandros, E. and Olson, J.S. and Weiss, M.J. and MacKay, J.P. and Lay, P.A. and Gell, D.A.},\\n doi = {10.1074/jbc.M112.437509},\\n journal = {Journal of Biological Chemistry},\\n number = {27}\\n}\",\"author_short\":[\"Dickson,  C.\",\"Rich,  A.\",\"D'Avigdor,  W.\",\"Collins,  D.\",\"Lowry,  J.\",\"Mollan,  T.\",\"Khandros,  E.\",\"Olson,  J.\",\"Weiss,  M.\",\"MacKay,  J.\",\"Lay,  P.\",\"Gell,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dickson-rich-davigdor-collins-lowry-mollan-khandros-olson-etal-hemoglobinstabilizingproteinahspperturbstheproximalhemepocketofoxyhemoglobinandweakenstheironoxygenbond-2013\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Is there a telltale RH fingerprint in zinc fingers that recognizes methylated CpG dinucleotides?\",\"type\":\"article\",\"year\":\"2013\",\"pages\":\"421-422\",\"volume\":\"38\",\"id\":\"4f05c721-c533-3f78-93e4-e5649cccf00b\",\"created\":\"2023-01-10T01:44:46.983Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:46.983Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Segal, D.J. and Crossley, M.\",\"doi\":\"10.1016/j.tibs.2013.06.005\",\"journal\":\"Trends in Biochemical Sciences\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {Is there a telltale RH fingerprint in zinc fingers that recognizes methylated CpG dinucleotides?},\\n type = {article},\\n year = {2013},\\n pages = {421-422},\\n volume = {38},\\n id = {4f05c721-c533-3f78-93e4-e5649cccf00b},\\n created = {2023-01-10T01:44:46.983Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:46.983Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Mackay, J.P. and Segal, D.J. and Crossley, M.},\\n doi = {10.1016/j.tibs.2013.06.005},\\n journal = {Trends in Biochemical Sciences},\\n number = {9}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Segal,  D.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-segal-crossley-isthereatelltalerhfingerprintinzincfingersthatrecognizesmethylatedcpgdinucleotides-2013\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A structural analysis of DNA binding by myelin transcription factor 1 double zinc fingers\",\"type\":\"article\",\"year\":\"2013\",\"pages\":\"35180-35191\",\"volume\":\"288\",\"id\":\"87086ec5-3591-3e63-826a-b7a3fde32906\",\"created\":\"2023-01-10T01:44:47.925Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:47.925Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background: Myelin transcription factor 1 (MyT1) contains seven similar zinc finger domains that bind DNA specifically. Results: A three-dimensional structural model explains how a double zinc finger unit is able to recognize DNA. Conclusion: DNA-binding residues are conserved among all MyT1 zinc fingers, suggesting an identical DNA binding mode. Significance: Determination of the molecular details of DNA interaction will be crucial in understanding MyT1 function. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Gamsjaeger, R. and O'Connell, M.R. and Cubeddu, L. and Shepherd, N.E. and Lowry, J.A. and Kwan, A.H. and Vandevenne, M. and Swanton, M.K. and Matthews, J.M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M113.482075\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"49\",\"bibtex\":\"@article{\\n title = {A structural analysis of DNA binding by myelin transcription factor 1 double zinc fingers},\\n type = {article},\\n year = {2013},\\n pages = {35180-35191},\\n volume = {288},\\n id = {87086ec5-3591-3e63-826a-b7a3fde32906},\\n created = {2023-01-10T01:44:47.925Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:47.925Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background: Myelin transcription factor 1 (MyT1) contains seven similar zinc finger domains that bind DNA specifically. Results: A three-dimensional structural model explains how a double zinc finger unit is able to recognize DNA. Conclusion: DNA-binding residues are conserved among all MyT1 zinc fingers, suggesting an identical DNA binding mode. Significance: Determination of the molecular details of DNA interaction will be crucial in understanding MyT1 function. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Gamsjaeger, R. and O'Connell, M.R. and Cubeddu, L. and Shepherd, N.E. and Lowry, J.A. and Kwan, A.H. and Vandevenne, M. and Swanton, M.K. and Matthews, J.M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M113.482075},\\n journal = {Journal of Biological Chemistry},\\n number = {49}\\n}\",\"author_short\":[\"Gamsjaeger,  R.\",\"O'Connell,  M.\",\"Cubeddu,  L.\",\"Shepherd,  N.\",\"Lowry,  J.\",\"Kwan,  A.\",\"Vandevenne,  M.\",\"Swanton,  M.\",\"Matthews,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gamsjaeger-oconnell-cubeddu-shepherd-lowry-kwan-vandevenne-swanton-etal-astructuralanalysisofdnabindingbymyelintranscriptionfactor1doublezincfingers-2013\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"New insights into DNA recognition by zinc fingers revealed by structural analysis of the oncoprotein ZNF217\",\"type\":\"article\",\"year\":\"2013\",\"pages\":\"10616-10627\",\"volume\":\"288\",\"id\":\"406b142f-54cb-305c-8eaa-3b8866da36f3\",\"created\":\"2023-01-10T01:44:48.847Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:48.847Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background: Classical zinc finger proteins are extremely abundant and interact with DNA using a well defined recognition code. Results: We solved the structure of ZNF217 bound to its cognate DNA. Conclusion: ZNF217 presents a unique DNA interaction pattern including a new type of protein-DNA contact. Significance: This study deepens our understanding of DNA recognition by classical zinc fingers. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Vandevenne, M. and Jacques, D.A. and Artuz, C. and Nguyen, C.D. and Kwan, A.H.Y. and Segal, D.J. and Matthews, J.M. and Crossley, M. and Guss, J.M. and MacKay, J.P.\",\"doi\":\"10.1074/jbc.M112.441451\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"15\",\"bibtex\":\"@article{\\n title = {New insights into DNA recognition by zinc fingers revealed by structural analysis of the oncoprotein ZNF217},\\n type = {article},\\n year = {2013},\\n pages = {10616-10627},\\n volume = {288},\\n id = {406b142f-54cb-305c-8eaa-3b8866da36f3},\\n created = {2023-01-10T01:44:48.847Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:48.847Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background: Classical zinc finger proteins are extremely abundant and interact with DNA using a well defined recognition code. Results: We solved the structure of ZNF217 bound to its cognate DNA. Conclusion: ZNF217 presents a unique DNA interaction pattern including a new type of protein-DNA contact. Significance: This study deepens our understanding of DNA recognition by classical zinc fingers. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Vandevenne, M. and Jacques, D.A. and Artuz, C. and Nguyen, C.D. and Kwan, A.H.Y. and Segal, D.J. and Matthews, J.M. and Crossley, M. and Guss, J.M. and MacKay, J.P.},\\n doi = {10.1074/jbc.M112.441451},\\n journal = {Journal of Biological Chemistry},\\n number = {15}\\n}\",\"author_short\":[\"Vandevenne,  M.\",\"Jacques,  D.\",\"Artuz,  C.\",\"Nguyen,  C.\",\"Kwan,  A.\",\"Segal,  D.\",\"Matthews,  J.\",\"Crossley,  M.\",\"Guss,  J.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"vandevenne-jacques-artuz-nguyen-kwan-segal-matthews-crossley-etal-newinsightsintodnarecognitionbyzincfingersrevealedbystructuralanalysisoftheoncoproteinznf217-2013\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Effects of calcium binding and the hypertrophic cardiomyopathy A8V mutation on the dynamic equilibrium between closed and open conformations of the regulatory N-domain of isolated cardiac troponin C\",\"type\":\"article\",\"year\":\"2013\",\"pages\":\"1950-1962\",\"volume\":\"52\",\"id\":\"a6e23941-429e-3ffe-905a-69c9fa9461ee\",\"created\":\"2023-01-10T01:44:49.760Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:49.760Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Troponin C (TnC) is the calcium-binding subunit of the troponin complex responsible for initiating striated muscle contraction in response to calcium influx. In the skeletal TnC isoform, calcium binding induces a structural change in the regulatory N-domain of TnC that involves a transition from a closed to open structural state and accompanying exposure of a large hydrophobic patch for troponin I (TnI) to subsequently bind. However, little is understood about how calcium primes the N-domain of the cardiac isoform (cTnC) for interaction with the TnI subunit as the open conformation of the regulatory domain of cTnC has been observed only in the presence of bound TnI. Here we use paramagnetic relaxation enhancement (PRE) to characterize the closed to open transition of isolated cTnC in solution, a process that cannot be observed by traditional nuclear magnetic resonance methods. Our PRE data from four spin-labeled monocysteine constructs of isolated cTnC reveal that calcium binding triggers movement of the N-domain helices toward an open state. Fitting of the PRE data to a closed to open transition model reveals the presence of a small population of cTnC molecules in the absence of calcium that possess an open conformation, the level of which increases substantially upon Ca2+ binding. These data support a model in which calcium binding creates a dynamic equilibrium between the closed and open structural states to prime cTnC for interaction with its target peptide. We also used PRE data to assess the structural effects of a familial hypertrophic cardiomyopathy point mutation located within the N-domain of cTnC (A8V). The PRE data show that the Ca2+ switch mechanism is perturbed by the A8V mutation, resulting in a more open N-domain conformation in both the apo and holo states. © 2013 American Chemical Society.\",\"bibtype\":\"article\",\"author\":\"Cordina, N.M. and Liew, C.K. and Gell, D.A. and Fajer, P.G. and MacKay, J.P. and Brown, L.J.\",\"doi\":\"10.1021/bi4000172\",\"journal\":\"Biochemistry\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Effects of calcium binding and the hypertrophic cardiomyopathy A8V mutation on the dynamic equilibrium between closed and open conformations of the regulatory N-domain of isolated cardiac troponin C},\\n type = {article},\\n year = {2013},\\n pages = {1950-1962},\\n volume = {52},\\n id = {a6e23941-429e-3ffe-905a-69c9fa9461ee},\\n created = {2023-01-10T01:44:49.760Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:49.760Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Troponin C (TnC) is the calcium-binding subunit of the troponin complex responsible for initiating striated muscle contraction in response to calcium influx. In the skeletal TnC isoform, calcium binding induces a structural change in the regulatory N-domain of TnC that involves a transition from a closed to open structural state and accompanying exposure of a large hydrophobic patch for troponin I (TnI) to subsequently bind. However, little is understood about how calcium primes the N-domain of the cardiac isoform (cTnC) for interaction with the TnI subunit as the open conformation of the regulatory domain of cTnC has been observed only in the presence of bound TnI. Here we use paramagnetic relaxation enhancement (PRE) to characterize the closed to open transition of isolated cTnC in solution, a process that cannot be observed by traditional nuclear magnetic resonance methods. Our PRE data from four spin-labeled monocysteine constructs of isolated cTnC reveal that calcium binding triggers movement of the N-domain helices toward an open state. Fitting of the PRE data to a closed to open transition model reveals the presence of a small population of cTnC molecules in the absence of calcium that possess an open conformation, the level of which increases substantially upon Ca2+ binding. These data support a model in which calcium binding creates a dynamic equilibrium between the closed and open structural states to prime cTnC for interaction with its target peptide. We also used PRE data to assess the structural effects of a familial hypertrophic cardiomyopathy point mutation located within the N-domain of cTnC (A8V). The PRE data show that the Ca2+ switch mechanism is perturbed by the A8V mutation, resulting in a more open N-domain conformation in both the apo and holo states. © 2013 American Chemical Society.},\\n bibtype = {article},\\n author = {Cordina, N.M. and Liew, C.K. and Gell, D.A. and Fajer, P.G. and MacKay, J.P. and Brown, L.J.},\\n doi = {10.1021/bi4000172},\\n journal = {Biochemistry},\\n number = {11}\\n}\",\"author_short\":[\"Cordina,  N.\",\"Liew,  C.\",\"Gell,  D.\",\"Fajer,  P.\",\"MacKay,  J.\",\"Brown,  L.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"cordina-liew-gell-fajer-mackay-brown-effectsofcalciumbindingandthehypertrophiccardiomyopathya8vmutationonthedynamicequilibriumbetweenclosedandopenconformationsoftheregulatoryndomainofisolatedcardiactroponinc-2013\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural basis of the interaction of the breast cancer Oncogene LMO4 with the tumour suppressor CtIP/RBBP8\",\"type\":\"article\",\"year\":\"2013\",\"pages\":\"1101-1110\",\"volume\":\"425\",\"id\":\"191b6ccd-b3f5-3f12-a76e-f0e33183952f\",\"created\":\"2023-01-10T01:44:50.675Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:50.675Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"LIM-only protein 4 (LMO4) is strongly linked to the progression of breast cancer. Although the mechanisms underlying this phenomenon are not well understood, a role is emerging for LMO4 in regulation of the cell cycle. We determined the solution structure of LMO4 in complex with CtIP (C-terminal binding protein interacting protein)/RBBP8, a tumour suppressor protein that is involved in cell cycle progression, DNA repair and transcriptional regulation. Our data reveal that CtIP and the essential LMO cofactor LDB1 (LIM-domain binding protein 1) bind to the same face on LMO4 and cannot simultaneously bind to LMO4. We hypothesise that overexpression of LMO4 may disrupt some of the normal tumour suppressor activities of CtIP, thereby contributing to breast cancer progression. © 2013 Elsevier Ltd.\",\"bibtype\":\"article\",\"author\":\"Stokes, P.H. and Liew, C.W. and Kwan, A.H. and Foo, P. and Barker, H.E. and Djamirze, A. and O'Reilly, V. and Visvader, J.E. and Mackay, J.P. and Matthews, J.M.\",\"doi\":\"10.1016/j.jmb.2013.01.017\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {Structural basis of the interaction of the breast cancer Oncogene LMO4 with the tumour suppressor CtIP/RBBP8},\\n type = {article},\\n year = {2013},\\n pages = {1101-1110},\\n volume = {425},\\n id = {191b6ccd-b3f5-3f12-a76e-f0e33183952f},\\n created = {2023-01-10T01:44:50.675Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:50.675Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {LIM-only protein 4 (LMO4) is strongly linked to the progression of breast cancer. Although the mechanisms underlying this phenomenon are not well understood, a role is emerging for LMO4 in regulation of the cell cycle. We determined the solution structure of LMO4 in complex with CtIP (C-terminal binding protein interacting protein)/RBBP8, a tumour suppressor protein that is involved in cell cycle progression, DNA repair and transcriptional regulation. Our data reveal that CtIP and the essential LMO cofactor LDB1 (LIM-domain binding protein 1) bind to the same face on LMO4 and cannot simultaneously bind to LMO4. We hypothesise that overexpression of LMO4 may disrupt some of the normal tumour suppressor activities of CtIP, thereby contributing to breast cancer progression. © 2013 Elsevier Ltd.},\\n bibtype = {article},\\n author = {Stokes, P.H. and Liew, C.W. and Kwan, A.H. and Foo, P. and Barker, H.E. and Djamirze, A. and O'Reilly, V. and Visvader, J.E. and Mackay, J.P. and Matthews, J.M.},\\n doi = {10.1016/j.jmb.2013.01.017},\\n journal = {Journal of Molecular Biology},\\n number = {7}\\n}\",\"author_short\":[\"Stokes,  P.\",\"Liew,  C.\",\"Kwan,  A.\",\"Foo,  P.\",\"Barker,  H.\",\"Djamirze,  A.\",\"O'Reilly,  V.\",\"Visvader,  J.\",\"Mackay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"stokes-liew-kwan-foo-barker-djamirze-oreilly-visvader-etal-structuralbasisoftheinteractionofthebreastcanceroncogenelmo4withthetumoursuppressorctiprbbp8-2013\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Semiquantitative and quantitative analysis of protein-DNA interactions using steady-state measurements in surface plasmon resonance competition experiments\",\"type\":\"article\",\"year\":\"2013\",\"pages\":\"178-185\",\"volume\":\"440\",\"id\":\"cb78304b-2ce1-32bf-ae2e-25e278ca3b2c\",\"created\":\"2023-01-10T01:44:51.607Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:51.607Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"One method commonly used to characterize protein-DNA interactions is surface plasmon resonance (SPR). In a typical SPR experiment, chip-bound DNA is exposed to increasing concentrations of protein; the resulting binding data are used to calculate a dissociation constant for the interaction. However, in cases in which knowledge of the specificity of the interaction is required, a large set of DNA variants has to be tested; this is time consuming and costly, in part because of the requirement for multiple SPR chips. We have developed a new protocol that uses steady-state binding levels in SPR competition experiments to determine protein-binding dissociation constants for a set of DNA variants. This approach is rapid and straightforward and requires the use of only a single SPR chip. Additionally, in contrast to other methods, our approach does not require prior knowledge of parameters such as on or off rates, using an estimate of the wild-type interaction as the sole input. Utilizing relative steady-state responses, our protocol also allows for the rapid, reliable, and simultaneous determination of protein-binding dissociation constants of a large series of DNA mutants in a single experiment in a semiquantitative fashion. We compare our approach to existing methods, highlighting specific advantages as well as limitations. © 2013 Elsevier Inc. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Gamsjaeger, R. and Kariawasam, R. and Bang, L.H. and Touma, C. and Nguyen, C.D. and Matthews, J.M. and Cubeddu, L. and Mackay, J.P.\",\"doi\":\"10.1016/j.ab.2013.04.030\",\"journal\":\"Analytical Biochemistry\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Semiquantitative and quantitative analysis of protein-DNA interactions using steady-state measurements in surface plasmon resonance competition experiments},\\n type = {article},\\n year = {2013},\\n pages = {178-185},\\n volume = {440},\\n id = {cb78304b-2ce1-32bf-ae2e-25e278ca3b2c},\\n created = {2023-01-10T01:44:51.607Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:51.607Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {One method commonly used to characterize protein-DNA interactions is surface plasmon resonance (SPR). In a typical SPR experiment, chip-bound DNA is exposed to increasing concentrations of protein; the resulting binding data are used to calculate a dissociation constant for the interaction. However, in cases in which knowledge of the specificity of the interaction is required, a large set of DNA variants has to be tested; this is time consuming and costly, in part because of the requirement for multiple SPR chips. We have developed a new protocol that uses steady-state binding levels in SPR competition experiments to determine protein-binding dissociation constants for a set of DNA variants. This approach is rapid and straightforward and requires the use of only a single SPR chip. Additionally, in contrast to other methods, our approach does not require prior knowledge of parameters such as on or off rates, using an estimate of the wild-type interaction as the sole input. Utilizing relative steady-state responses, our protocol also allows for the rapid, reliable, and simultaneous determination of protein-binding dissociation constants of a large series of DNA mutants in a single experiment in a semiquantitative fashion. We compare our approach to existing methods, highlighting specific advantages as well as limitations. © 2013 Elsevier Inc. All rights reserved.},\\n bibtype = {article},\\n author = {Gamsjaeger, R. and Kariawasam, R. and Bang, L.H. and Touma, C. and Nguyen, C.D. and Matthews, J.M. and Cubeddu, L. and Mackay, J.P.},\\n doi = {10.1016/j.ab.2013.04.030},\\n journal = {Analytical Biochemistry},\\n number = {2}\\n}\",\"author_short\":[\"Gamsjaeger,  R.\",\"Kariawasam,  R.\",\"Bang,  L.\",\"Touma,  C.\",\"Nguyen,  C.\",\"Matthews,  J.\",\"Cubeddu,  L.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gamsjaeger-kariawasam-bang-touma-nguyen-matthews-cubeddu-mackay-semiquantitativeandquantitativeanalysisofproteindnainteractionsusingsteadystatemeasurementsinsurfaceplasmonresonancecompetitionexperiments-2013\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Analysis of disease-causing GATA1 mutations in murine gene complementation systems\",\"type\":\"article\",\"year\":\"2013\",\"pages\":\"5218-5227\",\"volume\":\"121\",\"id\":\"ddfb0231-026d-32b5-a980-22e8754a9d8b\",\"created\":\"2023-01-10T01:44:52.681Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:52.681Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Missense mutations in transcription factor GATA1 underlie a spectrum of congenital red blood cell and platelet disorders. We investigated how these alterations cause distinct clinical phenotypes by combining structural, biochemical, and genomic approaches with gene complementation systems that examine GATA1 function in biologically relevant cellular contexts. Substitutions that disrupt FOG1 cofactor binding impair both gene activation and repression and are associated with pronounced clinical phenotypes. Moreover, clinical severity correlates with the degree of FOG1 disruption. Surprisingly, 2 mutations shown to impair DNA binding of GATA1 in vitro did not measurably affect in vivo target gene occupancy. Rather, one of these disrupted binding to the TAL1 complex, implicating it in diseases caused by GATA1 mutations. Diminished TAL1 complex recruitment mainly impairs transcriptional activation and is linked to relatively mild disease. Notably, different substitutions at the same amino acid can selectively inhibit TAL1 complex or FOG1 binding, producing distinct cellular and clinical phenotypes. The structure-function relationships elucidated here were not predicted by prior in vitro or computational studies. Thus, our findings uncover novel disease mechanisms underlying GATA1 mutations and highlight the power of gene complementation assays for elucidating the molecular basis of genetic diseases.\",\"bibtype\":\"article\",\"author\":\"Campbell, A.E. and Wilkinson-White, L. and MacKay, J.P. and Matthews, J.M. and Blobel, G.A.\",\"doi\":\"10.1182/blood-2013-03-488080\",\"journal\":\"Blood\",\"number\":\"26\",\"bibtex\":\"@article{\\n title = {Analysis of disease-causing GATA1 mutations in murine gene complementation systems},\\n type = {article},\\n year = {2013},\\n pages = {5218-5227},\\n volume = {121},\\n id = {ddfb0231-026d-32b5-a980-22e8754a9d8b},\\n created = {2023-01-10T01:44:52.681Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:52.681Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Missense mutations in transcription factor GATA1 underlie a spectrum of congenital red blood cell and platelet disorders. We investigated how these alterations cause distinct clinical phenotypes by combining structural, biochemical, and genomic approaches with gene complementation systems that examine GATA1 function in biologically relevant cellular contexts. Substitutions that disrupt FOG1 cofactor binding impair both gene activation and repression and are associated with pronounced clinical phenotypes. Moreover, clinical severity correlates with the degree of FOG1 disruption. Surprisingly, 2 mutations shown to impair DNA binding of GATA1 in vitro did not measurably affect in vivo target gene occupancy. Rather, one of these disrupted binding to the TAL1 complex, implicating it in diseases caused by GATA1 mutations. Diminished TAL1 complex recruitment mainly impairs transcriptional activation and is linked to relatively mild disease. Notably, different substitutions at the same amino acid can selectively inhibit TAL1 complex or FOG1 binding, producing distinct cellular and clinical phenotypes. The structure-function relationships elucidated here were not predicted by prior in vitro or computational studies. Thus, our findings uncover novel disease mechanisms underlying GATA1 mutations and highlight the power of gene complementation assays for elucidating the molecular basis of genetic diseases.},\\n bibtype = {article},\\n author = {Campbell, A.E. and Wilkinson-White, L. and MacKay, J.P. and Matthews, J.M. and Blobel, G.A.},\\n doi = {10.1182/blood-2013-03-488080},\\n journal = {Blood},\\n number = {26}\\n}\",\"author_short\":[\"Campbell,  A.\",\"Wilkinson-White,  L.\",\"MacKay,  J.\",\"Matthews,  J.\",\"Blobel,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"campbell-wilkinsonwhite-mackay-matthews-blobel-analysisofdiseasecausinggata1mutationsinmurinegenecomplementationsystems-2013\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution structure of the LIM-homeodomain transcription factor complex Lhx3/Ldb1 and the effects of a pituitary mutation on key Lhx3 interactions\",\"type\":\"article\",\"year\":\"2012\",\"volume\":\"7\",\"id\":\"128d3304-f755-382e-92e5-6fc2f2229b4f\",\"created\":\"2023-01-10T01:44:53.740Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:53.740Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Lhx3 is a LIM-homeodomain (LIM-HD) transcription factor that regulates neural cell subtype specification and pituitary development in vertebrates, and mutations in this protein cause combined pituitary hormone deficiency syndrome (CPHDS). The recently published structures of Lhx3 in complex with each of two key protein partners, Isl1 and Ldb1, provide an opportunity to understand the effect of mutations and posttranslational modifications on key protein-protein interactions. Here, we use small-angle X-ray scattering of an Ldb1-Lhx3 complex to confirm that in solution the protein is well represented by our previously determined NMR structure as an ensemble of conformers each comprising two well-defined halves (each made up of LIM domain from Lhx3 and the corresponding binding motif in Ldb1) with some flexibility between the two halves. NMR analysis of an Lhx3 mutant that causes CPHDS, Lhx3(Y114C), shows that the mutation does not alter the zinc-ligation properties of Lhx3, but appears to cause a structural rearrangement of the hydrophobic core of the LIM2 domain of Lhx3 that destabilises the domain and/or reduces the affinity of Lhx3 for both Ldb1 and Isl1. Thus the mutation would affect the formation of Lhx3-containing transcription factor complexes, particularly in the pituitary gland where these complexes are required for the production of multiple pituitary cell types and hormones. © 2012 Bhati et al.\",\"bibtype\":\"article\",\"author\":\"Bhati, M. and Lee, C. and Gadd, M.S. and Jeffries, C.M. and Kwan, A. and Whitten, A.E. and Trewhella, J. and Mackay, J.P. and Matthews, J.M.\",\"doi\":\"10.1371/journal.pone.0040719\",\"journal\":\"PLoS ONE\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {Solution structure of the LIM-homeodomain transcription factor complex Lhx3/Ldb1 and the effects of a pituitary mutation on key Lhx3 interactions},\\n type = {article},\\n year = {2012},\\n volume = {7},\\n id = {128d3304-f755-382e-92e5-6fc2f2229b4f},\\n created = {2023-01-10T01:44:53.740Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:53.740Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Lhx3 is a LIM-homeodomain (LIM-HD) transcription factor that regulates neural cell subtype specification and pituitary development in vertebrates, and mutations in this protein cause combined pituitary hormone deficiency syndrome (CPHDS). The recently published structures of Lhx3 in complex with each of two key protein partners, Isl1 and Ldb1, provide an opportunity to understand the effect of mutations and posttranslational modifications on key protein-protein interactions. Here, we use small-angle X-ray scattering of an Ldb1-Lhx3 complex to confirm that in solution the protein is well represented by our previously determined NMR structure as an ensemble of conformers each comprising two well-defined halves (each made up of LIM domain from Lhx3 and the corresponding binding motif in Ldb1) with some flexibility between the two halves. NMR analysis of an Lhx3 mutant that causes CPHDS, Lhx3(Y114C), shows that the mutation does not alter the zinc-ligation properties of Lhx3, but appears to cause a structural rearrangement of the hydrophobic core of the LIM2 domain of Lhx3 that destabilises the domain and/or reduces the affinity of Lhx3 for both Ldb1 and Isl1. Thus the mutation would affect the formation of Lhx3-containing transcription factor complexes, particularly in the pituitary gland where these complexes are required for the production of multiple pituitary cell types and hormones. © 2012 Bhati et al.},\\n bibtype = {article},\\n author = {Bhati, M. and Lee, C. and Gadd, M.S. and Jeffries, C.M. and Kwan, A. and Whitten, A.E. and Trewhella, J. and Mackay, J.P. and Matthews, J.M.},\\n doi = {10.1371/journal.pone.0040719},\\n journal = {PLoS ONE},\\n number = {7}\\n}\",\"author_short\":[\"Bhati,  M.\",\"Lee,  C.\",\"Gadd,  M.\",\"Jeffries,  C.\",\"Kwan,  A.\",\"Whitten,  A.\",\"Trewhella,  J.\",\"Mackay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"bhati-lee-gadd-jeffries-kwan-whitten-trewhella-mackay-etal-solutionstructureofthelimhomeodomaintranscriptionfactorcomplexlhx3ldb1andtheeffectsofapituitarymutationonkeylhx3interactions-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A rapid method for assessing the RNA-binding potential of a protein\",\"type\":\"article\",\"year\":\"2012\",\"volume\":\"40\",\"id\":\"45db7a3c-9c0b-3ecb-a0b6-8ca4c8c9c8cc\",\"created\":\"2023-01-10T01:44:54.656Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:54.656Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures. © The Author(s) 2012.\",\"bibtype\":\"article\",\"author\":\"Bendak, K. and Loughlin, F.E. and Cheung, V. and O'Connell, M.R. and Crossley, M. and MacKay, J.P.\",\"doi\":\"10.1093/nar/gks285\",\"journal\":\"Nucleic Acids Research\",\"number\":\"14\",\"bibtex\":\"@article{\\n title = {A rapid method for assessing the RNA-binding potential of a protein},\\n type = {article},\\n year = {2012},\\n volume = {40},\\n id = {45db7a3c-9c0b-3ecb-a0b6-8ca4c8c9c8cc},\\n created = {2023-01-10T01:44:54.656Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:54.656Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures. © The Author(s) 2012.},\\n bibtype = {article},\\n author = {Bendak, K. and Loughlin, F.E. and Cheung, V. and O'Connell, M.R. and Crossley, M. and MacKay, J.P.},\\n doi = {10.1093/nar/gks285},\\n journal = {Nucleic Acids Research},\\n number = {14}\\n}\",\"author_short\":[\"Bendak,  K.\",\"Loughlin,  F.\",\"Cheung,  V.\",\"O'Connell,  M.\",\"Crossley,  M.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"bendak-loughlin-cheung-oconnell-crossley-mackay-arapidmethodforassessingthernabindingpotentialofaprotein-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Bivalent recognition of nucleosomes by the tandem PHD fingers of the CHD4 ATPase is required for CHD4-mediated repression\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"787-792\",\"volume\":\"109\",\"id\":\"fd6d1a67-ad29-356c-9c60-bd33f657e985\",\"created\":\"2023-01-10T01:44:55.575Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:55.575Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"CHD4 is a catalytic subunit of the NuRD (nucleosome remodeling and deacetylase) complex essential in transcriptional regulation, chromatin assembly and DNA damage repair. CHD4 contains tandem plant homeodomain (PHD) fingers connected by a short linker, the biological function of which remains unclear. Here we explore the combinatorial action of the CHD4 PHD1/2 fingers and detail the molecular basis for their association with chromatin. We found that PHD1/2 targets nucleosomes in a multivalent manner, concomitantly engaging two histone H3 tails. This robust synergistic interaction displaces HP1γ from pericentric sites, inducing changes in chromatin structure and leading to the dispersion of the heterochromatic mark H3K9me3. We demonstrate that recognition of the histone H3 tails by the PHD fingers is required for repressive activity of the CHD4/NuRD complex. Together, our data elucidate the molecular mechanism of multivalent association of the PHD fingers with chromatin and reveal their critical role in the regulation of CHD4 functions.\",\"bibtype\":\"article\",\"author\":\"Musselman, C.A. and Ramiŕez, J. and Sims, J.K. and Mansfield, R.E. and Oliver, S.S. and Denu, J.M. and Mackay, J.P. and Wade, P.A. and Hagman, J. and Kutateladze, T.G.\",\"doi\":\"10.1073/pnas.1113655109\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Bivalent recognition of nucleosomes by the tandem PHD fingers of the CHD4 ATPase is required for CHD4-mediated repression},\\n type = {article},\\n year = {2012},\\n pages = {787-792},\\n volume = {109},\\n id = {fd6d1a67-ad29-356c-9c60-bd33f657e985},\\n created = {2023-01-10T01:44:55.575Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:55.575Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {CHD4 is a catalytic subunit of the NuRD (nucleosome remodeling and deacetylase) complex essential in transcriptional regulation, chromatin assembly and DNA damage repair. CHD4 contains tandem plant homeodomain (PHD) fingers connected by a short linker, the biological function of which remains unclear. Here we explore the combinatorial action of the CHD4 PHD1/2 fingers and detail the molecular basis for their association with chromatin. We found that PHD1/2 targets nucleosomes in a multivalent manner, concomitantly engaging two histone H3 tails. This robust synergistic interaction displaces HP1γ from pericentric sites, inducing changes in chromatin structure and leading to the dispersion of the heterochromatic mark H3K9me3. We demonstrate that recognition of the histone H3 tails by the PHD fingers is required for repressive activity of the CHD4/NuRD complex. Together, our data elucidate the molecular mechanism of multivalent association of the PHD fingers with chromatin and reveal their critical role in the regulation of CHD4 functions.},\\n bibtype = {article},\\n author = {Musselman, C.A. and Ramiŕez, J. and Sims, J.K. and Mansfield, R.E. and Oliver, S.S. and Denu, J.M. and Mackay, J.P. and Wade, P.A. and Hagman, J. and Kutateladze, T.G.},\\n doi = {10.1073/pnas.1113655109},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {3}\\n}\",\"author_short\":[\"Musselman,  C.\",\"Ramiŕez,  J.\",\"Sims,  J.\",\"Mansfield,  R.\",\"Oliver,  S.\",\"Denu,  J.\",\"Mackay,  J.\",\"Wade,  P.\",\"Hagman,  J.\",\"Kutateladze,  T.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"musselman-ramiez-sims-mansfield-oliver-denu-mackay-wade-etal-bivalentrecognitionofnucleosomesbythetandemphdfingersofthechd4atpaseisrequiredforchd4mediatedrepression-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Two-timing zinc finger transcription factors liaising with RNA\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"199-205\",\"volume\":\"37\",\"id\":\"2554db33-7b4b-3127-99cf-3fde4951ed45\",\"created\":\"2023-01-10T01:44:56.496Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:56.496Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Classical zinc fingers (ZFs) are one of the most common protein domains in higher eukaryotes and have been known for almost 30 years to act as sequence-specific DNA-binding domains. This knowledge has come, however, from the study of a small number of archetypal proteins, and a larger picture is beginning to emerge that ZF functions are far more diverse than originally suspected. Here, we review the evidence that a subset of ZF proteins live double lives, binding to both DNA and RNA targets and frequenting both the cytoplasm and the nucleus. This duality can create an important additional level of gene regulation that serves to connect transcriptional and post-transcriptional control. © 2012 Elsevier Ltd.\",\"bibtype\":\"article\",\"author\":\"Burdach, J. and O'Connell, M.R. and Mackay, J.P. and Crossley, M.\",\"doi\":\"10.1016/j.tibs.2012.02.001\",\"journal\":\"Trends in Biochemical Sciences\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Two-timing zinc finger transcription factors liaising with RNA},\\n type = {article},\\n year = {2012},\\n pages = {199-205},\\n volume = {37},\\n id = {2554db33-7b4b-3127-99cf-3fde4951ed45},\\n created = {2023-01-10T01:44:56.496Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:56.496Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Classical zinc fingers (ZFs) are one of the most common protein domains in higher eukaryotes and have been known for almost 30 years to act as sequence-specific DNA-binding domains. This knowledge has come, however, from the study of a small number of archetypal proteins, and a larger picture is beginning to emerge that ZF functions are far more diverse than originally suspected. Here, we review the evidence that a subset of ZF proteins live double lives, binding to both DNA and RNA targets and frequenting both the cytoplasm and the nucleus. This duality can create an important additional level of gene regulation that serves to connect transcriptional and post-transcriptional control. © 2012 Elsevier Ltd.},\\n bibtype = {article},\\n author = {Burdach, J. and O'Connell, M.R. and Mackay, J.P. and Crossley, M.},\\n doi = {10.1016/j.tibs.2012.02.001},\\n journal = {Trends in Biochemical Sciences},\\n number = {5}\\n}\",\"author_short\":[\"Burdach,  J.\",\"O'Connell,  M.\",\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"burdach-oconnell-mackay-crossley-twotimingzincfingertranscriptionfactorsliaisingwithrna-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution structure of a tethered Lmo2<inf>LIM2</inf>/Ldb1<inf>LID</inf> complex\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"1768-1774\",\"volume\":\"21\",\"id\":\"c8d8b2ce-e804-3b1e-bc6d-dea5554342b7\",\"created\":\"2023-01-10T01:44:57.390Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:57.390Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.\",\"bibtype\":\"article\",\"author\":\"Dastmalchi, S. and Wilkinson-White, L. and Kwan, A.H. and Gamsjaeger, R. and Mackay, J.P. and Matthews, J.M.\",\"doi\":\"10.1002/pro.2153\",\"journal\":\"Protein Science\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Solution structure of a tethered Lmo2<inf>LIM2</inf>/Ldb1<inf>LID</inf> complex},\\n type = {article},\\n year = {2012},\\n pages = {1768-1774},\\n volume = {21},\\n id = {c8d8b2ce-e804-3b1e-bc6d-dea5554342b7},\\n created = {2023-01-10T01:44:57.390Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:57.390Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.},\\n bibtype = {article},\\n author = {Dastmalchi, S. and Wilkinson-White, L. and Kwan, A.H. and Gamsjaeger, R. and Mackay, J.P. and Matthews, J.M.},\\n doi = {10.1002/pro.2153},\\n journal = {Protein Science},\\n number = {11}\\n}\",\"author_short\":[\"Dastmalchi,  S.\",\"Wilkinson-White,  L.\",\"Kwan,  A.\",\"Gamsjaeger,  R.\",\"Mackay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2inflim2infldb1inflidinfcomplex-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Modular assembly of RanBP2-Type zinc finger domains to target single-stranded RNA\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"5371-5375\",\"volume\":\"51\",\"id\":\"524d43b0-eac3-303b-9076-d08253f97d3b\",\"created\":\"2023-01-10T01:44:58.301Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:58.301Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Fingering RNA: To study the function of the variety of RNA species that have been discovered recently, sequence-specific RNA-binding molecules with tunable specificity are required. Here, the use of zinc fingers (ZFs) from the splicing factors ZRANB2 and RBM5 as potential modules for the assembly of sequence-specific RNA-binding molecules is demonstrated. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.\",\"bibtype\":\"article\",\"author\":\"O'Connell, M.R. and Vandevenne, M. and Nguyen, C.D. and Matthews, J.M. and Gamsjaeger, R. and Segal, D.J. and MacKay, J.P.\",\"doi\":\"10.1002/anie.201200866\",\"journal\":\"Angewandte Chemie - International Edition\",\"number\":\"22\",\"bibtex\":\"@article{\\n title = {Modular assembly of RanBP2-Type zinc finger domains to target single-stranded RNA},\\n type = {article},\\n year = {2012},\\n pages = {5371-5375},\\n volume = {51},\\n id = {524d43b0-eac3-303b-9076-d08253f97d3b},\\n created = {2023-01-10T01:44:58.301Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:58.301Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Fingering RNA: To study the function of the variety of RNA species that have been discovered recently, sequence-specific RNA-binding molecules with tunable specificity are required. Here, the use of zinc fingers (ZFs) from the splicing factors ZRANB2 and RBM5 as potential modules for the assembly of sequence-specific RNA-binding molecules is demonstrated. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.},\\n bibtype = {article},\\n author = {O'Connell, M.R. and Vandevenne, M. and Nguyen, C.D. and Matthews, J.M. and Gamsjaeger, R. and Segal, D.J. and MacKay, J.P.},\\n doi = {10.1002/anie.201200866},\\n journal = {Angewandte Chemie - International Edition},\\n number = {22}\\n}\",\"author_short\":[\"O'Connell,  M.\",\"Vandevenne,  M.\",\"Nguyen,  C.\",\"Matthews,  J.\",\"Gamsjaeger,  R.\",\"Segal,  D.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"oconnell-vandevenne-nguyen-matthews-gamsjaeger-segal-mackay-modularassemblyofranbp2typezincfingerdomainstotargetsinglestrandedrna-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular LMO4-LIM1/CtIP complex\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"31-34\",\"volume\":\"6\",\"id\":\"f121ae58-5c4e-3fbb-987c-35bd8fe224e1\",\"created\":\"2023-01-10T01:44:59.248Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:44:59.248Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"LMO4 is a broadly expressed LIM-only protein that is involved in neural tube development and implicated in breast cancer. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the N-terminal LIM domain from LMO4 tethered to residues 641-685 of C-terminal binding protein interacting protein (CtIP/RBBP8). © 2011 Springer Science+Business Media B.V.\",\"bibtype\":\"article\",\"author\":\"Liew, C.W. and Kwan, A.H. and Stokes, P.H. and MacKay, J.P. and Matthews, J.M.\",\"doi\":\"10.1007/s12104-011-9319-0\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular LMO4-LIM1/CtIP complex},\\n type = {article},\\n year = {2012},\\n pages = {31-34},\\n volume = {6},\\n id = {f121ae58-5c4e-3fbb-987c-35bd8fe224e1},\\n created = {2023-01-10T01:44:59.248Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:44:59.248Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {LMO4 is a broadly expressed LIM-only protein that is involved in neural tube development and implicated in breast cancer. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the N-terminal LIM domain from LMO4 tethered to residues 641-685 of C-terminal binding protein interacting protein (CtIP/RBBP8). © 2011 Springer Science+Business Media B.V.},\\n bibtype = {article},\\n author = {Liew, C.W. and Kwan, A.H. and Stokes, P.H. and MacKay, J.P. and Matthews, J.M.},\\n doi = {10.1007/s12104-011-9319-0},\\n journal = {Biomolecular NMR Assignments},\\n number = {1}\\n}\",\"author_short\":[\"Liew,  C.\",\"Kwan,  A.\",\"Stokes,  P.\",\"MacKay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"liew-kwan-stokes-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo4lim1ctipcomplex-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Synthesis of the bacteriocin glycopeptide sublancin 168 and S-glycosylated variants\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"1910-1913\",\"volume\":\"14\",\"id\":\"0782aeb9-904a-31dc-bf32-5d68b28b7eb4\",\"created\":\"2023-01-10T01:45:00.240Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:00.240Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The synthesis of sublancin 168, a unique S-glucosylated bacteriocin antibiotic, is described. The natural product and two S-glycosylated variants were successfully prepared via native chemical ligation followed by folding. The synthetic glycopeptides were shown to possess primarily an α-helical secondary structure by CD and NMR studies. © 2012 American Chemical Society.\",\"bibtype\":\"article\",\"author\":\"Hsieh, Y.S.Y. and Wilkinson, B.L. and O'Connell, M.R. and MacKay, J.P. and Matthews, J.M. and Payne, R.J.\",\"doi\":\"10.1021/ol300557g\",\"journal\":\"Organic Letters\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {Synthesis of the bacteriocin glycopeptide sublancin 168 and S-glycosylated variants},\\n type = {article},\\n year = {2012},\\n pages = {1910-1913},\\n volume = {14},\\n id = {0782aeb9-904a-31dc-bf32-5d68b28b7eb4},\\n created = {2023-01-10T01:45:00.240Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:00.240Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The synthesis of sublancin 168, a unique S-glucosylated bacteriocin antibiotic, is described. The natural product and two S-glycosylated variants were successfully prepared via native chemical ligation followed by folding. The synthetic glycopeptides were shown to possess primarily an α-helical secondary structure by CD and NMR studies. © 2012 American Chemical Society.},\\n bibtype = {article},\\n author = {Hsieh, Y.S.Y. and Wilkinson, B.L. and O'Connell, M.R. and MacKay, J.P. and Matthews, J.M. and Payne, R.J.},\\n doi = {10.1021/ol300557g},\\n journal = {Organic Letters},\\n number = {7}\\n}\",\"author_short\":[\"Hsieh,  Y.\",\"Wilkinson,  B.\",\"O'Connell,  M.\",\"MacKay,  J.\",\"Matthews,  J.\",\"Payne,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"hsieh-wilkinson-oconnell-mackay-matthews-payne-synthesisofthebacteriocinglycopeptidesublancin168andsglycosylatedvariants-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Backbone and sidechain <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"83-86\",\"volume\":\"6\",\"id\":\"ff9a844c-9cab-30ff-b896-a298653e06c7\",\"created\":\"2023-01-10T01:45:01.153Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:01.153Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.\",\"bibtype\":\"article\",\"author\":\"Morris, V.K. and Kwan, A.H. and MacKay, J.P. and Sunde, M.\",\"doi\":\"10.1007/s12104-011-9330-5\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Backbone and sidechain <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans},\\n type = {article},\\n year = {2012},\\n pages = {83-86},\\n volume = {6},\\n id = {ff9a844c-9cab-30ff-b896-a298653e06c7},\\n created = {2023-01-10T01:45:01.153Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:01.153Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.},\\n bibtype = {article},\\n author = {Morris, V.K. and Kwan, A.H. and MacKay, J.P. and Sunde, M.},\\n doi = {10.1007/s12104-011-9330-5},\\n journal = {Biomolecular NMR Assignments},\\n number = {1}\\n}\",\"author_short\":[\"Morris,  V.\",\"Kwan,  A.\",\"MacKay,  J.\",\"Sunde,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"morris-kwan-mackay-sunde-backboneandsidechainsup1suphsup13supcandsup15supnchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"1267-1278\",\"volume\":\"18\",\"id\":\"620041f4-f32a-3709-9ae9-50c212e6b3e8\",\"created\":\"2023-01-10T01:45:02.072Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:02.072Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.\",\"bibtype\":\"article\",\"author\":\"Mckenzie, J.L. and Duyvestyn, J.M. and Smith, T. and Bendak, K. and Mackay, J. and Cursons, R.A.Y. and Cook, G.M. and Arcus, V.L.\",\"doi\":\"10.1261/rna.031229.111\",\"journal\":\"RNA\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry},\\n type = {article},\\n year = {2012},\\n pages = {1267-1278},\\n volume = {18},\\n id = {620041f4-f32a-3709-9ae9-50c212e6b3e8},\\n created = {2023-01-10T01:45:02.072Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:02.072Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.},\\n bibtype = {article},\\n author = {Mckenzie, J.L. and Duyvestyn, J.M. and Smith, T. and Bendak, K. and Mackay, J. and Cursons, R.A.Y. and Cook, G.M. and Arcus, V.L.},\\n doi = {10.1261/rna.031229.111},\\n journal = {RNA},\\n number = {6}\\n}\",\"author_short\":[\"Mckenzie,  J.\",\"Duyvestyn,  J.\",\"Smith,  T.\",\"Bendak,  K.\",\"Mackay,  J.\",\"Cursons,  R.\",\"Cook,  G.\",\"Arcus,  V.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Interdomain orientation of cardiac troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state\",\"type\":\"article\",\"year\":\"2012\",\"pages\":\"1376-1387\",\"volume\":\"21\",\"id\":\"4e767ca7-1e6a-3aae-aa86-c051dd322f85\",\"created\":\"2023-01-10T01:45:03.028Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:03.028Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.\",\"bibtype\":\"article\",\"author\":\"Cordina, N.M. and Liew, C.K. and Gell, D.A. and Fajer, P.G. and Mackay, J.P. and Brown, L.J.\",\"doi\":\"10.1002/pro.2124\",\"journal\":\"Protein Science\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {Interdomain orientation of cardiac troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state},\\n type = {article},\\n year = {2012},\\n pages = {1376-1387},\\n volume = {21},\\n id = {4e767ca7-1e6a-3aae-aa86-c051dd322f85},\\n created = {2023-01-10T01:45:03.028Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:03.028Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.},\\n bibtype = {article},\\n author = {Cordina, N.M. and Liew, C.K. and Gell, D.A. and Fajer, P.G. and Mackay, J.P. and Brown, L.J.},\\n doi = {10.1002/pro.2124},\\n journal = {Protein Science},\\n number = {9}\\n}\",\"author_short\":[\"Cordina,  N.\",\"Liew,  C.\",\"Gell,  D.\",\"Fajer,  P.\",\"Mackay,  J.\",\"Brown,  L.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Self-assembly of functional, amphipathic amyloid monolayers by the fungal hydrophobin EAS\",\"type\":\"article\",\"year\":\"2012\",\"volume\":\"109\",\"id\":\"b585d9ee-1807-3f19-9891-d5c72141f251\",\"created\":\"2023-01-10T01:45:04.060Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:04.060Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The hydrophobin EAS from the fungus Neurospora crassa forms functional amyloid fibrils called rodlets that facilitate spore formation and dispersal. Self-assembly of EAS into fibrillar rodlets occurs spontaneously at hydrophobic:hydrophilic interfaces and the rodlets further associate laterally to form amphipathic monolayers. We have used site-directed mutagenesis and peptide experiments to identify the region of EAS that drives intermolecular association and formation of the cross-β rodlet structure. Transplanting this region into a nonamyloidogenic hydrophobin enables it to form rodlets. We have also determined the structure and dynamics of an EAS variant with reduced rodlet-forming ability. Taken together, these data allow us to pinpoint the conformational changes that take place when hydrophobins self-assemble at an interface and to propose a model for the amphipathic EAS rodlet structure.\",\"bibtype\":\"article\",\"author\":\"Macindoe, I. and Kwan, A.H. and Ren, Q. and Morris, V.K. and Yang, W. and Mackay, J.P. and Sunde, M.\",\"doi\":\"10.1073/pnas.1114052109\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"14\",\"bibtex\":\"@article{\\n title = {Self-assembly of functional, amphipathic amyloid monolayers by the fungal hydrophobin EAS},\\n type = {article},\\n year = {2012},\\n volume = {109},\\n id = {b585d9ee-1807-3f19-9891-d5c72141f251},\\n created = {2023-01-10T01:45:04.060Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:04.060Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The hydrophobin EAS from the fungus Neurospora crassa forms functional amyloid fibrils called rodlets that facilitate spore formation and dispersal. Self-assembly of EAS into fibrillar rodlets occurs spontaneously at hydrophobic:hydrophilic interfaces and the rodlets further associate laterally to form amphipathic monolayers. We have used site-directed mutagenesis and peptide experiments to identify the region of EAS that drives intermolecular association and formation of the cross-β rodlet structure. Transplanting this region into a nonamyloidogenic hydrophobin enables it to form rodlets. We have also determined the structure and dynamics of an EAS variant with reduced rodlet-forming ability. Taken together, these data allow us to pinpoint the conformational changes that take place when hydrophobins self-assemble at an interface and to propose a model for the amphipathic EAS rodlet structure.},\\n bibtype = {article},\\n author = {Macindoe, I. and Kwan, A.H. and Ren, Q. and Morris, V.K. and Yang, W. and Mackay, J.P. and Sunde, M.},\\n doi = {10.1073/pnas.1114052109},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {14}\\n}\",\"author_short\":[\"Macindoe,  I.\",\"Kwan,  A.\",\"Ren,  Q.\",\"Morris,  V.\",\"Yang,  W.\",\"Mackay,  J.\",\"Sunde,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"macindoe-kwan-ren-morris-yang-mackay-sunde-selfassemblyoffunctionalamphipathicamyloidmonolayersbythefungalhydrophobineas-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The detection and quantitation of protein oligomerization\",\"type\":\"book\",\"year\":\"2012\",\"source\":\"Advances in Experimental Medicine and Biology\",\"pages\":\"19-41\",\"volume\":\"747\",\"id\":\"7e3e3789-bc90-34ed-805a-f49344268f6e\",\"created\":\"2023-01-10T01:45:04.982Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:04.982Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"There are many different techniques available to biologists and biochemists that can be used to detect and characterize the self-association of proteins. Each technique has strengths and weaknesses and it is often useful to combine several approaches to maximize the former and minimize the latter. Here we review a range of methodologies that identify protein self-association and/or allow the stoichiometry and affinity of the interaction to be determined, placing an emphasis on what type of information can be obtained and outlining the advantages and disadvantages involved. In general, in vitro biophysical techniques, such as size exclusion chromatography, analytical ultracentrifugation, scattering techniques, NMR spectroscopy, isothermal titration calorimetry, fluorescence anisotropy and mass spectrometry, provide information on stoichiometry and/or binding affinities. Other approaches such as cross-linking, fluorescence methods (e.g., fluorescence correlation spectroscopy, FCS; Förster resonance energy transfer, FRET; fluorescence recovery after photobleaching, FRAP; and proximity imaging, PRIM) and complementation approaches (e.g., yeast two hybrid assays and bimolecular fluorescence complementation, BiFC) can be used to detect protein self-association in a cellular context. © 2012 Springer Science+Business Media, LLC.\",\"bibtype\":\"book\",\"author\":\"Gell, D.A. and Grant, R.P. and MacKay, J.P.\",\"doi\":\"10.1007/978-1-4614-3229-6_2\",\"bibtex\":\"@book{\\n title = {The detection and quantitation of protein oligomerization},\\n type = {book},\\n year = {2012},\\n source = {Advances in Experimental Medicine and Biology},\\n pages = {19-41},\\n volume = {747},\\n id = {7e3e3789-bc90-34ed-805a-f49344268f6e},\\n created = {2023-01-10T01:45:04.982Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:04.982Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {There are many different techniques available to biologists and biochemists that can be used to detect and characterize the self-association of proteins. Each technique has strengths and weaknesses and it is often useful to combine several approaches to maximize the former and minimize the latter. Here we review a range of methodologies that identify protein self-association and/or allow the stoichiometry and affinity of the interaction to be determined, placing an emphasis on what type of information can be obtained and outlining the advantages and disadvantages involved. In general, in vitro biophysical techniques, such as size exclusion chromatography, analytical ultracentrifugation, scattering techniques, NMR spectroscopy, isothermal titration calorimetry, fluorescence anisotropy and mass spectrometry, provide information on stoichiometry and/or binding affinities. Other approaches such as cross-linking, fluorescence methods (e.g., fluorescence correlation spectroscopy, FCS; Förster resonance energy transfer, FRET; fluorescence recovery after photobleaching, FRAP; and proximity imaging, PRIM) and complementation approaches (e.g., yeast two hybrid assays and bimolecular fluorescence complementation, BiFC) can be used to detect protein self-association in a cellular context. © 2012 Springer Science+Business Media, LLC.},\\n bibtype = {book},\\n author = {Gell, D.A. and Grant, R.P. and MacKay, J.P.},\\n doi = {10.1007/978-1-4614-3229-6_2}\\n}\",\"author_short\":[\"Gell,  D.\",\"Grant,  R.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gell-grant-mackay-thedetectionandquantitationofproteinoligomerization-2012\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The multi-zinc finger protein ZNF217 contacts DNA through a two-finger domain\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"38190-38201\",\"volume\":\"286\",\"id\":\"e1fdc7f2-dcf3-344a-b417-57036c8d3fcc\",\"created\":\"2023-01-10T01:45:05.935Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:05.935Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Classical C2H2 zinc finger proteins are among the most abundant transcription factors found in eukaryotes, and the mechanisms through which they recognize their target genes have been extensively investigated. In general, a tandem array of three fingers separated by characteristic TGERP links is required for sequence-specific DNA recognition. Nevertheless, a significant number of zinc finger proteins do not contain a hallmark three-finger array of this type, raising the question of whether and how they contact DNA. We have examined the multi-finger protein ZNF217, which contains eight classical zinc fingers. ZNF217 is implicated as an oncogene and in repressing the E-cadherin gene. We show that two of its zinc fingers, 6 and 7, can mediate contacts with DNA. We examine its putative recognition site in the E-cadherin promoter and demonstrate that this is a suboptimal site. NMR analysis and mutagenesis is used to define the DNA binding surface of ZNF217, and we examine the specificity of the DNA binding activity using fluorescence anisotropy titrations. Finally, sequence analysis reveals that a variety of multi-finger proteins also contain two-finger units, and our data support the idea that these may constitute a distinct subclass of DNA recognition motif. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A.\",\"bibtype\":\"article\",\"author\":\"Nunez, N. and Clifton, M.M.K. and Funnell, A.P.W. and Artuz, C. and Hallal, S. and Quinlan, K.G.R. and Font, J. and Vandevenne, M. and Setiyaputra, S. and Pearson, R.C.M. and Mackay, J.P. and Crossley, M.\",\"doi\":\"10.1074/jbc.M111.301234\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"44\",\"bibtex\":\"@article{\\n title = {The multi-zinc finger protein ZNF217 contacts DNA through a two-finger domain},\\n type = {article},\\n year = {2011},\\n pages = {38190-38201},\\n volume = {286},\\n id = {e1fdc7f2-dcf3-344a-b417-57036c8d3fcc},\\n created = {2023-01-10T01:45:05.935Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:05.935Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Classical C2H2 zinc finger proteins are among the most abundant transcription factors found in eukaryotes, and the mechanisms through which they recognize their target genes have been extensively investigated. In general, a tandem array of three fingers separated by characteristic TGERP links is required for sequence-specific DNA recognition. Nevertheless, a significant number of zinc finger proteins do not contain a hallmark three-finger array of this type, raising the question of whether and how they contact DNA. We have examined the multi-finger protein ZNF217, which contains eight classical zinc fingers. ZNF217 is implicated as an oncogene and in repressing the E-cadherin gene. We show that two of its zinc fingers, 6 and 7, can mediate contacts with DNA. We examine its putative recognition site in the E-cadherin promoter and demonstrate that this is a suboptimal site. NMR analysis and mutagenesis is used to define the DNA binding surface of ZNF217, and we examine the specificity of the DNA binding activity using fluorescence anisotropy titrations. Finally, sequence analysis reveals that a variety of multi-finger proteins also contain two-finger units, and our data support the idea that these may constitute a distinct subclass of DNA recognition motif. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A.},\\n bibtype = {article},\\n author = {Nunez, N. and Clifton, M.M.K. and Funnell, A.P.W. and Artuz, C. and Hallal, S. and Quinlan, K.G.R. and Font, J. and Vandevenne, M. and Setiyaputra, S. and Pearson, R.C.M. and Mackay, J.P. and Crossley, M.},\\n doi = {10.1074/jbc.M111.301234},\\n journal = {Journal of Biological Chemistry},\\n number = {44}\\n}\",\"author_short\":[\"Nunez,  N.\",\"Clifton,  M.\",\"Funnell,  A.\",\"Artuz,  C.\",\"Hallal,  S.\",\"Quinlan,  K.\",\"Font,  J.\",\"Vandevenne,  M.\",\"Setiyaputra,  S.\",\"Pearson,  R.\",\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"nunez-clifton-funnell-artuz-hallal-quinlan-font-vandevenne-etal-themultizincfingerproteinznf217contactsdnathroughatwofingerdomain-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes\",\"type\":\"article\",\"year\":\"2011\",\"volume\":\"108\",\"id\":\"cd0868d1-a20c-3300-8452-75a3d9552903\",\"created\":\"2023-01-10T01:45:06.903Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:06.903Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 \\\"reads\\\" acetyl marks on nuclear factors to promote their stable association with chromatin.\",\"bibtype\":\"article\",\"author\":\"Lamonica, J.M. and Deng, W. and Kadauke, S. and Campbell, A.E. and Gamsjaeger, R. and Wang, H. and Cheng, Y. and Billin, A.N. and Hardison, R.C. and Mackay, J.P. and Mackay, J.P. and Blobel, G.A.\",\"doi\":\"10.1073/pnas.1102140108\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"22\",\"bibtex\":\"@article{\\n title = {Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes},\\n type = {article},\\n year = {2011},\\n volume = {108},\\n id = {cd0868d1-a20c-3300-8452-75a3d9552903},\\n created = {2023-01-10T01:45:06.903Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:06.903Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 \\\"reads\\\" acetyl marks on nuclear factors to promote their stable association with chromatin.},\\n bibtype = {article},\\n author = {Lamonica, J.M. and Deng, W. and Kadauke, S. and Campbell, A.E. and Gamsjaeger, R. and Wang, H. and Cheng, Y. and Billin, A.N. and Hardison, R.C. and Mackay, J.P. and Mackay, J.P. and Blobel, G.A.},\\n doi = {10.1073/pnas.1102140108},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {22}\\n}\",\"author_short\":[\"Lamonica,  J.\",\"Deng,  W.\",\"Kadauke,  S.\",\"Campbell,  A.\",\"Gamsjaeger,  R.\",\"Wang,  H.\",\"Cheng,  Y.\",\"Billin,  A.\",\"Hardison,  R.\",\"Mackay,  J.\",\"Mackay,  J.\",\"Blobel,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural basis for hemoglobin capture by Staphylococcus aureus cell-surface protein, IsdH\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"38439-38447\",\"volume\":\"286\",\"id\":\"206f34b7-fe09-3a3c-a885-250484075b13\",\"created\":\"2023-01-10T01:45:07.824Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:07.824Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Pathogens must steal iron from their hosts to establish infection. In mammals, hemoglobin (Hb) represents the largest reservoir of iron, and pathogens express Hb-binding proteins to access this source. Here, we show how one of the commonest and most significant human pathogens, Staphylococcus aureus, captures Hb as the first step of an iron-scavenging pathway. The x-ray crystal structure of Hb bound to a domain from the Isd (iron-regulated surface determinant) protein, IsdH, is the first structure of a Hb capture complex to be determined. Surface mutations in Hb that reduce binding to the Hb-receptor limit the capacity of S. aureus to utilize Hb as an iron source, suggesting that Hb sequence is a factor in host susceptibility to infection. The demonstration that pathogens make highly specific recognition complexes with Hb raises the possibility of developing inhibitors of Hb binding as antibacterial agents. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Kumar, K.K. and Jacques, D.A. and Pishchany, G. and Caradoc-Davies, T. and Spirig, T. and Malmirchegini, G.R. and Langley, D.B. and Dickson, C.F. and Mackay, J.P. and Clubb, R.T. and Guss, J.M. and Gell, D.A.\",\"doi\":\"10.1074/jbc.M111.287300\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"44\",\"bibtex\":\"@article{\\n title = {Structural basis for hemoglobin capture by Staphylococcus aureus cell-surface protein, IsdH},\\n type = {article},\\n year = {2011},\\n pages = {38439-38447},\\n volume = {286},\\n id = {206f34b7-fe09-3a3c-a885-250484075b13},\\n created = {2023-01-10T01:45:07.824Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:07.824Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Pathogens must steal iron from their hosts to establish infection. In mammals, hemoglobin (Hb) represents the largest reservoir of iron, and pathogens express Hb-binding proteins to access this source. Here, we show how one of the commonest and most significant human pathogens, Staphylococcus aureus, captures Hb as the first step of an iron-scavenging pathway. The x-ray crystal structure of Hb bound to a domain from the Isd (iron-regulated surface determinant) protein, IsdH, is the first structure of a Hb capture complex to be determined. Surface mutations in Hb that reduce binding to the Hb-receptor limit the capacity of S. aureus to utilize Hb as an iron source, suggesting that Hb sequence is a factor in host susceptibility to infection. The demonstration that pathogens make highly specific recognition complexes with Hb raises the possibility of developing inhibitors of Hb binding as antibacterial agents. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Kumar, K.K. and Jacques, D.A. and Pishchany, G. and Caradoc-Davies, T. and Spirig, T. and Malmirchegini, G.R. and Langley, D.B. and Dickson, C.F. and Mackay, J.P. and Clubb, R.T. and Guss, J.M. and Gell, D.A.},\\n doi = {10.1074/jbc.M111.287300},\\n journal = {Journal of Biological Chemistry},\\n number = {44}\\n}\",\"author_short\":[\"Kumar,  K.\",\"Jacques,  D.\",\"Pishchany,  G.\",\"Caradoc-Davies,  T.\",\"Spirig,  T.\",\"Malmirchegini,  G.\",\"Langley,  D.\",\"Dickson,  C.\",\"Mackay,  J.\",\"Clubb,  R.\",\"Guss,  J.\",\"Gell,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kumar-jacques-pishchany-caradocdavies-spirig-malmirchegini-langley-dickson-etal-structuralbasisforhemoglobincapturebystaphylococcusaureuscellsurfaceproteinisdh-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Protein-protein interactions: Analysis of a false positive GST pulldown result\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"2365-2371\",\"volume\":\"79\",\"id\":\"5d9c6e9b-c8ce-3c98-ab37-684192c58e66\",\"created\":\"2023-01-10T01:45:08.736Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:08.736Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"One of the most common ways to demonstrate a direct protein-protein interaction in vitro is the glutathione-S-transferse (GST)-pulldown. Here we report the detailed characterization of a putative interaction between two transcription factor proteins, GATA-1 and Krüppel-like factor 3 (KLF3/BKLF) that show robust interactions in GST-pulldown experiments. Attempts to map the interaction interface of GATA-1 on KLF3 using a mutagenic screening approach did not yield a contiguous binding face on KLF3, suggesting that the interaction might be non-specific. NMR experiments showed that the proteins do not interact at protein concentrations of 50-100 μM. Rather, the GST tag can cause part of KLF3 to misfold. In addition to misfolding, the fact that both proteins are DNA-binding domains appears to introduce binding artifacts (possibly nucleic acid bridging) that cannot be resolved by the addition of nucleases or ethidium bromide (EtBr). This study emphasizes the need for caution in relying on GST-pulldown results and related methods, without convincing confirmation from different approaches. © 2011 Wiley-Liss, Inc.\",\"bibtype\":\"article\",\"author\":\"Wissmueller, S. and Font, J. and Liew, C.W. and Cram, E. and Schroeder, T. and Turner, J. and Crossley, M. and Mackay, J.P. and Matthews, J.M.\",\"doi\":\"10.1002/prot.23068\",\"journal\":\"Proteins: Structure, Function and Bioinformatics\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {Protein-protein interactions: Analysis of a false positive GST pulldown result},\\n type = {article},\\n year = {2011},\\n pages = {2365-2371},\\n volume = {79},\\n id = {5d9c6e9b-c8ce-3c98-ab37-684192c58e66},\\n created = {2023-01-10T01:45:08.736Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:08.736Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {One of the most common ways to demonstrate a direct protein-protein interaction in vitro is the glutathione-S-transferse (GST)-pulldown. Here we report the detailed characterization of a putative interaction between two transcription factor proteins, GATA-1 and Krüppel-like factor 3 (KLF3/BKLF) that show robust interactions in GST-pulldown experiments. Attempts to map the interaction interface of GATA-1 on KLF3 using a mutagenic screening approach did not yield a contiguous binding face on KLF3, suggesting that the interaction might be non-specific. NMR experiments showed that the proteins do not interact at protein concentrations of 50-100 μM. Rather, the GST tag can cause part of KLF3 to misfold. In addition to misfolding, the fact that both proteins are DNA-binding domains appears to introduce binding artifacts (possibly nucleic acid bridging) that cannot be resolved by the addition of nucleases or ethidium bromide (EtBr). This study emphasizes the need for caution in relying on GST-pulldown results and related methods, without convincing confirmation from different approaches. © 2011 Wiley-Liss, Inc.},\\n bibtype = {article},\\n author = {Wissmueller, S. and Font, J. and Liew, C.W. and Cram, E. and Schroeder, T. and Turner, J. and Crossley, M. and Mackay, J.P. and Matthews, J.M.},\\n doi = {10.1002/prot.23068},\\n journal = {Proteins: Structure, Function and Bioinformatics},\\n number = {8}\\n}\",\"author_short\":[\"Wissmueller,  S.\",\"Font,  J.\",\"Liew,  C.\",\"Cram,  E.\",\"Schroeder,  T.\",\"Turner,  J.\",\"Crossley,  M.\",\"Mackay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wissmueller-font-liew-cram-schroeder-turner-crossley-mackay-etal-proteinproteininteractionsanalysisofafalsepositivegstpulldownresult-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Structural basis and specificity of acetylated transcription factor GATA1 recognition by BET family bromodomain protein Brd3\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"2632-2640\",\"volume\":\"31\",\"id\":\"429de600-d97d-3c2c-9f33-97c9b5577dcd\",\"created\":\"2023-01-10T01:45:09.655Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:09.655Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Recent data demonstrate that small synthetic compounds specifically targeting bromodomain proteins can modulate the expression of cancer-related or inflammatory genes. Although these studies have focused on the ability of bromodomains to recognize acetylated histones, it is increasingly becoming clear that histone-like modifications exist on other important proteins, such as transcription factors. However, our understanding of the molecular mechanisms through which these modifications modulate protein function is far from complete. The transcription factor GATA1 can be acetylated at lysine residues adjacent to the zinc finger domains, and this acetylation is essential for the normal chromatin occupancy of GATA1. We have recently identified the bromodomain-containing protein Brd3 as a cofactor that interacts with acetylated GATA1 and shown that this interaction is essential for the targeting of GATA1 to chromatin. Here we describe the structural basis for this interaction. Our data reveal for the first time the molecular details of an interaction between a transcription factor bearing multiple acetylation modifications and its cognate recognition module. We also show that this interaction can be inhibited by an acetyllysine mimic, highlighting the importance of further increasing the specificity of compounds that target bromodomain and extraterminal (BET) bromodomains in order to fully realize their therapeutic potential. © 2011, American Society for Microbiology.\",\"bibtype\":\"article\",\"author\":\"Gamsjaeger, R. and Webb, S.R. and Lamonica, J.M. and Billin, A. and Blobel, G.A. and Mackay, J.P.\",\"doi\":\"10.1128/MCB.05413-11\",\"journal\":\"Molecular and Cellular Biology\",\"number\":\"13\",\"bibtex\":\"@article{\\n title = {Structural basis and specificity of acetylated transcription factor GATA1 recognition by BET family bromodomain protein Brd3},\\n type = {article},\\n year = {2011},\\n pages = {2632-2640},\\n volume = {31},\\n id = {429de600-d97d-3c2c-9f33-97c9b5577dcd},\\n created = {2023-01-10T01:45:09.655Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:09.655Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Recent data demonstrate that small synthetic compounds specifically targeting bromodomain proteins can modulate the expression of cancer-related or inflammatory genes. Although these studies have focused on the ability of bromodomains to recognize acetylated histones, it is increasingly becoming clear that histone-like modifications exist on other important proteins, such as transcription factors. However, our understanding of the molecular mechanisms through which these modifications modulate protein function is far from complete. The transcription factor GATA1 can be acetylated at lysine residues adjacent to the zinc finger domains, and this acetylation is essential for the normal chromatin occupancy of GATA1. We have recently identified the bromodomain-containing protein Brd3 as a cofactor that interacts with acetylated GATA1 and shown that this interaction is essential for the targeting of GATA1 to chromatin. Here we describe the structural basis for this interaction. Our data reveal for the first time the molecular details of an interaction between a transcription factor bearing multiple acetylation modifications and its cognate recognition module. We also show that this interaction can be inhibited by an acetyllysine mimic, highlighting the importance of further increasing the specificity of compounds that target bromodomain and extraterminal (BET) bromodomains in order to fully realize their therapeutic potential. © 2011, American Society for Microbiology.},\\n bibtype = {article},\\n author = {Gamsjaeger, R. and Webb, S.R. and Lamonica, J.M. and Billin, A. and Blobel, G.A. and Mackay, J.P.},\\n doi = {10.1128/MCB.05413-11},\\n journal = {Molecular and Cellular Biology},\\n number = {13}\\n}\",\"author_short\":[\"Gamsjaeger,  R.\",\"Webb,  S.\",\"Lamonica,  J.\",\"Billin,  A.\",\"Blobel,  G.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gamsjaeger-webb-lamonica-billin-blobel-mackay-structuralbasisandspecificityofacetylatedtranscriptionfactorgata1recognitionbybetfamilybromodomainproteinbrd3-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Plant homeodomain (PHD) fingers of CHD4 are histone H3-binding modules with preference for unmodified H3K4 and methylated H3K9\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"11779-11791\",\"volume\":\"286\",\"id\":\"5cbc3d31-ec2f-35ec-a0c2-85fb87abd270\",\"created\":\"2023-01-10T01:45:10.576Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:10.576Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"A major challenge in chromatin biology is to understand the mechanisms by which chromatin is remodeled into active or inactive states as required during development and cell differentiation. One complex implicated in these processes is the nucleosome remodeling and histone deacetylase (NuRD) complex, which contains both histone deacetylase and nucleosome remodeling activities and has been implicated in the silencing of subsets of genes involved in various stages of cellular development. Chromodomain-helicase-DNA-binding protein 4 (CHD4) is a core component of the NuRD complex and contains a nucleosome remodeling ATPase domain along with two chromodomains and two plant homeodomain (PHD) fingers. We have previously demonstrated that the second PHD finger of CHD4 binds peptides corresponding to the N terminus of histone H3 methylated at Lys9. Here, we determine the solution structure of PHD2 in complex with H3K9me3, revealing the molecular basis of histone recognition, including a cation-π recognition mechanism for methylated Lys9. Additionally, we demonstrate that the first PHD finger also exhibits binding to the N terminus of H3, and we establish the histone-binding surface of this domain. This is the first instance where histone binding ability has been demonstrated for two separate PHD modules within the one protein. These findings suggest that CHD4 could bind to two H3 N-terminal tails on the same nucleosome or on two separate nucleosomes simultaneously, presenting exciting implications for the mechanism by which CHD4 and the NuRD complex could direct chromatin remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Mansfield, R.E. and Musselman, C.A. and Kwan, A.H. and Oliver, S.S. and Garske, A.L. and Davrazou, F. and Denu, J.M. and Kutateladze, T.G. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M110.208207\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"13\",\"bibtex\":\"@article{\\n title = {Plant homeodomain (PHD) fingers of CHD4 are histone H3-binding modules with preference for unmodified H3K4 and methylated H3K9},\\n type = {article},\\n year = {2011},\\n pages = {11779-11791},\\n volume = {286},\\n id = {5cbc3d31-ec2f-35ec-a0c2-85fb87abd270},\\n created = {2023-01-10T01:45:10.576Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:10.576Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {A major challenge in chromatin biology is to understand the mechanisms by which chromatin is remodeled into active or inactive states as required during development and cell differentiation. One complex implicated in these processes is the nucleosome remodeling and histone deacetylase (NuRD) complex, which contains both histone deacetylase and nucleosome remodeling activities and has been implicated in the silencing of subsets of genes involved in various stages of cellular development. Chromodomain-helicase-DNA-binding protein 4 (CHD4) is a core component of the NuRD complex and contains a nucleosome remodeling ATPase domain along with two chromodomains and two plant homeodomain (PHD) fingers. We have previously demonstrated that the second PHD finger of CHD4 binds peptides corresponding to the N terminus of histone H3 methylated at Lys9. Here, we determine the solution structure of PHD2 in complex with H3K9me3, revealing the molecular basis of histone recognition, including a cation-π recognition mechanism for methylated Lys9. Additionally, we demonstrate that the first PHD finger also exhibits binding to the N terminus of H3, and we establish the histone-binding surface of this domain. This is the first instance where histone binding ability has been demonstrated for two separate PHD modules within the one protein. These findings suggest that CHD4 could bind to two H3 N-terminal tails on the same nucleosome or on two separate nucleosomes simultaneously, presenting exciting implications for the mechanism by which CHD4 and the NuRD complex could direct chromatin remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Mansfield, R.E. and Musselman, C.A. and Kwan, A.H. and Oliver, S.S. and Garske, A.L. and Davrazou, F. and Denu, J.M. and Kutateladze, T.G. and Mackay, J.P.},\\n doi = {10.1074/jbc.M110.208207},\\n journal = {Journal of Biological Chemistry},\\n number = {13}\\n}\",\"author_short\":[\"Mansfield,  R.\",\"Musselman,  C.\",\"Kwan,  A.\",\"Oliver,  S.\",\"Garske,  A.\",\"Davrazou,  F.\",\"Denu,  J.\",\"Kutateladze,  T.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mansfield-musselman-kwan-oliver-garske-davrazou-denu-kutateladze-etal-planthomeodomainphdfingersofchd4arehistoneh3bindingmoduleswithpreferenceforunmodifiedh3k4andmethylatedh3k9-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Characterization of a family of RanBP2-Type zinc fingers that can recognize single-stranded RNA\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"273-283\",\"volume\":\"407\",\"id\":\"48de412f-0ebf-3005-9f78-2ff45c65410e\",\"created\":\"2023-01-10T01:45:11.498Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:11.498Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The recognition of single-stranded RNA (ssRNA) is an important aspect of gene regulation, and a number of different classes of protein domains that recognize ssRNA in a sequence-specific manner have been identified. Recently, we demonstrated that the RanBP2-type zinc finger (ZnF) domains from the human splicing factor ZnF Ran binding domain-containing protein 2 (ZRANB2) can bind to a sequence containing the consensus AGGUAA. Six other human proteins, namely, Ewing's sarcoma (EWS), translocated in liposarcoma (TLS)/FUS, RNA-binding protein 56 (RBP56), RNA-binding motif 5 (RBM5), RNA-binding motif 10 (RBM10) and testis-expressed sequence 13A (TEX13A), each contains a single ZnF with homology to the ZRANB2 ZnFs, and several of these proteins have been implicated in the regulation of mRNA processing. Here, we show that all of these ZnFs are able to bind with micromolar affinities to ssRNA containing a GGU motif. NMR titration data reveal that binding is mediated by the corresponding surfaces on each ZnF, and we also show that sequence selectivity is largely limited to the GGU core motif and that substitution of the three flanking adenines that were selected in our original selection experiment has a minimal effect on binding affinity. These data establish a subset of RanBP2-type ZnFs as a new family of ssRNA-binding motifs. © 2011 Elsevier Ltd.\",\"bibtype\":\"article\",\"author\":\"Nguyen, C.D. and Mansfield, R.E. and Leung, W. and Vaz, P.M. and Loughlin, F.E. and Grant, R.P. and MacKay, J.P.\",\"doi\":\"10.1016/j.jmb.2010.12.041\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Characterization of a family of RanBP2-Type zinc fingers that can recognize single-stranded RNA},\\n type = {article},\\n year = {2011},\\n pages = {273-283},\\n volume = {407},\\n id = {48de412f-0ebf-3005-9f78-2ff45c65410e},\\n created = {2023-01-10T01:45:11.498Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:11.498Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The recognition of single-stranded RNA (ssRNA) is an important aspect of gene regulation, and a number of different classes of protein domains that recognize ssRNA in a sequence-specific manner have been identified. Recently, we demonstrated that the RanBP2-type zinc finger (ZnF) domains from the human splicing factor ZnF Ran binding domain-containing protein 2 (ZRANB2) can bind to a sequence containing the consensus AGGUAA. Six other human proteins, namely, Ewing's sarcoma (EWS), translocated in liposarcoma (TLS)/FUS, RNA-binding protein 56 (RBP56), RNA-binding motif 5 (RBM5), RNA-binding motif 10 (RBM10) and testis-expressed sequence 13A (TEX13A), each contains a single ZnF with homology to the ZRANB2 ZnFs, and several of these proteins have been implicated in the regulation of mRNA processing. Here, we show that all of these ZnFs are able to bind with micromolar affinities to ssRNA containing a GGU motif. NMR titration data reveal that binding is mediated by the corresponding surfaces on each ZnF, and we also show that sequence selectivity is largely limited to the GGU core motif and that substitution of the three flanking adenines that were selected in our original selection experiment has a minimal effect on binding affinity. These data establish a subset of RanBP2-type ZnFs as a new family of ssRNA-binding motifs. © 2011 Elsevier Ltd.},\\n bibtype = {article},\\n author = {Nguyen, C.D. and Mansfield, R.E. and Leung, W. and Vaz, P.M. and Loughlin, F.E. and Grant, R.P. and MacKay, J.P.},\\n doi = {10.1016/j.jmb.2010.12.041},\\n journal = {Journal of Molecular Biology},\\n number = {2}\\n}\",\"author_short\":[\"Nguyen,  C.\",\"Mansfield,  R.\",\"Leung,  W.\",\"Vaz,  P.\",\"Loughlin,  F.\",\"Grant,  R.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"nguyen-mansfield-leung-vaz-loughlin-grant-mackay-characterizationofafamilyofranbp2typezincfingersthatcanrecognizesinglestrandedrna-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"14443-14448\",\"volume\":\"108\",\"id\":\"765574bf-c2e8-3a13-8a8c-db663be1661f\",\"created\":\"2023-01-10T01:45:12.413Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:12.413Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (<50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.\",\"bibtype\":\"article\",\"author\":\"Wilkinson-White, L. and Gamsjaeger, R. and Dastmalchi, S. and Wienert, B. and Stokes, P.H. and Crossley, M. and Mackay, J.P. and Matthews, J.M.\",\"doi\":\"10.1073/pnas.1105898108\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"35\",\"bibtex\":\"@article{\\n title = {Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1},\\n type = {article},\\n year = {2011},\\n pages = {14443-14448},\\n volume = {108},\\n id = {765574bf-c2e8-3a13-8a8c-db663be1661f},\\n created = {2023-01-10T01:45:12.413Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:12.413Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (<50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.},\\n bibtype = {article},\\n author = {Wilkinson-White, L. and Gamsjaeger, R. and Dastmalchi, S. and Wienert, B. and Stokes, P.H. and Crossley, M. and Mackay, J.P. and Matthews, J.M.},\\n doi = {10.1073/pnas.1105898108},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {35}\\n}\",\"author_short\":[\"Wilkinson-White,  L.\",\"Gamsjaeger,  R.\",\"Dastmalchi,  S.\",\"Wienert,  B.\",\"Stokes,  P.\",\"Crossley,  M.\",\"Mackay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Macromolecular NMR spectroscopy for the non-spectroscopist: Beyond macromolecular solution structure determination\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"704-715\",\"volume\":\"278\",\"id\":\"0c69b0a8-cbe5-3ad1-acb3-27b8003b448f\",\"created\":\"2023-01-10T01:45:13.329Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:13.329Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"A strength of NMR spectroscopy is its ability to monitor, on an atomic level, molecular changes and interactions. In this review, which is intended for non-spectroscopist, we describe major uses of NMR in protein science beyond solution structure determination. After first touching on how NMR can be used to quickly determine whether a mutation induces structural perturbations in a protein, we describe the unparalleled ability of NMR to monitor binding interactions over a wide range of affinities, molecular masses and solution conditions. We discuss the use of NMR to measure the dynamics of proteins at the atomic level and over a wide range of timescales. Finally, we outline new and expanding areas such as macromolecular structure determination in multicomponent systems, as well as in the solid state and in vivo. © 2011 FEBS.\",\"bibtype\":\"article\",\"author\":\"Bieri, M. and Kwan, A.H. and Mobli, M. and King, G.F. and MacKay, J.P. and Gooley, P.R.\",\"doi\":\"10.1111/j.1742-4658.2011.08005.x\",\"journal\":\"FEBS Journal\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Macromolecular NMR spectroscopy for the non-spectroscopist: Beyond macromolecular solution structure determination},\\n type = {article},\\n year = {2011},\\n pages = {704-715},\\n volume = {278},\\n id = {0c69b0a8-cbe5-3ad1-acb3-27b8003b448f},\\n created = {2023-01-10T01:45:13.329Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:13.329Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {A strength of NMR spectroscopy is its ability to monitor, on an atomic level, molecular changes and interactions. In this review, which is intended for non-spectroscopist, we describe major uses of NMR in protein science beyond solution structure determination. After first touching on how NMR can be used to quickly determine whether a mutation induces structural perturbations in a protein, we describe the unparalleled ability of NMR to monitor binding interactions over a wide range of affinities, molecular masses and solution conditions. We discuss the use of NMR to measure the dynamics of proteins at the atomic level and over a wide range of timescales. Finally, we outline new and expanding areas such as macromolecular structure determination in multicomponent systems, as well as in the solid state and in vivo. © 2011 FEBS.},\\n bibtype = {article},\\n author = {Bieri, M. and Kwan, A.H. and Mobli, M. and King, G.F. and MacKay, J.P. and Gooley, P.R.},\\n doi = {10.1111/j.1742-4658.2011.08005.x},\\n journal = {FEBS Journal},\\n number = {5}\\n}\",\"author_short\":[\"Bieri,  M.\",\"Kwan,  A.\",\"Mobli,  M.\",\"King,  G.\",\"MacKay,  J.\",\"Gooley,  P.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"bieri-kwan-mobli-king-mackay-gooley-macromolecularnmrspectroscopyforthenonspectroscopistbeyondmacromolecularsolutionstructuredetermination-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Erratum: The prospects for designer single-stranded RNA-binding proteins (Nature Structural and Molecular Biology (2011) 18 (256-261))\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"516\",\"volume\":\"18\",\"id\":\"af45e7f3-95df-3e38-b1e4-265877fed464\",\"created\":\"2023-01-10T01:45:14.370Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:14.370Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"MacKay, J.P. and Font, J. and Segal, D.J.\",\"doi\":\"10.1038/nsmb0411-516e\",\"journal\":\"Nature Structural and Molecular Biology\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {Erratum: The prospects for designer single-stranded RNA-binding proteins (Nature Structural and Molecular Biology (2011) 18 (256-261))},\\n type = {article},\\n year = {2011},\\n pages = {516},\\n volume = {18},\\n id = {af45e7f3-95df-3e38-b1e4-265877fed464},\\n created = {2023-01-10T01:45:14.370Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:14.370Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {MacKay, J.P. and Font, J. and Segal, D.J.},\\n doi = {10.1038/nsmb0411-516e},\\n journal = {Nature Structural and Molecular Biology},\\n number = {4}\\n}\",\"author_short\":[\"MacKay,  J.\",\"Font,  J.\",\"Segal,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-font-segal-erratumtheprospectsfordesignersinglestrandedrnabindingproteinsnaturestructuralandmolecularbiology201118256261-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Macromolecular NMR spectroscopy for the non-spectroscopist\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"687-703\",\"volume\":\"278\",\"id\":\"ed62160d-f565-3f31-ab39-6f55529dd09e\",\"created\":\"2023-01-10T01:45:15.274Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:15.274Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"NMR spectroscopy is a powerful tool for studying the structure, function and dynamics of biological macromolecules. However, non-spectroscopists often find NMR theory daunting and data interpretation nontrivial. As the first of two back-to-back reviews on NMR spectroscopy aimed at non-spectroscopists, the present review first provides an introduction to the basics of macromolecular NMR spectroscopy, including a discussion of typical sample requirements and what information can be obtained from simple NMR experiments. We then review the use of NMR spectroscopy for determining the 3D structures of macromolecules and examine how to judge the quality of NMR-derived structures. © 2011 FEBS.\",\"bibtype\":\"article\",\"author\":\"Kwan, A.H. and Mobli, M. and Gooley, P.R. and King, G.F. and MacKay, J.P.\",\"doi\":\"10.1111/j.1742-4658.2011.08004.x\",\"journal\":\"FEBS Journal\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Macromolecular NMR spectroscopy for the non-spectroscopist},\\n type = {article},\\n year = {2011},\\n pages = {687-703},\\n volume = {278},\\n id = {ed62160d-f565-3f31-ab39-6f55529dd09e},\\n created = {2023-01-10T01:45:15.274Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:15.274Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {NMR spectroscopy is a powerful tool for studying the structure, function and dynamics of biological macromolecules. However, non-spectroscopists often find NMR theory daunting and data interpretation nontrivial. As the first of two back-to-back reviews on NMR spectroscopy aimed at non-spectroscopists, the present review first provides an introduction to the basics of macromolecular NMR spectroscopy, including a discussion of typical sample requirements and what information can be obtained from simple NMR experiments. We then review the use of NMR spectroscopy for determining the 3D structures of macromolecules and examine how to judge the quality of NMR-derived structures. © 2011 FEBS.},\\n bibtype = {article},\\n author = {Kwan, A.H. and Mobli, M. and Gooley, P.R. and King, G.F. and MacKay, J.P.},\\n doi = {10.1111/j.1742-4658.2011.08004.x},\\n journal = {FEBS Journal},\\n number = {5}\\n}\",\"author_short\":[\"Kwan,  A.\",\"Mobli,  M.\",\"Gooley,  P.\",\"King,  G.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kwan-mobli-gooley-king-mackay-macromolecularnmrspectroscopyforthenonspectroscopist-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Insights into association of the NuRD complex with FOG-1 from the crystal structure of an RbAp48·FOG-1 complex\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"1196-1203\",\"volume\":\"286\",\"id\":\"0dcda68f-44aa-33d1-afea-ef433768b02c\",\"created\":\"2023-01-10T01:45:16.197Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:16.197Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Chromatin-modifying complexes such as the NuRD complex are recruited to particular genomic sites by gene-specific nuclear factors. Overall, however, little is known about the molecular basis for these interactions. Here, we present the 1.9 Å resolution crystal structure of the NuRD subunit RbAp48 bound to the 15 N-terminal amino acids of the GATA-1 cofactor FOG-1. The FOG-1 peptide contacts a negatively charged binding pocket on top of the RbAp48 β-propeller that is distinct from the binding surface used by RpAp48 to contact histone H4. We further show that RbAp48 interacts with the NuRD subunit MTA-1 via a surface that is distinct from its FOG-binding pocket, providing a first glimpse into the way in which NuRD assembly facilitates interactions with cofactors. Our RbAp48·FOG-1 structure provides insight into the molecular determinants of FOG-1-dependent association with the NuRD complex and into the links between transcription regulation and nucleosome remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Lejon, S. and Thong, S.Y. and Murthy, A. and AlQarni, S. and Murzina, N.V. and Blobel, G.A. and Laue, E.D. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M110.195842\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Insights into association of the NuRD complex with FOG-1 from the crystal structure of an RbAp48·FOG-1 complex},\\n type = {article},\\n year = {2011},\\n pages = {1196-1203},\\n volume = {286},\\n id = {0dcda68f-44aa-33d1-afea-ef433768b02c},\\n created = {2023-01-10T01:45:16.197Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:16.197Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Chromatin-modifying complexes such as the NuRD complex are recruited to particular genomic sites by gene-specific nuclear factors. Overall, however, little is known about the molecular basis for these interactions. Here, we present the 1.9 Å resolution crystal structure of the NuRD subunit RbAp48 bound to the 15 N-terminal amino acids of the GATA-1 cofactor FOG-1. The FOG-1 peptide contacts a negatively charged binding pocket on top of the RbAp48 β-propeller that is distinct from the binding surface used by RpAp48 to contact histone H4. We further show that RbAp48 interacts with the NuRD subunit MTA-1 via a surface that is distinct from its FOG-binding pocket, providing a first glimpse into the way in which NuRD assembly facilitates interactions with cofactors. Our RbAp48·FOG-1 structure provides insight into the molecular determinants of FOG-1-dependent association with the NuRD complex and into the links between transcription regulation and nucleosome remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Lejon, S. and Thong, S.Y. and Murthy, A. and AlQarni, S. and Murzina, N.V. and Blobel, G.A. and Laue, E.D. and Mackay, J.P.},\\n doi = {10.1074/jbc.M110.195842},\\n journal = {Journal of Biological Chemistry},\\n number = {2}\\n}\",\"author_short\":[\"Lejon,  S.\",\"Thong,  S.\",\"Murthy,  A.\",\"AlQarni,  S.\",\"Murzina,  N.\",\"Blobel,  G.\",\"Laue,  E.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"lejon-thong-murthy-alqarni-murzina-blobel-laue-mackay-insightsintoassociationofthenurdcomplexwithfog1fromthecrystalstructureofanrbap48fog1complex-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The prospects for designer single-stranded RNA-binding proteins\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"256-261\",\"volume\":\"18\",\"id\":\"ff45d397-50a9-36fc-8a33-31782437940d\",\"created\":\"2023-01-10T01:45:17.102Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:17.102Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Spectacular progress has been made in the design of proteins that recognize double-stranded DNA with a chosen specificity, to the point that designer DNA-binding proteins can be ordered commercially. This success raises the question of whether it will be possible to engineer libraries of proteins that can recognize RNA with tailored specificity. Given the recent explosion in the number and diversity of RNA species demonstrated to play roles in biology, designer RNA-binding proteins are set to become valuable tools, both in the research laboratory and potentially in the clinic. Here we discuss the prospects for the realization of this idea. © 2011 Nature America, Inc. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"MacKay, J.P. and Font, J. and Segal, D.J.\",\"doi\":\"10.1038/nsmb.2005\",\"journal\":\"Nature Structural and Molecular Biology\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {The prospects for designer single-stranded RNA-binding proteins},\\n type = {article},\\n year = {2011},\\n pages = {256-261},\\n volume = {18},\\n id = {ff45d397-50a9-36fc-8a33-31782437940d},\\n created = {2023-01-10T01:45:17.102Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:17.102Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Spectacular progress has been made in the design of proteins that recognize double-stranded DNA with a chosen specificity, to the point that designer DNA-binding proteins can be ordered commercially. This success raises the question of whether it will be possible to engineer libraries of proteins that can recognize RNA with tailored specificity. Given the recent explosion in the number and diversity of RNA species demonstrated to play roles in biology, designer RNA-binding proteins are set to become valuable tools, both in the research laboratory and potentially in the clinic. Here we discuss the prospects for the realization of this idea. © 2011 Nature America, Inc. All rights reserved.},\\n bibtype = {article},\\n author = {MacKay, J.P. and Font, J. and Segal, D.J.},\\n doi = {10.1038/nsmb.2005},\\n journal = {Nature Structural and Molecular Biology},\\n number = {3}\\n}\",\"author_short\":[\"MacKay,  J.\",\"Font,  J.\",\"Segal,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-font-segal-theprospectsfordesignersinglestrandedrnabindingproteins-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The structure of a truncated phosphoribosylanthranilate isomerase suggests a unified model for evolution of the (βα)<inf>8</inf> barrel fold\",\"type\":\"article\",\"year\":\"2011\",\"pages\":\"291-303\",\"volume\":\"408\",\"id\":\"484786a4-2a38-33ce-8896-5cba612e7ef8\",\"created\":\"2023-01-10T01:45:18.013Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:18.013Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The (βα)8 barrel is one of the most common protein folds, and enzymes with this architecture display a remarkable range of catalytic activities. Many of these functions are associated with ancient metabolic pathways, and phylogenetic reconstructions suggest that the (βα)8 barrel was one of the very first protein folds to emerge. Consequently, there is considerable interest in understanding the evolutionary processes that gave rise to this fold. In particular, much attention has been focused on the plausibility of (βα)8 barrel evolution from homodimers of half barrels. However, we previously isolated a three-quarter-barrel-sized fragment of a (βα)8 barrel, termed truncated phosphoribosylanthranilate isomerase (trPRAI), that is soluble and almost as thermostable as full-length N-(5′-phosphoribosyl) anthranilate isomerase (PRAI). Here, we report the NMR-derived structure of trPRAI. The subdomain is monomeric, is well ordered and adopts a native-like structure in solution. Side chains from strands β1 (Glu3 and Lys5), β2 (Tyr25) and β6 (Lys122) of trPRAI repack to shield the hydrophobic core from the solvent. This result demonstrates that three-quarter barrels were viable intermediates in the evolution of the (βα)8 barrel fold. We propose a unified model for (βα)8 barrel evolution that combines our data, previously published work and plausible scenarios for the emergence of (initially error-prone) genetic systems. In this model, the earliest proto-cells contained diverse pools of part-barrel subdomains. Combinatorial assembly of these subdomains gave rise to many distinct lineages of (βα)8 barrel proteins, that is, our model excludes the possibility that there was a single (βα)8 barrel from which all present examples are descended. © 2011 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Setiyaputra, S. and MacKay, J.P. and Patrick, W.M.\",\"doi\":\"10.1016/j.jmb.2011.02.048\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {The structure of a truncated phosphoribosylanthranilate isomerase suggests a unified model for evolution of the (βα)<inf>8</inf> barrel fold},\\n type = {article},\\n year = {2011},\\n pages = {291-303},\\n volume = {408},\\n id = {484786a4-2a38-33ce-8896-5cba612e7ef8},\\n created = {2023-01-10T01:45:18.013Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:18.013Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The (βα)8 barrel is one of the most common protein folds, and enzymes with this architecture display a remarkable range of catalytic activities. Many of these functions are associated with ancient metabolic pathways, and phylogenetic reconstructions suggest that the (βα)8 barrel was one of the very first protein folds to emerge. Consequently, there is considerable interest in understanding the evolutionary processes that gave rise to this fold. In particular, much attention has been focused on the plausibility of (βα)8 barrel evolution from homodimers of half barrels. However, we previously isolated a three-quarter-barrel-sized fragment of a (βα)8 barrel, termed truncated phosphoribosylanthranilate isomerase (trPRAI), that is soluble and almost as thermostable as full-length N-(5′-phosphoribosyl) anthranilate isomerase (PRAI). Here, we report the NMR-derived structure of trPRAI. The subdomain is monomeric, is well ordered and adopts a native-like structure in solution. Side chains from strands β1 (Glu3 and Lys5), β2 (Tyr25) and β6 (Lys122) of trPRAI repack to shield the hydrophobic core from the solvent. This result demonstrates that three-quarter barrels were viable intermediates in the evolution of the (βα)8 barrel fold. We propose a unified model for (βα)8 barrel evolution that combines our data, previously published work and plausible scenarios for the emergence of (initially error-prone) genetic systems. In this model, the earliest proto-cells contained diverse pools of part-barrel subdomains. Combinatorial assembly of these subdomains gave rise to many distinct lineages of (βα)8 barrel proteins, that is, our model excludes the possibility that there was a single (βα)8 barrel from which all present examples are descended. © 2011 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Setiyaputra, S. and MacKay, J.P. and Patrick, W.M.},\\n doi = {10.1016/j.jmb.2011.02.048},\\n journal = {Journal of Molecular Biology},\\n number = {2}\\n}\",\"author_short\":[\"Setiyaputra,  S.\",\"MacKay,  J.\",\"Patrick,  W.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"setiyaputra-mackay-patrick-thestructureofatruncatedphosphoribosylanthranilateisomerasesuggestsaunifiedmodelforevolutionoftheinf8infbarrelfold-2011\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"AHSP (α-haemoglobin-stabilizing protein) stabilizes apo-α-haemoglobin in a partially folded state\",\"type\":\"article\",\"year\":\"2010\",\"pages\":\"275-282\",\"volume\":\"432\",\"id\":\"cfeb53fe-7617-3568-b0ac-944abaedb834\",\"created\":\"2023-01-10T01:45:18.928Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:18.928Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"To produce functional Hb (haemoglobin), nascent α-globin (αo) and β-globin (βo) chains must each bind a single haem molecule (to form αh and βh) and interact together to form heterodimers.The precise sequence of binding events is unknown, and it has been suggested that additional factors might enhance the efficiency of Hb folding. AHSP (α-haemoglobin-stabilizing protein) has been shown previously to bind αh and regulate redox activity of the haem iron. In the present study, we used a combination of classical and dynamic light scattering and NMR spectroscopy to demonstrate that AHSP forms a heterodimeric complex with αo that inhibits αo aggregation and promotes αo folding in the absence of haem. These findings indicate that AHSP may function as an αo-specific chaperone, and suggest an important role for αo in guiding Hb assembly by stabilizing βo and inhibiting off-pathway self-association of βh. © The Authors Journal compilation © 2010 Biochemical Society.\",\"bibtype\":\"article\",\"author\":\"Krishna Kumar, K. and Dickson, C.F. and Weiss, M.J. and Mackay, J.P. and Gell, D.A.\",\"doi\":\"10.1042/BJ20100642\",\"journal\":\"Biochemical Journal\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {AHSP (α-haemoglobin-stabilizing protein) stabilizes apo-α-haemoglobin in a partially folded state},\\n type = {article},\\n year = {2010},\\n pages = {275-282},\\n volume = {432},\\n id = {cfeb53fe-7617-3568-b0ac-944abaedb834},\\n created = {2023-01-10T01:45:18.928Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:18.928Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {To produce functional Hb (haemoglobin), nascent α-globin (αo) and β-globin (βo) chains must each bind a single haem molecule (to form αh and βh) and interact together to form heterodimers.The precise sequence of binding events is unknown, and it has been suggested that additional factors might enhance the efficiency of Hb folding. AHSP (α-haemoglobin-stabilizing protein) has been shown previously to bind αh and regulate redox activity of the haem iron. In the present study, we used a combination of classical and dynamic light scattering and NMR spectroscopy to demonstrate that AHSP forms a heterodimeric complex with αo that inhibits αo aggregation and promotes αo folding in the absence of haem. These findings indicate that AHSP may function as an αo-specific chaperone, and suggest an important role for αo in guiding Hb assembly by stabilizing βo and inhibiting off-pathway self-association of βh. © The Authors Journal compilation © 2010 Biochemical Society.},\\n bibtype = {article},\\n author = {Krishna Kumar, K. and Dickson, C.F. and Weiss, M.J. and Mackay, J.P. and Gell, D.A.},\\n doi = {10.1042/BJ20100642},\\n journal = {Biochemical Journal},\\n number = {2}\\n}\",\"author_short\":[\"Krishna Kumar,  K.\",\"Dickson,  C.\",\"Weiss,  M.\",\"Mackay,  J.\",\"Gell,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"krishnakumar-dickson-weiss-mackay-gell-ahsphaemoglobinstabilizingproteinstabilizesapohaemoglobininapartiallyfoldedstate-2010\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Beyond DNA: Zinc finger domains as RNA-binding modules\",\"type\":\"book\",\"year\":\"2010\",\"source\":\"Methods in Molecular Biology\",\"pages\":\"479-491\",\"volume\":\"649\",\"id\":\"c6c74829-cc59-3c5a-bd21-95b1c21c0aac\",\"created\":\"2023-01-10T01:45:19.842Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:19.842Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.\",\"bibtype\":\"book\",\"author\":\"Font, J. and MacKay, J.P.\",\"doi\":\"10.1007/978-1-60761-753-2_29\",\"bibtex\":\"@book{\\n title = {Beyond DNA: Zinc finger domains as RNA-binding modules},\\n type = {book},\\n year = {2010},\\n source = {Methods in Molecular Biology},\\n pages = {479-491},\\n volume = {649},\\n id = {c6c74829-cc59-3c5a-bd21-95b1c21c0aac},\\n created = {2023-01-10T01:45:19.842Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:19.842Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.},\\n bibtype = {book},\\n author = {Font, J. and MacKay, J.P.},\\n doi = {10.1007/978-1-60761-753-2_29}\\n}\",\"author_short\":[\"Font,  J.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"font-mackay-beyonddnazincfingerdomainsasrnabindingmodules-2010\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Preface\",\"type\":\"book\",\"year\":\"2010\",\"source\":\"Methods in Molecular Biology\",\"volume\":\"649\",\"id\":\"7ce2996b-3608-389e-b725-b249c989c5b4\",\"created\":\"2023-01-10T01:45:20.744Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:20.744Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"book\",\"author\":\"MacKay, J.P. and Segal, D.J.\",\"bibtex\":\"@book{\\n title = {Preface},\\n type = {book},\\n year = {2010},\\n source = {Methods in Molecular Biology},\\n volume = {649},\\n id = {7ce2996b-3608-389e-b725-b249c989c5b4},\\n created = {2023-01-10T01:45:20.744Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:20.744Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {book},\\n author = {MacKay, J.P. and Segal, D.J.}\\n}\",\"author_short\":[\"MacKay,  J.\",\"Segal,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-segal-preface-2010\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular Lmo2-LIM2/Ldb1-LID complex\",\"type\":\"article\",\"year\":\"2010\",\"pages\":\"203-206\",\"volume\":\"4\",\"id\":\"d3067811-3bab-3f57-99ac-522a48b4c9b3\",\"created\":\"2023-01-10T01:45:21.656Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:21.656Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Lmo2 is a LIM-only protein involved in hematopoiesis and the development of T-cell acute lymphoblastic leukaemia. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the C-terminal LIM domain from Lmo2 tethered to the LIM interaction domain (LID) from LIM domain binding protein 1 (Ldb1). © 2010 Springer Science+Business Media B.V.\",\"bibtype\":\"article\",\"author\":\"Wilkinson-White, L.E. and Dastmalchi, S. and Kwan, A.H. and Ryan, D.P. and MacKay, J.P. and Matthews, J.M.\",\"doi\":\"10.1007/s12104-010-9240-y\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular Lmo2-LIM2/Ldb1-LID complex},\\n type = {article},\\n year = {2010},\\n pages = {203-206},\\n volume = {4},\\n id = {d3067811-3bab-3f57-99ac-522a48b4c9b3},\\n created = {2023-01-10T01:45:21.656Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:21.656Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Lmo2 is a LIM-only protein involved in hematopoiesis and the development of T-cell acute lymphoblastic leukaemia. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the C-terminal LIM domain from Lmo2 tethered to the LIM interaction domain (LID) from LIM domain binding protein 1 (Ldb1). © 2010 Springer Science+Business Media B.V.},\\n bibtype = {article},\\n author = {Wilkinson-White, L.E. and Dastmalchi, S. and Kwan, A.H. and Ryan, D.P. and MacKay, J.P. and Matthews, J.M.},\\n doi = {10.1007/s12104-010-9240-y},\\n journal = {Biomolecular NMR Assignments},\\n number = {2}\\n}\",\"author_short\":[\"Wilkinson-White,  L.\",\"Dastmalchi,  S.\",\"Kwan,  A.\",\"Ryan,  D.\",\"MacKay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wilkinsonwhite-dastmalchi-kwan-ryan-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo2lim2ldb1lidcomplex-2010\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Two-state conformational equilibrium in the Par-4 leucine zipper domain\",\"type\":\"article\",\"year\":\"2010\",\"pages\":\"2433-2449\",\"volume\":\"78\",\"id\":\"f7152d3f-03a8-3aab-8417-8bbf484d2488\",\"created\":\"2023-01-10T01:45:22.571Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:22.571Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.\",\"bibtype\":\"article\",\"author\":\"Schwalbe, M. and Dutta, K. and Libich, D.S. and Venugopal, H. and Claridge, J.K. and Gell, D.A. and Mackay, J.P. and Edwards, P.J.B. and Pascal, S.M.\",\"doi\":\"10.1002/prot.22752\",\"journal\":\"Proteins: Structure, Function and Bioinformatics\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Two-state conformational equilibrium in the Par-4 leucine zipper domain},\\n type = {article},\\n year = {2010},\\n pages = {2433-2449},\\n volume = {78},\\n id = {f7152d3f-03a8-3aab-8417-8bbf484d2488},\\n created = {2023-01-10T01:45:22.571Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:22.571Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.},\\n bibtype = {article},\\n author = {Schwalbe, M. and Dutta, K. and Libich, D.S. and Venugopal, H. and Claridge, J.K. and Gell, D.A. and Mackay, J.P. and Edwards, P.J.B. and Pascal, S.M.},\\n doi = {10.1002/prot.22752},\\n journal = {Proteins: Structure, Function and Bioinformatics},\\n number = {11}\\n}\",\"author_short\":[\"Schwalbe,  M.\",\"Dutta,  K.\",\"Libich,  D.\",\"Venugopal,  H.\",\"Claridge,  J.\",\"Gell,  D.\",\"Mackay,  J.\",\"Edwards,  P.\",\"Pascal,  S.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis\",\"type\":\"article\",\"year\":\"2010\",\"pages\":\"167-169\",\"volume\":\"4\",\"id\":\"032a0fa0-83ef-3610-bdcc-94e56a23bb75\",\"created\":\"2023-01-10T01:45:23.480Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:23.480Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.\",\"bibtype\":\"article\",\"author\":\"Hynson, R.M.G. and Kwan, A.H. and Jacques, D.A. and MacKay, J.P. and Trewhella, J.\",\"doi\":\"10.1007/s12104-010-9237-6\",\"journal\":\"Biomolecular NMR Assignments\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis},\\n type = {article},\\n year = {2010},\\n pages = {167-169},\\n volume = {4},\\n id = {032a0fa0-83ef-3610-bdcc-94e56a23bb75},\\n created = {2023-01-10T01:45:23.480Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:23.480Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.},\\n bibtype = {article},\\n author = {Hynson, R.M.G. and Kwan, A.H. and Jacques, D.A. and MacKay, J.P. and Trewhella, J.},\\n doi = {10.1007/s12104-010-9237-6},\\n journal = {Biomolecular NMR Assignments},\\n number = {2}\\n}\",\"author_short\":[\"Hynson,  R.\",\"Kwan,  A.\",\"Jacques,  D.\",\"MacKay,  J.\",\"Trewhella,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"hynson-kwan-jacques-mackay-trewhella-sup1suphsup13supcandsup15supnbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Protein Recognition\",\"type\":\"book\",\"year\":\"2010\",\"source\":\"Amino Acids, Peptides and Proteins in Organic Chemistry\",\"pages\":\"505-532\",\"volume\":\"2\",\"id\":\"cf609a26-bd7d-39ca-a12c-7a19ab224d44\",\"created\":\"2023-01-10T01:45:24.389Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:24.389Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"book\",\"author\":\"Mansfield, R.E. and Cross, A.J. and Matthews, J.M. and Mackay, J.P.\",\"doi\":\"10.1002/9783527631780.ch12\",\"bibtex\":\"@book{\\n title = {Protein Recognition},\\n type = {book},\\n year = {2010},\\n source = {Amino Acids, Peptides and Proteins in Organic Chemistry},\\n pages = {505-532},\\n volume = {2},\\n id = {cf609a26-bd7d-39ca-a12c-7a19ab224d44},\\n created = {2023-01-10T01:45:24.389Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:24.389Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {book},\\n author = {Mansfield, R.E. and Cross, A.J. and Matthews, J.M. and Mackay, J.P.},\\n doi = {10.1002/9783527631780.ch12}\\n}\",\"author_short\":[\"Mansfield,  R.\",\"Cross,  A.\",\"Matthews,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mansfield-cross-matthews-mackay-proteinrecognition-2010\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The structural analysis of protein-protein interactions by NMR spectroscopy\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"5224-5232\",\"volume\":\"9\",\"id\":\"388e096a-15bd-345c-96eb-0a1702c4a9b6\",\"created\":\"2023-01-10T01:45:25.437Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:25.437Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.\",\"bibtype\":\"article\",\"author\":\"O'Connell, M.R. and Gamsjaeger, R. and Mackay, J.P.\",\"doi\":\"10.1002/pmic.200900303\",\"journal\":\"Proteomics\",\"number\":\"23\",\"bibtex\":\"@article{\\n title = {The structural analysis of protein-protein interactions by NMR spectroscopy},\\n type = {article},\\n year = {2009},\\n pages = {5224-5232},\\n volume = {9},\\n id = {388e096a-15bd-345c-96eb-0a1702c4a9b6},\\n created = {2023-01-10T01:45:25.437Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:25.437Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.},\\n bibtype = {article},\\n author = {O'Connell, M.R. and Gamsjaeger, R. and Mackay, J.P.},\\n doi = {10.1002/pmic.200900303},\\n journal = {Proteomics},\\n number = {23}\\n}\",\"author_short\":[\"O'Connell,  M.\",\"Gamsjaeger,  R.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"oconnell-gamsjaeger-mackay-thestructuralanalysisofproteinproteininteractionsbynmrspectroscopy-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Conformational Stability and DNA Binding Specificity of the Cardiac T-Box Transcription Factor Tbx20\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"606-618\",\"volume\":\"389\",\"id\":\"c62d0b72-ffae-3a09-8c2e-63a5222cc5a2\",\"created\":\"2023-01-10T01:45:26.344Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:26.344Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The transcription factor Tbx20 acts within a hierarchy of T-box factors in lineage specification and morphogenesis in the mammalian heart and is mutated in congenital heart disease. T-box family members share a ∼ 20-kDa DNA-binding domain termed the T-box. The question of how highly homologous T-box proteins achieve differential transcriptional control in heart development, while apparently binding to the same DNA sequence, remains unresolved. Here we show that the optimal DNA recognition sequence for the T-box of Tbx20 corresponds to a T-half-site. Furthermore, we demonstrate using purified recombinant domains that distinct T-boxes show significant differences in the affinity and kinetics of binding and in conformational stability, with the T-box of Tbx20 displaying molten globule character. Our data highlight unique features of Tbx20 and suggest mechanistic ways in which cardiac T-box factors might interact synergistically and/or competitively within the cardiac regulatory network. © 2009 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Macindoe, I. and Glockner, L. and Vukašin, P. and Stennard, F.A. and Costa, M.W. and Harvey, R.P. and Mackay, J.P. and Sunde, M.\",\"doi\":\"10.1016/j.jmb.2009.04.056\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Conformational Stability and DNA Binding Specificity of the Cardiac T-Box Transcription Factor Tbx20},\\n type = {article},\\n year = {2009},\\n pages = {606-618},\\n volume = {389},\\n id = {c62d0b72-ffae-3a09-8c2e-63a5222cc5a2},\\n created = {2023-01-10T01:45:26.344Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:26.344Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The transcription factor Tbx20 acts within a hierarchy of T-box factors in lineage specification and morphogenesis in the mammalian heart and is mutated in congenital heart disease. T-box family members share a ∼ 20-kDa DNA-binding domain termed the T-box. The question of how highly homologous T-box proteins achieve differential transcriptional control in heart development, while apparently binding to the same DNA sequence, remains unresolved. Here we show that the optimal DNA recognition sequence for the T-box of Tbx20 corresponds to a T-half-site. Furthermore, we demonstrate using purified recombinant domains that distinct T-boxes show significant differences in the affinity and kinetics of binding and in conformational stability, with the T-box of Tbx20 displaying molten globule character. Our data highlight unique features of Tbx20 and suggest mechanistic ways in which cardiac T-box factors might interact synergistically and/or competitively within the cardiac regulatory network. © 2009 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Macindoe, I. and Glockner, L. and Vukašin, P. and Stennard, F.A. and Costa, M.W. and Harvey, R.P. and Mackay, J.P. and Sunde, M.},\\n doi = {10.1016/j.jmb.2009.04.056},\\n journal = {Journal of Molecular Biology},\\n number = {3}\\n}\",\"author_short\":[\"Macindoe,  I.\",\"Glockner,  L.\",\"Vukašin,  P.\",\"Stennard,  F.\",\"Costa,  M.\",\"Harvey,  R.\",\"Mackay,  J.\",\"Sunde,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"macindoe-glockner-vukain-stennard-costa-harvey-mackay-sunde-conformationalstabilityanddnabindingspecificityofthecardiactboxtranscriptionfactortbx20-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"29462-29469\",\"volume\":\"284\",\"id\":\"a280cdd1-9f7b-37f1-8e93-e668c33b7b99\",\"created\":\"2023-01-10T01:45:27.256Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:27.256Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"α-Hemoglobin (αHb) stabilizing protein (AHSP) is expressed in erythropoietic tissues as an accessory factor in hemoglobin synthesis. AHSP forms a specific complex with αHb and suppresses the heme-catalyzed evolution of reactive oxygen species by converting αHb to a conformation in which the heme is coordinated at both axial positions by histidine side chains (bis-histidyl coordination). Currently, the detailed mechanism by which AHSP induces structural changes in αHb has not been determined. Here, we present x-ray crystallography, NMR spectroscopy, and mutagenesis data that identify, for the first time, the importance of an evolutionarily conserved proline, Pro30, in loop 1 of AHSP. Mutation of Pro30 to a variety of residue types results in reduced ability to convert αHb. In complex with-Hb, AHSP Pro30 adopts a cis-peptidyl conformation and makes contact with the N terminus of helix G in αHb. Mutations that stabilize the cis-peptidyl conformation of free AHSP, also enhance the αHb conversion activity. These findings suggest that AHSP loop 1 can transmit structural changes to the heme pocket of αHb, and, more generally, highlight the importance of cis-peptidyl prolyl residues in defining the conformation of regulatory protein loops. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Gell, D.A. and Feng, L. and Zhou, S. and Jeffrey, P.D. and Bendak, K. and Gow, A. and Weiss, M.J. and Shi, Y. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M109.027045\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"43\",\"bibtex\":\"@article{\\n title = {A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin},\\n type = {article},\\n year = {2009},\\n pages = {29462-29469},\\n volume = {284},\\n id = {a280cdd1-9f7b-37f1-8e93-e668c33b7b99},\\n created = {2023-01-10T01:45:27.256Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:27.256Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {α-Hemoglobin (αHb) stabilizing protein (AHSP) is expressed in erythropoietic tissues as an accessory factor in hemoglobin synthesis. AHSP forms a specific complex with αHb and suppresses the heme-catalyzed evolution of reactive oxygen species by converting αHb to a conformation in which the heme is coordinated at both axial positions by histidine side chains (bis-histidyl coordination). Currently, the detailed mechanism by which AHSP induces structural changes in αHb has not been determined. Here, we present x-ray crystallography, NMR spectroscopy, and mutagenesis data that identify, for the first time, the importance of an evolutionarily conserved proline, Pro30, in loop 1 of AHSP. Mutation of Pro30 to a variety of residue types results in reduced ability to convert αHb. In complex with-Hb, AHSP Pro30 adopts a cis-peptidyl conformation and makes contact with the N terminus of helix G in αHb. Mutations that stabilize the cis-peptidyl conformation of free AHSP, also enhance the αHb conversion activity. These findings suggest that AHSP loop 1 can transmit structural changes to the heme pocket of αHb, and, more generally, highlight the importance of cis-peptidyl prolyl residues in defining the conformation of regulatory protein loops. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Gell, D.A. and Feng, L. and Zhou, S. and Jeffrey, P.D. and Bendak, K. and Gow, A. and Weiss, M.J. and Shi, Y. and Mackay, J.P.},\\n doi = {10.1074/jbc.M109.027045},\\n journal = {Journal of Biological Chemistry},\\n number = {43}\\n}\",\"author_short\":[\"Gell,  D.\",\"Feng,  L.\",\"Zhou,  S.\",\"Jeffrey,  P.\",\"Bendak,  K.\",\"Gow,  A.\",\"Weiss,  M.\",\"Shi,  Y.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gell-feng-zhou-jeffrey-bendak-gow-weiss-shi-etal-acisprolineinhemoglobinstabilizingproteindirectsthestructuralreorganizationofhemoglobin-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"179-187\",\"volume\":\"423\",\"id\":\"852e680d-f3c3-3677-a4d0-54c4e5f30958\",\"created\":\"2023-01-10T01:45:28.180Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:28.180Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation ormethylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2-H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin. © The Authors Journal compilation. © 2009 Biochemical Society.\",\"bibtype\":\"article\",\"author\":\"Musselman, C.A. and Mansfield, R.E. and Garske, A.L. and Davrazou, F. and Kwan, A.H. and Oliver, S.S. and O'Leary, H. and Denu, J.M. and Mackay, J.P. and Kutateladze, T.G.\",\"doi\":\"10.1042/BJ20090870\",\"journal\":\"Biochemical Journal\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications},\\n type = {article},\\n year = {2009},\\n pages = {179-187},\\n volume = {423},\\n id = {852e680d-f3c3-3677-a4d0-54c4e5f30958},\\n created = {2023-01-10T01:45:28.180Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:28.180Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation ormethylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2-H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin. © The Authors Journal compilation. © 2009 Biochemical Society.},\\n bibtype = {article},\\n author = {Musselman, C.A. and Mansfield, R.E. and Garske, A.L. and Davrazou, F. and Kwan, A.H. and Oliver, S.S. and O'Leary, H. and Denu, J.M. and Mackay, J.P. and Kutateladze, T.G.},\\n doi = {10.1042/BJ20090870},\\n journal = {Biochemical Journal},\\n number = {2}\\n}\",\"author_short\":[\"Musselman,  C.\",\"Mansfield,  R.\",\"Garske,  A.\",\"Davrazou,  F.\",\"Kwan,  A.\",\"Oliver,  S.\",\"O'Leary,  H.\",\"Denu,  J.\",\"Mackay,  J.\",\"Kutateladze,  T.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Erratum: Structure and inhibition of orotidine 5#-Monophosphate decarboxylase from plasmodium falciparum (Biochemistry (2009) 48:11 (2570-2570) 10.1021/bi900043p)\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"2570\",\"volume\":\"48\",\"id\":\"600819d9-be33-3029-9db3-d7575a665191\",\"created\":\"2023-01-10T01:45:29.095Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:29.095Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Langley, D.B. and Shojaei, M. and Chan, C. and Lok, H.C. and Mackay, J.P. and Traut, T.W. and Guss, J.M. and Christopherson, R.I.\",\"doi\":\"10.1021/bi900043p\",\"journal\":\"Biochemistry\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Erratum: Structure and inhibition of orotidine 5#-Monophosphate decarboxylase from plasmodium falciparum (Biochemistry (2009) 48:11 (2570-2570) 10.1021/bi900043p)},\\n type = {article},\\n year = {2009},\\n pages = {2570},\\n volume = {48},\\n id = {600819d9-be33-3029-9db3-d7575a665191},\\n created = {2023-01-10T01:45:29.095Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:29.095Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Langley, D.B. and Shojaei, M. and Chan, C. and Lok, H.C. and Mackay, J.P. and Traut, T.W. and Guss, J.M. and Christopherson, R.I.},\\n doi = {10.1021/bi900043p},\\n journal = {Biochemistry},\\n number = {11}\\n}\",\"author_short\":[\"Langley,  D.\",\"Shojaei,  M.\",\"Chan,  C.\",\"Lok,  H.\",\"Mackay,  J.\",\"Traut,  T.\",\"Guss,  J.\",\"Christopherson,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"langley-shojaei-chan-lok-mackay-traut-guss-christopherson-erratumstructureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparumbiochemistry2009481125702570101021bi900043p-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"A mass spectrometric investigation of the ability of metal complexes to modulate transcription factor activity\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"5546-5548\",\"id\":\"674f52e4-6615-311d-9893-ba1bd7e3946c\",\"created\":\"2023-01-10T01:45:30.041Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:30.041Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"ESI mass spectrometry was used to assess the ability of metal complexes to inhibit binding of a transcription factor to a DNA molecule containing its recognition sequence. © 2009 The Royal Society of Chemistry.\",\"bibtype\":\"article\",\"author\":\"Talib, J. and Beck, J.L. and Urathamakul, T. and Nguyen, C.D. and Aldrich-Wright, J.R. and MacKay, J.P. and Ralph, S.F.\",\"doi\":\"10.1039/b904751d\",\"journal\":\"Chemical Communications\",\"number\":\"37\",\"bibtex\":\"@article{\\n title = {A mass spectrometric investigation of the ability of metal complexes to modulate transcription factor activity},\\n type = {article},\\n year = {2009},\\n pages = {5546-5548},\\n id = {674f52e4-6615-311d-9893-ba1bd7e3946c},\\n created = {2023-01-10T01:45:30.041Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:30.041Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {ESI mass spectrometry was used to assess the ability of metal complexes to inhibit binding of a transcription factor to a DNA molecule containing its recognition sequence. © 2009 The Royal Society of Chemistry.},\\n bibtype = {article},\\n author = {Talib, J. and Beck, J.L. and Urathamakul, T. and Nguyen, C.D. and Aldrich-Wright, J.R. and MacKay, J.P. and Ralph, S.F.},\\n doi = {10.1039/b904751d},\\n journal = {Chemical Communications},\\n number = {37}\\n}\",\"author_short\":[\"Talib,  J.\",\"Beck,  J.\",\"Urathamakul,  T.\",\"Nguyen,  C.\",\"Aldrich-Wright,  J.\",\"MacKay,  J.\",\"Ralph,  S.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"talib-beck-urathamakul-nguyen-aldrichwright-mackay-ralph-amassspectrometricinvestigationoftheabilityofmetalcomplexestomodulatetranscriptionfactoractivity-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural analysis of MED-1 reveals unexpected diversity in the mechanism of DNA recognition by GATA-type zinc finger domains\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"5827-5835\",\"volume\":\"284\",\"id\":\"71df5cc8-308b-387c-9226-824ae9beba6f\",\"created\":\"2023-01-10T01:45:30.973Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:30.973Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"MED-1 is a member of a group of divergent GATA-type zinc finger proteins recently identified in several species of Caenorhabditis. The med genes are transcriptional regulators that are involved in the specification of the mesoderm and endoderm precursor cells in nematodes. Unlike other GATA-type zinc fingers that recognize the consensus sequence (A/C/ T)GATA(A/G), the MED-1 zinc finger (MED1zf) binds the larger and atypical site GTATACT(T/C)3. We have examined the basis for this unusual DNA specificity using a range of biochemical and biophysical approaches. Most strikingly, we show that although the core of the MED1zf structure is similar to that of GATA-1, the basic tail C-terminal to the zinc finger unexpectedly adopts an α-helical structure upon binding DNA. This additional helix appears to contact the major groove of the DNA, making contacts that explain the extended DNA consensus sequence observed for MED1zf. Our data expand the versatility of DNA recognition by GATA-type zinc fingers and perhaps shed new light on the DNA-binding properties of mammalian GATA factors. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Lowry, J.A. and Gamsjaeger, R. and Thong, S.Y. and Hung, W. and Kwan, A.H. and Broitman-Maduro, G. and Matthews, J.M. and Maduro, M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M808712200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {Structural analysis of MED-1 reveals unexpected diversity in the mechanism of DNA recognition by GATA-type zinc finger domains},\\n type = {article},\\n year = {2009},\\n pages = {5827-5835},\\n volume = {284},\\n id = {71df5cc8-308b-387c-9226-824ae9beba6f},\\n created = {2023-01-10T01:45:30.973Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:30.973Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {MED-1 is a member of a group of divergent GATA-type zinc finger proteins recently identified in several species of Caenorhabditis. The med genes are transcriptional regulators that are involved in the specification of the mesoderm and endoderm precursor cells in nematodes. Unlike other GATA-type zinc fingers that recognize the consensus sequence (A/C/ T)GATA(A/G), the MED-1 zinc finger (MED1zf) binds the larger and atypical site GTATACT(T/C)3. We have examined the basis for this unusual DNA specificity using a range of biochemical and biophysical approaches. Most strikingly, we show that although the core of the MED1zf structure is similar to that of GATA-1, the basic tail C-terminal to the zinc finger unexpectedly adopts an α-helical structure upon binding DNA. This additional helix appears to contact the major groove of the DNA, making contacts that explain the extended DNA consensus sequence observed for MED1zf. Our data expand the versatility of DNA recognition by GATA-type zinc fingers and perhaps shed new light on the DNA-binding properties of mammalian GATA factors. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Lowry, J.A. and Gamsjaeger, R. and Thong, S.Y. and Hung, W. and Kwan, A.H. and Broitman-Maduro, G. and Matthews, J.M. and Maduro, M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M808712200},\\n journal = {Journal of Biological Chemistry},\\n number = {9}\\n}\",\"author_short\":[\"Lowry,  J.\",\"Gamsjaeger,  R.\",\"Thong,  S.\",\"Hung,  W.\",\"Kwan,  A.\",\"Broitman-Maduro,  G.\",\"Matthews,  J.\",\"Maduro,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"lowry-gamsjaeger-thong-hung-kwan-broitmanmaduro-matthews-maduro-etal-structuralanalysisofmed1revealsunexpecteddiversityinthemechanismofdnarecognitionbygatatypezincfingerdomains-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The zinc fingers of the SR-like protein ZRANB2 are single-stranded RNA-binding domains that recognize 5' splice site-like sequences\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"5581-5586\",\"volume\":\"106\",\"id\":\"bbb64689-b326-3416-95e2-8b6b88f2d5e6\",\"created\":\"2023-01-10T01:45:31.914Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:31.914Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The alternative splicing of mRNA is a critical process in higher eukaryotes that generates substantial proteomic diversity. Many of the proteins that are essential to this process contain arginine/serine-rich (RS) domains. ZRANB2 is a widely-expressed and highly-conserved RS-domain protein that can regulate alternative splicing but lacks canonical RNA-binding domains. Instead, it contains 2 RanBP2-type zinc finger (ZnF) domains. We demonstrate that these ZnFs recognize ssRNA with high affinity and specificity. Each ZnF binds to a single AGGUAA motif and the 2 domains combine to recognize AGGUAA (Nx) AGGUAA double sites, suggesting that ZRANB2 regulates alternative splicing via a direct interaction with pre-mRNA at sites that resemble the consensus 5' splice site. We show using X-ray crystallography that recognition of an AGGUAA motif by a single ZnF is dominated by side-chain hydrogen bonds to the bases and formation of a guanine-tryptophan-guanine ''ladder.'' A number of other human proteins that function in RNA processing also contain RanBP2 ZnFs in which the RNA-binding residues of ZRANB2 are conserved. The ZnFs of ZRANB2 therefore define another class of RNA-binding domain, advancing our understanding of RNA recognition and emphasizing the versatility of domains in molecular recognition.\",\"bibtype\":\"article\",\"author\":\"Loughlin, F.E. and Mansfield, R.E. and Vaz, P.M. and McGrath, A.P. and Setiyaputra, S. and Gamsjaeger, R. and Chen, E.S. and Morris, B.J. and Guss, J.M. and Mackay, J.P.\",\"doi\":\"10.1073/pnas.0802466106\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"14\",\"bibtex\":\"@article{\\n title = {The zinc fingers of the SR-like protein ZRANB2 are single-stranded RNA-binding domains that recognize 5' splice site-like sequences},\\n type = {article},\\n year = {2009},\\n pages = {5581-5586},\\n volume = {106},\\n id = {bbb64689-b326-3416-95e2-8b6b88f2d5e6},\\n created = {2023-01-10T01:45:31.914Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:31.914Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The alternative splicing of mRNA is a critical process in higher eukaryotes that generates substantial proteomic diversity. Many of the proteins that are essential to this process contain arginine/serine-rich (RS) domains. ZRANB2 is a widely-expressed and highly-conserved RS-domain protein that can regulate alternative splicing but lacks canonical RNA-binding domains. Instead, it contains 2 RanBP2-type zinc finger (ZnF) domains. We demonstrate that these ZnFs recognize ssRNA with high affinity and specificity. Each ZnF binds to a single AGGUAA motif and the 2 domains combine to recognize AGGUAA (Nx) AGGUAA double sites, suggesting that ZRANB2 regulates alternative splicing via a direct interaction with pre-mRNA at sites that resemble the consensus 5' splice site. We show using X-ray crystallography that recognition of an AGGUAA motif by a single ZnF is dominated by side-chain hydrogen bonds to the bases and formation of a guanine-tryptophan-guanine ''ladder.'' A number of other human proteins that function in RNA processing also contain RanBP2 ZnFs in which the RNA-binding residues of ZRANB2 are conserved. The ZnFs of ZRANB2 therefore define another class of RNA-binding domain, advancing our understanding of RNA recognition and emphasizing the versatility of domains in molecular recognition.},\\n bibtype = {article},\\n author = {Loughlin, F.E. and Mansfield, R.E. and Vaz, P.M. and McGrath, A.P. and Setiyaputra, S. and Gamsjaeger, R. and Chen, E.S. and Morris, B.J. and Guss, J.M. and Mackay, J.P.},\\n doi = {10.1073/pnas.0802466106},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {14}\\n}\",\"author_short\":[\"Loughlin,  F.\",\"Mansfield,  R.\",\"Vaz,  P.\",\"McGrath,  A.\",\"Setiyaputra,  S.\",\"Gamsjaeger,  R.\",\"Chen,  E.\",\"Morris,  B.\",\"Guss,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"loughlin-mansfield-vaz-mcgrath-setiyaputra-gamsjaeger-chen-morris-etal-thezincfingersofthesrlikeproteinzranb2aresinglestrandedrnabindingdomainsthatrecognize5splicesitelikesequences-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"It takes two to tango: The structure and function of LIM, RING, PHD and MYND domains\",\"type\":\"article\",\"year\":\"2009\",\"pages\":\"3681-3696\",\"volume\":\"15\",\"id\":\"12e1371d-f8f7-3ef4-8a33-189b9e2b23cc\",\"created\":\"2023-01-10T01:45:32.880Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:32.880Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"LIM (Lin-11, Isl-1, Mec-3), RING (Really interesting new gene), PHD (Plant homology domain) and MYND (myeloid, Nervy, DEAF-1) domains are all zinc-binding domains that ligate two zinc ions. Unlike the better known classical zinc fingers, these domains do not bind DNA, but instead mediate interactions with other proteins. LIM-domain containing proteins have diverse functions as regulators of gene expression, cell adhesion and motility and signal transduction. RING finger proteins are generally associated with ubiquitination; the presence of such a domain is the defining feature of a class of E3 ubiquitin protein ligases. PHD proteins have been associated with SUMOylation but most recently have emerged as a chromatin recognition motif that reads the methylation state of histones. The function of the MYND domain is less clear, but MYND domains are also found in proteins that have ubiquitin ligase and/or histone methyltransferase activity. Here we review the structure-function relationships for these domains and discuss strategies to modulate their activity. © 2009 Bentham Science Publishers Ltd.\",\"bibtype\":\"article\",\"author\":\"Matthews, J.M. and Bhati, M. and Lehtomaki, E. and Mansfield, R.E. and Cubeddu, L. and Mackay, J.P.\",\"doi\":\"10.2174/138161209789271861\",\"journal\":\"Current Pharmaceutical Design\",\"number\":\"31\",\"bibtex\":\"@article{\\n title = {It takes two to tango: The structure and function of LIM, RING, PHD and MYND domains},\\n type = {article},\\n year = {2009},\\n pages = {3681-3696},\\n volume = {15},\\n id = {12e1371d-f8f7-3ef4-8a33-189b9e2b23cc},\\n created = {2023-01-10T01:45:32.880Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:32.880Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {LIM (Lin-11, Isl-1, Mec-3), RING (Really interesting new gene), PHD (Plant homology domain) and MYND (myeloid, Nervy, DEAF-1) domains are all zinc-binding domains that ligate two zinc ions. Unlike the better known classical zinc fingers, these domains do not bind DNA, but instead mediate interactions with other proteins. LIM-domain containing proteins have diverse functions as regulators of gene expression, cell adhesion and motility and signal transduction. RING finger proteins are generally associated with ubiquitination; the presence of such a domain is the defining feature of a class of E3 ubiquitin protein ligases. PHD proteins have been associated with SUMOylation but most recently have emerged as a chromatin recognition motif that reads the methylation state of histones. The function of the MYND domain is less clear, but MYND domains are also found in proteins that have ubiquitin ligase and/or histone methyltransferase activity. Here we review the structure-function relationships for these domains and discuss strategies to modulate their activity. © 2009 Bentham Science Publishers Ltd.},\\n bibtype = {article},\\n author = {Matthews, J.M. and Bhati, M. and Lehtomaki, E. and Mansfield, R.E. and Cubeddu, L. and Mackay, J.P.},\\n doi = {10.2174/138161209789271861},\\n journal = {Current Pharmaceutical Design},\\n number = {31}\\n}\",\"author_short\":[\"Matthews,  J.\",\"Bhati,  M.\",\"Lehtomaki,  E.\",\"Mansfield,  R.\",\"Cubeddu,  L.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"matthews-bhati-lehtomaki-mansfield-cubeddu-mackay-ittakestwototangothestructureandfunctionoflimringphdandmynddomains-2009\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"NMR spectroscopy as a tool for the rapid assessment of the conformation of GST-fusion proteins\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"1630-1635\",\"volume\":\"17\",\"id\":\"2555f698-f625-3490-b864-b58850f3abab\",\"created\":\"2023-01-10T01:45:33.906Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:33.906Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Glutathione-S-transferase (GST)-fusion proteins are used extensively for structural, biochemical, and functional analyses. Although the conformation of the target protein is of critical importance, confirmation of the folded state of the target is often not undertaken or is cumbersome because of the requirement to first remove the GST tag. Here, we demonstrate that it is possible to record conventional 15N-HSQC NMR spectra of small GST-fusion proteins and that the observed signals arise almost exclusively from the target protein. This approach constitutes a rapid and straightforward means of assessing the conformation of a GST-fusion protein without having to cleave the GST and should prove valuable, both to biochemists seeking to check the conformation of their proteins prior to functional studies and to structural biologists screening protein constructs for suitability as targets for structural studies. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 The Protein Society.\",\"bibtype\":\"article\",\"author\":\"Chu, K.L. and Gamsjaeger, R. and Mansfield, R.E. and Mackay, J.P.\",\"doi\":\"10.1110/ps.034983.108\",\"journal\":\"Protein Science\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {NMR spectroscopy as a tool for the rapid assessment of the conformation of GST-fusion proteins},\\n type = {article},\\n year = {2008},\\n pages = {1630-1635},\\n volume = {17},\\n id = {2555f698-f625-3490-b864-b58850f3abab},\\n created = {2023-01-10T01:45:33.906Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:33.906Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Glutathione-S-transferase (GST)-fusion proteins are used extensively for structural, biochemical, and functional analyses. Although the conformation of the target protein is of critical importance, confirmation of the folded state of the target is often not undertaken or is cumbersome because of the requirement to first remove the GST tag. Here, we demonstrate that it is possible to record conventional 15N-HSQC NMR spectra of small GST-fusion proteins and that the observed signals arise almost exclusively from the target protein. This approach constitutes a rapid and straightforward means of assessing the conformation of a GST-fusion protein without having to cleave the GST and should prove valuable, both to biochemists seeking to check the conformation of their proteins prior to functional studies and to structural biologists screening protein constructs for suitability as targets for structural studies. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 The Protein Society.},\\n bibtype = {article},\\n author = {Chu, K.L. and Gamsjaeger, R. and Mansfield, R.E. and Mackay, J.P.},\\n doi = {10.1110/ps.034983.108},\\n journal = {Protein Science},\\n number = {9}\\n}\",\"author_short\":[\"Chu,  K.\",\"Gamsjaeger,  R.\",\"Mansfield,  R.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"chu-gamsjaeger-mansfield-mackay-nmrspectroscopyasatoolfortherapidassessmentoftheconformationofgstfusionproteins-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural analysis of hydrophobins\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"773-784\",\"volume\":\"39\",\"id\":\"d6614c6a-3917-3fb1-bfca-b4688d016cf4\",\"created\":\"2023-01-10T01:45:34.809Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:34.809Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Hydrophobins are a remarkable class of small cysteine-rich proteins found exclusively in fungi. They self-assemble to form robust polymeric monolayers that are highly amphipathic and play numerous roles in fungal biology, such as in the formation and dispersal of aerial spores and in pathogenic and mutualistic interactions. The polymeric form can be reversibly disassembled and is able to reverse the wettability of a surface, leading to many proposals for nanotechnological applications over recent years. The surprising properties of hydrophobins and their potential for commercialization have led to substantial efforts to delineate their morphology and molecular structure. In this review, we summarize the progress that has been made using a variety of spectroscopic and microscopic approaches towards understanding the molecular mechanisms underlying hydrophobin structure. © 2007 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Sunde, M. and Kwan, A.H.Y. and Templeton, M.D. and Beever, R.E. and Mackay, J.P.\",\"doi\":\"10.1016/j.micron.2007.08.003\",\"journal\":\"Micron\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {Structural analysis of hydrophobins},\\n type = {article},\\n year = {2008},\\n pages = {773-784},\\n volume = {39},\\n id = {d6614c6a-3917-3fb1-bfca-b4688d016cf4},\\n created = {2023-01-10T01:45:34.809Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:34.809Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Hydrophobins are a remarkable class of small cysteine-rich proteins found exclusively in fungi. They self-assemble to form robust polymeric monolayers that are highly amphipathic and play numerous roles in fungal biology, such as in the formation and dispersal of aerial spores and in pathogenic and mutualistic interactions. The polymeric form can be reversibly disassembled and is able to reverse the wettability of a surface, leading to many proposals for nanotechnological applications over recent years. The surprising properties of hydrophobins and their potential for commercialization have led to substantial efforts to delineate their morphology and molecular structure. In this review, we summarize the progress that has been made using a variety of spectroscopic and microscopic approaches towards understanding the molecular mechanisms underlying hydrophobin structure. © 2007 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Sunde, M. and Kwan, A.H.Y. and Templeton, M.D. and Beever, R.E. and Mackay, J.P.},\\n doi = {10.1016/j.micron.2007.08.003},\\n journal = {Micron},\\n number = {7}\\n}\",\"author_short\":[\"Sunde,  M.\",\"Kwan,  A.\",\"Templeton,  M.\",\"Beever,  R.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"sunde-kwan-templeton-beever-mackay-structuralanalysisofhydrophobins-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The Cys3-Cys4 Loop of the Hydrophobin EAS Is Not Required for Rodlet Formation and Surface Activity\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"708-720\",\"volume\":\"382\",\"id\":\"209bc497-4431-3395-a923-3ba1135b8fac\",\"created\":\"2023-01-10T01:45:35.837Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:35.837Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Class I hydrophobins are fungal proteins that self-assemble into robust amphipathic rodlet monolayers on the surface of aerial structures such as spores and fruiting bodies. These layers share many structural characteristics with amyloid fibrils and belong to the growing family of functional amyloid-like materials produced by microorganisms. Although the three-dimensional structure of the soluble monomeric form of a class I hydrophobin has been determined, little is known about the molecular structure of the rodlets or their assembly mechanism. Several models have been proposed, some of which suggest that the Cys3-Cys4 loop has a critical role in the initiation of assembly or in the polymeric structure. In order to provide insight into the relationship between hydrophobin sequence and rodlet assembly, we investigated the role of the Cys3-Cys4 loop in EAS, a class I hydrophobin from Neurospora crassa. Remarkably, deletion of up to 15 residues from this 25-residue loop does not impair rodlet formation or reduce the surface activity of the protein, and the physicochemical properties of rodlets formed by this mutant are indistinguishable from those of its full-length counterpart. In addition, the core structure of the truncation mutant is essentially unchanged. Molecular dynamics simulations carried out on the full-length protein and this truncation mutant binding to an air-water interface show that, although it is hydrophobic, the loop does not play a role in positioning the protein at the surface. These results demonstrate that the Cys3-Cys4 loop does not have an integral role in the formation or structure of the rodlets and that the major determinant of the unique properties of these proteins is the amphipathic core structure, which is likely to be preserved in all hydrophobins despite the high degree of sequence variation across the family. © 2008 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Kwan, A.H. and Macindoe, I. and Vukašin, P.V. and Morris, V.K. and Kass, I. and Gupte, R. and Mark, A.E. and Templeton, M.D. and Mackay, J.P. and Sunde, M.\",\"doi\":\"10.1016/j.jmb.2008.07.034\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {The Cys3-Cys4 Loop of the Hydrophobin EAS Is Not Required for Rodlet Formation and Surface Activity},\\n type = {article},\\n year = {2008},\\n pages = {708-720},\\n volume = {382},\\n id = {209bc497-4431-3395-a923-3ba1135b8fac},\\n created = {2023-01-10T01:45:35.837Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:35.837Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Class I hydrophobins are fungal proteins that self-assemble into robust amphipathic rodlet monolayers on the surface of aerial structures such as spores and fruiting bodies. These layers share many structural characteristics with amyloid fibrils and belong to the growing family of functional amyloid-like materials produced by microorganisms. Although the three-dimensional structure of the soluble monomeric form of a class I hydrophobin has been determined, little is known about the molecular structure of the rodlets or their assembly mechanism. Several models have been proposed, some of which suggest that the Cys3-Cys4 loop has a critical role in the initiation of assembly or in the polymeric structure. In order to provide insight into the relationship between hydrophobin sequence and rodlet assembly, we investigated the role of the Cys3-Cys4 loop in EAS, a class I hydrophobin from Neurospora crassa. Remarkably, deletion of up to 15 residues from this 25-residue loop does not impair rodlet formation or reduce the surface activity of the protein, and the physicochemical properties of rodlets formed by this mutant are indistinguishable from those of its full-length counterpart. In addition, the core structure of the truncation mutant is essentially unchanged. Molecular dynamics simulations carried out on the full-length protein and this truncation mutant binding to an air-water interface show that, although it is hydrophobic, the loop does not play a role in positioning the protein at the surface. These results demonstrate that the Cys3-Cys4 loop does not have an integral role in the formation or structure of the rodlets and that the major determinant of the unique properties of these proteins is the amphipathic core structure, which is likely to be preserved in all hydrophobins despite the high degree of sequence variation across the family. © 2008 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Kwan, A.H. and Macindoe, I. and Vukašin, P.V. and Morris, V.K. and Kass, I. and Gupte, R. and Mark, A.E. and Templeton, M.D. and Mackay, J.P. and Sunde, M.},\\n doi = {10.1016/j.jmb.2008.07.034},\\n journal = {Journal of Molecular Biology},\\n number = {3}\\n}\",\"author_short\":[\"Kwan,  A.\",\"Macindoe,  I.\",\"Vukašin,  P.\",\"Morris,  V.\",\"Kass,  I.\",\"Gupte,  R.\",\"Mark,  A.\",\"Templeton,  M.\",\"Mackay,  J.\",\"Sunde,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kwan-macindoe-vukain-morris-kass-gupte-mark-templeton-etal-thecys3cys4loopofthehydrophobineasisnotrequiredforrodletformationandsurfaceactivity-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Response to Chatr-aryamontri et al.: Protein interactions: to believe or not to believe?\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"242-243\",\"volume\":\"33\",\"id\":\"a93b20e9-5f78-368e-9bc7-126a1f8e9c4e\",\"created\":\"2023-01-10T01:45:36.806Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:36.806Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Sunde, M. and Lowry, J.A. and Crossley, M. and Matthews, J.M.\",\"doi\":\"10.1016/j.tibs.2008.04.003\",\"journal\":\"Trends in Biochemical Sciences\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Response to Chatr-aryamontri et al.: Protein interactions: to believe or not to believe?},\\n type = {article},\\n year = {2008},\\n pages = {242-243},\\n volume = {33},\\n id = {a93b20e9-5f78-368e-9bc7-126a1f8e9c4e},\\n created = {2023-01-10T01:45:36.806Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:36.806Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Mackay, J.P. and Sunde, M. and Lowry, J.A. and Crossley, M. and Matthews, J.M.},\\n doi = {10.1016/j.tibs.2008.04.003},\\n journal = {Trends in Biochemical Sciences},\\n number = {6}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Sunde,  M.\",\"Lowry,  J.\",\"Crossley,  M.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-sunde-lowry-crossley-matthews-responsetochatraryamontrietalproteininteractionstobelieveornottobelieve-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Evolution of Quaternary Structure in a Homotetrameric Enzyme\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"691-703\",\"volume\":\"380\",\"id\":\"63b6023f-2683-3fd8-a96d-6ee13c0b2a51\",\"created\":\"2023-01-10T01:45:37.717Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:37.717Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Dihydrodipicolinate synthase (DHDPS) is an essential enzyme in (S)-lysine biosynthesis and an important antibiotic target. All X-ray crystal structures solved to date reveal a homotetrameric enzyme. In order to explore the role of this quaternary structure, dimeric variants of Escherichia coli DHDPS were engineered and their properties were compared to those of the wild-type tetrameric form. X-ray crystallography reveals that the active site is not disturbed when the quaternary structure is disrupted. However, the activity of the dimeric enzymes in solution is substantially reduced, and a tetrahedral adduct of a substrate analogue is observed to be trapped at the active site in the crystal form. Remarkably, heating the dimeric enzymes increases activity. We propose that the homotetrameric structure of DHDPS reduces dynamic fluctuations present in the dimeric forms and increases specificity for the first substrate, pyruvate. By restricting motion in a key catalytic motif, a competing, non-productive reaction with a substrate analogue is avoided. Small-angle X-ray scattering and mutagenesis data, together with a B-factor analysis of the crystal structures, support this hypothesis and lead to the suggestion that in at least some cases, the evolution of quaternary enzyme structures might serve to optimise the dynamic properties of the protein subunits. © 2008 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Griffin, M.D.W. and Dobson, R.C.J. and Pearce, F.G. and Antonio, L. and Whitten, A.E. and Liew, C.K. and Mackay, J.P. and Trewhella, J. and Jameson, G.B. and Perugini, M.A. and Perugini, M.A. and Gerrard, J.A.\",\"doi\":\"10.1016/j.jmb.2008.05.038\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {Evolution of Quaternary Structure in a Homotetrameric Enzyme},\\n type = {article},\\n year = {2008},\\n pages = {691-703},\\n volume = {380},\\n id = {63b6023f-2683-3fd8-a96d-6ee13c0b2a51},\\n created = {2023-01-10T01:45:37.717Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:37.717Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Dihydrodipicolinate synthase (DHDPS) is an essential enzyme in (S)-lysine biosynthesis and an important antibiotic target. All X-ray crystal structures solved to date reveal a homotetrameric enzyme. In order to explore the role of this quaternary structure, dimeric variants of Escherichia coli DHDPS were engineered and their properties were compared to those of the wild-type tetrameric form. X-ray crystallography reveals that the active site is not disturbed when the quaternary structure is disrupted. However, the activity of the dimeric enzymes in solution is substantially reduced, and a tetrahedral adduct of a substrate analogue is observed to be trapped at the active site in the crystal form. Remarkably, heating the dimeric enzymes increases activity. We propose that the homotetrameric structure of DHDPS reduces dynamic fluctuations present in the dimeric forms and increases specificity for the first substrate, pyruvate. By restricting motion in a key catalytic motif, a competing, non-productive reaction with a substrate analogue is avoided. Small-angle X-ray scattering and mutagenesis data, together with a B-factor analysis of the crystal structures, support this hypothesis and lead to the suggestion that in at least some cases, the evolution of quaternary enzyme structures might serve to optimise the dynamic properties of the protein subunits. © 2008 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Griffin, M.D.W. and Dobson, R.C.J. and Pearce, F.G. and Antonio, L. and Whitten, A.E. and Liew, C.K. and Mackay, J.P. and Trewhella, J. and Jameson, G.B. and Perugini, M.A. and Perugini, M.A. and Gerrard, J.A.},\\n doi = {10.1016/j.jmb.2008.05.038},\\n journal = {Journal of Molecular Biology},\\n number = {4}\\n}\",\"author_short\":[\"Griffin,  M.\",\"Dobson,  R.\",\"Pearce,  F.\",\"Antonio,  L.\",\"Whitten,  A.\",\"Liew,  C.\",\"Mackay,  J.\",\"Trewhella,  J.\",\"Jameson,  G.\",\"Perugini,  M.\",\"Perugini,  M.\",\"Gerrard,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"griffin-dobson-pearce-antonio-whitten-liew-mackay-trewhella-etal-evolutionofquaternarystructureinahomotetramericenzyme-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Implementing the LIM code: The structural basis for cell type-specific assembly of LIM-homeodomain complexes\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"2018-2029\",\"volume\":\"27\",\"id\":\"aab1c421-98b0-388e-a789-d78011488b4f\",\"created\":\"2023-01-10T01:45:38.632Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:38.632Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"LIM-homeodomain (LIM-HD) transcription factors form a combinatorial 'LIM code' that contributes to the specification of cell types. In the ventral spinal cord, the binary LIM homeobox protein 3 (Lhx3)/LIM domain-binding protein 1 (Ldb1) complex specifies the formation of V2 interneurons. The additional expression of islet-1 (Isl1) in adjacent cells instead specifies the formation of motor neurons through assembly of a ternary complex in which Isl1 contacts both Lhx3 and Ldb1, displacing Lhx3 as the binding partner of Ldb1. However, little is known about how this molecular switch occurs. Here, we have identified the 30-residue Lhx3-binding domain on Isl1 (Isl1LBD). Although the LIM interaction domain of Ldb1 (Ldb1LID) and Isl1LBD share low levels of sequence homology, X-ray and NMR structures reveal that they bind Lhx3 in an identical manner, that is, Isl1LBD mimics Ldb1 LID. These data provide a structural basis for the formation of cell type-specific protein-protein interactions in which unstructured linear motifs with diverse sequences compete to bind protein partners. The resulting alternate protein complexes can target different genes to regulate key biological events. ©2008 European Molecular Biology Organization.\",\"bibtype\":\"article\",\"author\":\"Bhati, M. and Lee, C. and Nancarrow, A.L. and Lee, M. and Craig, V.J. and Bach, I. and Guss, J.M. and Mackay, J.P. and Matthews, J.M.\",\"doi\":\"10.1038/emboj.2008.123\",\"journal\":\"EMBO Journal\",\"number\":\"14\",\"bibtex\":\"@article{\\n title = {Implementing the LIM code: The structural basis for cell type-specific assembly of LIM-homeodomain complexes},\\n type = {article},\\n year = {2008},\\n pages = {2018-2029},\\n volume = {27},\\n id = {aab1c421-98b0-388e-a789-d78011488b4f},\\n created = {2023-01-10T01:45:38.632Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:38.632Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {LIM-homeodomain (LIM-HD) transcription factors form a combinatorial 'LIM code' that contributes to the specification of cell types. In the ventral spinal cord, the binary LIM homeobox protein 3 (Lhx3)/LIM domain-binding protein 1 (Ldb1) complex specifies the formation of V2 interneurons. The additional expression of islet-1 (Isl1) in adjacent cells instead specifies the formation of motor neurons through assembly of a ternary complex in which Isl1 contacts both Lhx3 and Ldb1, displacing Lhx3 as the binding partner of Ldb1. However, little is known about how this molecular switch occurs. Here, we have identified the 30-residue Lhx3-binding domain on Isl1 (Isl1LBD). Although the LIM interaction domain of Ldb1 (Ldb1LID) and Isl1LBD share low levels of sequence homology, X-ray and NMR structures reveal that they bind Lhx3 in an identical manner, that is, Isl1LBD mimics Ldb1 LID. These data provide a structural basis for the formation of cell type-specific protein-protein interactions in which unstructured linear motifs with diverse sequences compete to bind protein partners. The resulting alternate protein complexes can target different genes to regulate key biological events. ©2008 European Molecular Biology Organization.},\\n bibtype = {article},\\n author = {Bhati, M. and Lee, C. and Nancarrow, A.L. and Lee, M. and Craig, V.J. and Bach, I. and Guss, J.M. and Mackay, J.P. and Matthews, J.M.},\\n doi = {10.1038/emboj.2008.123},\\n journal = {EMBO Journal},\\n number = {14}\\n}\",\"author_short\":[\"Bhati,  M.\",\"Lee,  C.\",\"Nancarrow,  A.\",\"Lee,  M.\",\"Craig,  V.\",\"Bach,  I.\",\"Guss,  J.\",\"Mackay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"bhati-lee-nancarrow-lee-craig-bach-guss-mackay-etal-implementingthelimcodethestructuralbasisforcelltypespecificassemblyoflimhomeodomaincomplexes-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Structural and biophysical analysis of the DNA binding properties of myelin transcription factor 1\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"5158-5167\",\"volume\":\"283\",\"id\":\"915fd038-e9b1-36a4-8be4-dcecb9f56909\",\"created\":\"2023-01-10T01:45:39.551Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:39.551Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Zinc binding domains, or zinc fingers (ZnFs), form one of the most numerous and most diverse superclasses of protein structural motifs in eukaryotes. Although our understanding of the functions of several classes of these domains is relatively well developed, we know much less about the molecular mechanisms of action of many others. Myelin transcription factor 1 (MyT1) type ZnFs are found in organisms as diverse as nematodes and mammals and are found in a range of sequence contexts. MyT1, one of the early transcription factors expressed in the developing central nervous system, contains seven MyT1 ZnFs that are very highly conserved both within the protein and between species. We have used a range of biophysical techniques, including NMR spectroscopy and data-driven macromolecular docking, to investigate the structural basis for the interaction between MyT1 ZnFs and DNA. Our data indicate that MyT1 ZnFs recognize the major groove of DNA in a way that appears to differ from other known zinc binding domains. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Gamsjaeger, R. and Swanton, M.K. and Kobus, F.J. and Lehtomaki, E. and Lowry, J.A. and Kwan, A.H. and Matthews, J.M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M703772200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {Structural and biophysical analysis of the DNA binding properties of myelin transcription factor 1},\\n type = {article},\\n year = {2008},\\n pages = {5158-5167},\\n volume = {283},\\n id = {915fd038-e9b1-36a4-8be4-dcecb9f56909},\\n created = {2023-01-10T01:45:39.551Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:39.551Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Zinc binding domains, or zinc fingers (ZnFs), form one of the most numerous and most diverse superclasses of protein structural motifs in eukaryotes. Although our understanding of the functions of several classes of these domains is relatively well developed, we know much less about the molecular mechanisms of action of many others. Myelin transcription factor 1 (MyT1) type ZnFs are found in organisms as diverse as nematodes and mammals and are found in a range of sequence contexts. MyT1, one of the early transcription factors expressed in the developing central nervous system, contains seven MyT1 ZnFs that are very highly conserved both within the protein and between species. We have used a range of biophysical techniques, including NMR spectroscopy and data-driven macromolecular docking, to investigate the structural basis for the interaction between MyT1 ZnFs and DNA. Our data indicate that MyT1 ZnFs recognize the major groove of DNA in a way that appears to differ from other known zinc binding domains. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Gamsjaeger, R. and Swanton, M.K. and Kobus, F.J. and Lehtomaki, E. and Lowry, J.A. and Kwan, A.H. and Matthews, J.M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M703772200},\\n journal = {Journal of Biological Chemistry},\\n number = {8}\\n}\",\"author_short\":[\"Gamsjaeger,  R.\",\"Swanton,  M.\",\"Kobus,  F.\",\"Lehtomaki,  E.\",\"Lowry,  J.\",\"Kwan,  A.\",\"Matthews,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gamsjaeger-swanton-kobus-lehtomaki-lowry-kwan-matthews-mackay-structuralandbiophysicalanalysisofthednabindingpropertiesofmyelintranscriptionfactor1-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structure and inhibition of orotidine 5′-monophosphate decarboxylase from Plasmodium falciparum\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"3842-3854\",\"volume\":\"47\",\"id\":\"a141dd55-ab8c-3d60-ae66-5bb20bb77e87\",\"created\":\"2023-01-10T01:45:40.465Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:40.465Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Orotidine 5′-monophosphate (OMP) decarboxylase from Plasmodium falciparum (PfODCase, EC 4.1.1.23) has been overexpressed, purified, subjected to kinetic and biochemical analysis, and crystallized. The native enzyme is a homodimer with a subunit molecular mass of 38 kDa. The saturation curve for OMP as a substrate conformed to Michaelis-Menten kinetics with Km = 350 ± 60 nM and Vmax = 2.70 ± 0.10 μmol/min/mg protein. Inhibition patterns for nucleoside 5′-monophosphate analogues were linear competitive with respect to OMP with a decreasing potency of inhibition of PfODCase in the order: pyrazofurin 5′-monophosphate (Ki = 3.6 ± 0.7 nM) > xanthosine 5′-monophosphate (XMP, Ki = 4.4 ± 0.7 nM) > 6-azauridine 5′-monophosphate (AzaUMP, K i = 12 ±3 nM) > allopurinol-3-riboside 5′- monophosphate (Ki = 240 ± 20 nM). XMP is an ∼150-fold more potent inhibitor of PfODCase compared with the human enzyme. The structure of PfODCase was solved in the absence of ligand and displays a classic TIM-barrel fold characteristic of the enzyme. Both the phosphate-binding loop and the βα5-loop have conformational flexibility, which may be associated with substrate capture and product release along the reaction pathway. © 2008 American Chemical Society.\",\"bibtype\":\"article\",\"author\":\"Langley, D.B. and Shojaei, M. and Chan, C. and Hiu, C.L. and Mackay, J.P. and Traut, T.W. and Guss, J.M. and Christopherson, R.I.\",\"doi\":\"10.1021/bi702390k\",\"journal\":\"Biochemistry\",\"number\":\"12\",\"bibtex\":\"@article{\\n title = {Structure and inhibition of orotidine 5′-monophosphate decarboxylase from Plasmodium falciparum},\\n type = {article},\\n year = {2008},\\n pages = {3842-3854},\\n volume = {47},\\n id = {a141dd55-ab8c-3d60-ae66-5bb20bb77e87},\\n created = {2023-01-10T01:45:40.465Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:40.465Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Orotidine 5′-monophosphate (OMP) decarboxylase from Plasmodium falciparum (PfODCase, EC 4.1.1.23) has been overexpressed, purified, subjected to kinetic and biochemical analysis, and crystallized. The native enzyme is a homodimer with a subunit molecular mass of 38 kDa. The saturation curve for OMP as a substrate conformed to Michaelis-Menten kinetics with Km = 350 ± 60 nM and Vmax = 2.70 ± 0.10 μmol/min/mg protein. Inhibition patterns for nucleoside 5′-monophosphate analogues were linear competitive with respect to OMP with a decreasing potency of inhibition of PfODCase in the order: pyrazofurin 5′-monophosphate (Ki = 3.6 ± 0.7 nM) > xanthosine 5′-monophosphate (XMP, Ki = 4.4 ± 0.7 nM) > 6-azauridine 5′-monophosphate (AzaUMP, K i = 12 ±3 nM) > allopurinol-3-riboside 5′- monophosphate (Ki = 240 ± 20 nM). XMP is an ∼150-fold more potent inhibitor of PfODCase compared with the human enzyme. The structure of PfODCase was solved in the absence of ligand and displays a classic TIM-barrel fold characteristic of the enzyme. Both the phosphate-binding loop and the βα5-loop have conformational flexibility, which may be associated with substrate capture and product release along the reaction pathway. © 2008 American Chemical Society.},\\n bibtype = {article},\\n author = {Langley, D.B. and Shojaei, M. and Chan, C. and Hiu, C.L. and Mackay, J.P. and Traut, T.W. and Guss, J.M. and Christopherson, R.I.},\\n doi = {10.1021/bi702390k},\\n journal = {Biochemistry},\\n number = {12}\\n}\",\"author_short\":[\"Langley,  D.\",\"Shojaei,  M.\",\"Chan,  C.\",\"Hiu,  C.\",\"Mackay,  J.\",\"Traut,  T.\",\"Guss,  J.\",\"Christopherson,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"langley-shojaei-chan-hiu-mackay-traut-guss-christopherson-structureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparum-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Crystallization of a ZRANB2-RNA complex\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"1175-1177\",\"volume\":\"64\",\"id\":\"5c489013-8140-39aa-85b6-66fd1e85cec0\",\"created\":\"2023-01-10T01:45:41.371Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:41.371Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.\",\"bibtype\":\"article\",\"author\":\"Loughlin, F.E. and Lee, M. and Guss, J.M. and Mackay, J.P.\",\"doi\":\"10.1107/S1744309108036993\",\"journal\":\"Acta Crystallographica Section F: Structural Biology and Crystallization Communications\",\"number\":\"12\",\"bibtex\":\"@article{\\n title = {Crystallization of a ZRANB2-RNA complex},\\n type = {article},\\n year = {2008},\\n pages = {1175-1177},\\n volume = {64},\\n id = {5c489013-8140-39aa-85b6-66fd1e85cec0},\\n created = {2023-01-10T01:45:41.371Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:41.371Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.},\\n bibtype = {article},\\n author = {Loughlin, F.E. and Lee, M. and Guss, J.M. and Mackay, J.P.},\\n doi = {10.1107/S1744309108036993},\\n journal = {Acta Crystallographica Section F: Structural Biology and Crystallization Communications},\\n number = {12}\\n}\",\"author_short\":[\"Loughlin,  F.\",\"Lee,  M.\",\"Guss,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Designed metal-binding sites in biomolecular and bioinorganic interactions\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"484-490\",\"volume\":\"18\",\"id\":\"719ca520-9316-3d73-84b3-1e92163b1c32\",\"created\":\"2023-01-10T01:45:42.283Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:42.283Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The design of metal-binding functionality in proteins is expanding into many different areas with a wide range of practical and research applications. Here we review several developing areas of metal-related protein design, including the use of metals to induce protein-protein interactions or facilitate the assembly of extended nanostructures; the design of metallopeptides that bind metal and other inorganic surfaces, an area with potential in diverse applications ranging from nanoelectronics and photonics to biotechnology and biomedicine; and, the creation of sensitive and selective metal sensors for use both in vivo and in vitro. © 2008 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Matthews, J.M. and Loughlin, F.E. and Mackay, J.P.\",\"doi\":\"10.1016/j.sbi.2008.04.009\",\"journal\":\"Current Opinion in Structural Biology\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {Designed metal-binding sites in biomolecular and bioinorganic interactions},\\n type = {article},\\n year = {2008},\\n pages = {484-490},\\n volume = {18},\\n id = {719ca520-9316-3d73-84b3-1e92163b1c32},\\n created = {2023-01-10T01:45:42.283Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:42.283Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The design of metal-binding functionality in proteins is expanding into many different areas with a wide range of practical and research applications. Here we review several developing areas of metal-related protein design, including the use of metals to induce protein-protein interactions or facilitate the assembly of extended nanostructures; the design of metallopeptides that bind metal and other inorganic surfaces, an area with potential in diverse applications ranging from nanoelectronics and photonics to biotechnology and biomedicine; and, the creation of sensitive and selective metal sensors for use both in vivo and in vitro. © 2008 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Matthews, J.M. and Loughlin, F.E. and Mackay, J.P.},\\n doi = {10.1016/j.sbi.2008.04.009},\\n journal = {Current Opinion in Structural Biology},\\n number = {4}\\n}\",\"author_short\":[\"Matthews,  J.\",\"Loughlin,  F.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"matthews-loughlin-mackay-designedmetalbindingsitesinbiomolecularandbioinorganicinteractions-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Escherichia coli glucuronylsynthase: An engineered enzyme for the synthesis of β-glucuronides\",\"type\":\"article\",\"year\":\"2008\",\"pages\":\"1585-1588\",\"volume\":\"10\",\"id\":\"d014882a-497b-3d62-96a2-9f6d9e1e973d\",\"created\":\"2023-01-10T01:45:43.190Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:43.190Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The glycosynthase derived from E. coli β-glucuronidase catalyzes the glucuronylation of a range of primary, secondary, and aryl alcohols with moderate to excellent yields. The procedure provides an efficient, stereoselective, and scalable single-step synthesis of β-glucuronides under mild conditions. © 2008 American Chemical Society.\",\"bibtype\":\"article\",\"author\":\"Wilkinson, S.M. and Liew, C.W. and Mackay, J.P. and Salleh, H.M. and Withers, S.G. and McLeod, M.D.\",\"doi\":\"10.1021/ol8002767\",\"journal\":\"Organic Letters\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {Escherichia coli glucuronylsynthase: An engineered enzyme for the synthesis of β-glucuronides},\\n type = {article},\\n year = {2008},\\n pages = {1585-1588},\\n volume = {10},\\n id = {d014882a-497b-3d62-96a2-9f6d9e1e973d},\\n created = {2023-01-10T01:45:43.190Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:43.190Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The glycosynthase derived from E. coli β-glucuronidase catalyzes the glucuronylation of a range of primary, secondary, and aryl alcohols with moderate to excellent yields. The procedure provides an efficient, stereoselective, and scalable single-step synthesis of β-glucuronides under mild conditions. © 2008 American Chemical Society.},\\n bibtype = {article},\\n author = {Wilkinson, S.M. and Liew, C.W. and Mackay, J.P. and Salleh, H.M. and Withers, S.G. and McLeod, M.D.},\\n doi = {10.1021/ol8002767},\\n journal = {Organic Letters},\\n number = {8}\\n}\",\"author_short\":[\"Wilkinson,  S.\",\"Liew,  C.\",\"Mackay,  J.\",\"Salleh,  H.\",\"Withers,  S.\",\"McLeod,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wilkinson-liew-mackay-salleh-withers-mcleod-escherichiacoliglucuronylsynthaseanengineeredenzymeforthesynthesisofglucuronides-2008\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Grb7 SH2 domain structure and interactions with a cyclic peptide inhibitor of cancer cell migration and proliferation\",\"type\":\"article\",\"year\":\"2007\",\"volume\":\"7\",\"id\":\"4beae229-a038-3821-9cc9-5285dcb8776b\",\"created\":\"2023-01-10T01:45:44.136Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:44.136Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background. Human growth factor receptor bound protein 7 (Grb7) is an adapter protein that mediates the coupling of tyrosine kinases with their downstream signaling pathways. Grb7 is frequently overexpressed in invasive and metastatic human cancers and is implicated in cancer progression via its interaction with the ErbB2 receptor and focal adhesion kinase (FAK) that play critical roles in cell proliferation and migration. It is thus a prime target for the development of novel anti-cancer therapies. Recently, an inhibitory peptide (G7-18NATE) has been developed which binds specifically to the Grb7 SH2 domain and is able to attenuate cancer cell proliferation and migration in various cancer cell lines. Results. As a first step towards understanding how Grb7 may be inhibited by G7-18NATE, we solved the crystal structure of the Grb7 SH2 domain to 2.1 Å resolution. We describe the details of the peptide binding site underlying target specificity, as well as the dimer interface of Grb 7 SH2. Dimer formation of Grb7 was determined to be in the μM range using analytical ultracentrifugation for both full-length Grb7 and the SH2 domain alone, suggesting the SH2 domain forms the basis of a physiological dimer. ITC measurements of the interaction of the G7-18NATE peptide with the Grb7 SH2 domain revealed that it binds with a binding affinity of Kd= ∼35.7 μM and NMR spectroscopy titration experiments revealed that peptide binding causes perturbations to both the ligand binding surface of the Grb7 SH2 domain as well as to the dimer interface, suggesting that dimerisation of Grb7 is impacted on by peptide binding. Conclusion. Together the data allow us to propose a model of the Grb7 SH2 domain/G7-18NATE interaction and to rationalize the basis for the observed binding specificity and affinity. We propose that the current study will assist with the development of second generation Grb7 SH2 domain inhibitors, potentially leading to novel inhibitors of cancer cell migration and invasion. © 2007 Porter et al; licensee BioMed Central Ltd.\",\"bibtype\":\"article\",\"author\":\"Porter, C.J. and Matthews, J.M. and Mackay, J.P. and Pursglove, S.E. and Schmidberger, J.W. and Leedman, P.J. and Pero, S.C. and Krag, D.N. and Wilce, M.C.J. and Wilce, J.A.\",\"doi\":\"10.1186/1472-6807-7-58\",\"journal\":\"BMC Structural Biology\",\"bibtex\":\"@article{\\n title = {Grb7 SH2 domain structure and interactions with a cyclic peptide inhibitor of cancer cell migration and proliferation},\\n type = {article},\\n year = {2007},\\n volume = {7},\\n id = {4beae229-a038-3821-9cc9-5285dcb8776b},\\n created = {2023-01-10T01:45:44.136Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:44.136Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background. Human growth factor receptor bound protein 7 (Grb7) is an adapter protein that mediates the coupling of tyrosine kinases with their downstream signaling pathways. Grb7 is frequently overexpressed in invasive and metastatic human cancers and is implicated in cancer progression via its interaction with the ErbB2 receptor and focal adhesion kinase (FAK) that play critical roles in cell proliferation and migration. It is thus a prime target for the development of novel anti-cancer therapies. Recently, an inhibitory peptide (G7-18NATE) has been developed which binds specifically to the Grb7 SH2 domain and is able to attenuate cancer cell proliferation and migration in various cancer cell lines. Results. As a first step towards understanding how Grb7 may be inhibited by G7-18NATE, we solved the crystal structure of the Grb7 SH2 domain to 2.1 Å resolution. We describe the details of the peptide binding site underlying target specificity, as well as the dimer interface of Grb 7 SH2. Dimer formation of Grb7 was determined to be in the μM range using analytical ultracentrifugation for both full-length Grb7 and the SH2 domain alone, suggesting the SH2 domain forms the basis of a physiological dimer. ITC measurements of the interaction of the G7-18NATE peptide with the Grb7 SH2 domain revealed that it binds with a binding affinity of Kd= ∼35.7 μM and NMR spectroscopy titration experiments revealed that peptide binding causes perturbations to both the ligand binding surface of the Grb7 SH2 domain as well as to the dimer interface, suggesting that dimerisation of Grb7 is impacted on by peptide binding. Conclusion. Together the data allow us to propose a model of the Grb7 SH2 domain/G7-18NATE interaction and to rationalize the basis for the observed binding specificity and affinity. We propose that the current study will assist with the development of second generation Grb7 SH2 domain inhibitors, potentially leading to novel inhibitors of cancer cell migration and invasion. © 2007 Porter et al; licensee BioMed Central Ltd.},\\n bibtype = {article},\\n author = {Porter, C.J. and Matthews, J.M. and Mackay, J.P. and Pursglove, S.E. and Schmidberger, J.W. and Leedman, P.J. and Pero, S.C. and Krag, D.N. and Wilce, M.C.J. and Wilce, J.A.},\\n doi = {10.1186/1472-6807-7-58},\\n journal = {BMC Structural Biology}\\n}\",\"author_short\":[\"Porter,  C.\",\"Matthews,  J.\",\"Mackay,  J.\",\"Pursglove,  S.\",\"Schmidberger,  J.\",\"Leedman,  P.\",\"Pero,  S.\",\"Krag,  D.\",\"Wilce,  M.\",\"Wilce,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"porter-matthews-mackay-pursglove-schmidberger-leedman-pero-krag-etal-grb7sh2domainstructureandinteractionswithacyclicpeptideinhibitorofcancercellmigrationandproliferation-2007\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Sticky fingers: zinc-fingers as protein-recognition motifs\",\"type\":\"article\",\"year\":\"2007\",\"pages\":\"63-70\",\"volume\":\"32\",\"id\":\"0bdbbb8b-8f2a-39d3-a9c6-8cef76dd27e8\",\"created\":\"2023-01-10T01:45:45.060Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:45.060Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Zinc-fingers (ZnFs) are extremely abundant in higher eukaryotes. Once considered to function exclusively as sequence-specific DNA-binding motifs, ZnFs are now known to have additional activities such as the recognition of RNA and other proteins. Here we discuss recent advances in our understanding of ZnFs as specific modules for protein recognition. Structural studies of ZnF complexes reveal considerable diversity in terms of protein partners, binding modes and affinities, and highlight the often underestimated versatility of ZnF structure and function. An appreciation of the structural features of ZnF-protein interactions will contribute to our ability to engineer and to use ZnFs with tailored protein-binding properties. © 2006 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Gamsjaeger, R. and Liew, C.K. and Loughlin, F.E. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1016/j.tibs.2006.12.007\",\"journal\":\"Trends in Biochemical Sciences\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Sticky fingers: zinc-fingers as protein-recognition motifs},\\n type = {article},\\n year = {2007},\\n pages = {63-70},\\n volume = {32},\\n id = {0bdbbb8b-8f2a-39d3-a9c6-8cef76dd27e8},\\n created = {2023-01-10T01:45:45.060Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:45.060Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Zinc-fingers (ZnFs) are extremely abundant in higher eukaryotes. Once considered to function exclusively as sequence-specific DNA-binding motifs, ZnFs are now known to have additional activities such as the recognition of RNA and other proteins. Here we discuss recent advances in our understanding of ZnFs as specific modules for protein recognition. Structural studies of ZnF complexes reveal considerable diversity in terms of protein partners, binding modes and affinities, and highlight the often underestimated versatility of ZnF structure and function. An appreciation of the structural features of ZnF-protein interactions will contribute to our ability to engineer and to use ZnFs with tailored protein-binding properties. © 2006 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Gamsjaeger, R. and Liew, C.K. and Loughlin, F.E. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1016/j.tibs.2006.12.007},\\n journal = {Trends in Biochemical Sciences},\\n number = {2}\\n}\",\"author_short\":[\"Gamsjaeger,  R.\",\"Liew,  C.\",\"Loughlin,  F.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gamsjaeger-liew-loughlin-crossley-mackay-stickyfingerszincfingersasproteinrecognitionmotifs-2007\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution Structure of the THAP Domain from Caenorhabditis elegans C-terminal Binding Protein (CtBP)\",\"type\":\"article\",\"year\":\"2007\",\"pages\":\"382-390\",\"volume\":\"366\",\"id\":\"5e95220d-3a32-3190-836d-ac33617c7af2\",\"created\":\"2023-01-10T01:45:45.962Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:45.962Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The THAP (Thanatos-associated protein) domain is a recently discovered zinc-binding domain found in proteins involved in transcriptional regulation, cell-cycle control, apoptosis and chromatin modification. It contains a single zinc atom ligated by cysteine and histidine residues within a Cys-X2-4-Cys-X35-53-Cys-X2-His consensus. We have determined the NMR solution structure of the THAP domain from Caenorhabditis elegans C-terminal binding protein (CtBP) and show that it adopts a fold containing a treble clef motif, bearing similarity to the zinc finger-associated domain (ZAD) from Drosophila Grauzone. The CtBP THAP domain contains a large, positively charged surface patch and we demonstrate that this domain can bind to double-stranded DNA in an electrophoretic mobility-shift assay. These data, together with existing reports, indicate that THAP domains might exhibit a functional diversity similar to that observed for classical and GATA-type zinc fingers. © 2006 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Liew, C.K. and Crossley, M. and Mackay, J.P. and Nicholas, H.R.\",\"doi\":\"10.1016/j.jmb.2006.11.058\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Solution Structure of the THAP Domain from Caenorhabditis elegans C-terminal Binding Protein (CtBP)},\\n type = {article},\\n year = {2007},\\n pages = {382-390},\\n volume = {366},\\n id = {5e95220d-3a32-3190-836d-ac33617c7af2},\\n created = {2023-01-10T01:45:45.962Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:45.962Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The THAP (Thanatos-associated protein) domain is a recently discovered zinc-binding domain found in proteins involved in transcriptional regulation, cell-cycle control, apoptosis and chromatin modification. It contains a single zinc atom ligated by cysteine and histidine residues within a Cys-X2-4-Cys-X35-53-Cys-X2-His consensus. We have determined the NMR solution structure of the THAP domain from Caenorhabditis elegans C-terminal binding protein (CtBP) and show that it adopts a fold containing a treble clef motif, bearing similarity to the zinc finger-associated domain (ZAD) from Drosophila Grauzone. The CtBP THAP domain contains a large, positively charged surface patch and we demonstrate that this domain can bind to double-stranded DNA in an electrophoretic mobility-shift assay. These data, together with existing reports, indicate that THAP domains might exhibit a functional diversity similar to that observed for classical and GATA-type zinc fingers. © 2006 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Liew, C.K. and Crossley, M. and Mackay, J.P. and Nicholas, H.R.},\\n doi = {10.1016/j.jmb.2006.11.058},\\n journal = {Journal of Molecular Biology},\\n number = {2}\\n}\",\"author_short\":[\"Liew,  C.\",\"Crossley,  M.\",\"Mackay,  J.\",\"Nicholas,  H.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"liew-crossley-mackay-nicholas-solutionstructureofthethapdomainfromcaenorhabditiseleganscterminalbindingproteinctbp-2007\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"An erythroid chaperone that facilitates folding of α-globin subunits for hemoglobin synthesis\",\"type\":\"article\",\"year\":\"2007\",\"pages\":\"1856-1865\",\"volume\":\"117\",\"id\":\"b36c185f-d2dd-356f-b1e4-9fa2eef4f751\",\"created\":\"2023-01-10T01:45:46.881Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:46.881Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Erythrocyte precursors produce abundant α- and β-globin proteins, which assemble with each other to form hemoglobin A (HbA), the major blood oxygen carrier. αHb-stabilizing protein (AHSP) binds free α subunits reversibly to maintain their structure and limit their ability to generate reactive oxygen species. Accordingly, loss of AHSP aggravates the toxicity of excessive free α-globin caused by β-globin gene disruption in mice. Surprisingly, we found that AHSP also has important functions when free α-globin is limited. Thus, compound mutants lacking both Ahsp and 1 of 4 α-globin genes (genotype Ahsp-/-α-globin -/-α/αα) exhibited more severe anemia and Hb instability than mice with either mutation alone. In vitro, recombinant AHSP promoted folding of newly translated α-globin, enhanced its refolding after denaturation, and facilitated its incorporation into HbA. Moreover, in erythroid precursors, newly formed free α-globin was destabilized by loss of AHSP. Therefore, in addition to its previously defined role in detoxification of excess α-globin, AHSP also acts as a molecular chaperone to stabilize nascent α-globin for HbA assembly. Our findings illustrate what we believe to be a novel adaptive mechanism by which a specialized cell coordinates high-level production of a multisubunit protein and protects against various synthetic imbalances.\",\"bibtype\":\"article\",\"author\":\"Yu, X. and Kong, Y. and Dore, L.C. and Abdulmalik, O. and Katein, A.M. and Zhou, S. and Choi, J.K. and Gell, D. and Mackay, J.P. and Gow, A.J. and Gow, A.J. and Weiss, M.J.\",\"doi\":\"10.1172/JCI31664\",\"journal\":\"Journal of Clinical Investigation\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {An erythroid chaperone that facilitates folding of α-globin subunits for hemoglobin synthesis},\\n type = {article},\\n year = {2007},\\n pages = {1856-1865},\\n volume = {117},\\n id = {b36c185f-d2dd-356f-b1e4-9fa2eef4f751},\\n created = {2023-01-10T01:45:46.881Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:46.881Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Erythrocyte precursors produce abundant α- and β-globin proteins, which assemble with each other to form hemoglobin A (HbA), the major blood oxygen carrier. αHb-stabilizing protein (AHSP) binds free α subunits reversibly to maintain their structure and limit their ability to generate reactive oxygen species. Accordingly, loss of AHSP aggravates the toxicity of excessive free α-globin caused by β-globin gene disruption in mice. Surprisingly, we found that AHSP also has important functions when free α-globin is limited. Thus, compound mutants lacking both Ahsp and 1 of 4 α-globin genes (genotype Ahsp-/-α-globin -/-α/αα) exhibited more severe anemia and Hb instability than mice with either mutation alone. In vitro, recombinant AHSP promoted folding of newly translated α-globin, enhanced its refolding after denaturation, and facilitated its incorporation into HbA. Moreover, in erythroid precursors, newly formed free α-globin was destabilized by loss of AHSP. Therefore, in addition to its previously defined role in detoxification of excess α-globin, AHSP also acts as a molecular chaperone to stabilize nascent α-globin for HbA assembly. Our findings illustrate what we believe to be a novel adaptive mechanism by which a specialized cell coordinates high-level production of a multisubunit protein and protects against various synthetic imbalances.},\\n bibtype = {article},\\n author = {Yu, X. and Kong, Y. and Dore, L.C. and Abdulmalik, O. and Katein, A.M. and Zhou, S. and Choi, J.K. and Gell, D. and Mackay, J.P. and Gow, A.J. and Gow, A.J. and Weiss, M.J.},\\n doi = {10.1172/JCI31664},\\n journal = {Journal of Clinical Investigation},\\n number = {7}\\n}\",\"author_short\":[\"Yu,  X.\",\"Kong,  Y.\",\"Dore,  L.\",\"Abdulmalik,  O.\",\"Katein,  A.\",\"Zhou,  S.\",\"Choi,  J.\",\"Gell,  D.\",\"Mackay,  J.\",\"Gow,  A.\",\"Gow,  A.\",\"Weiss,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"yu-kong-dore-abdulmalik-katein-zhou-choi-gell-etal-anerythroidchaperonethatfacilitatesfoldingofglobinsubunitsforhemoglobinsynthesis-2007\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Mutations in crdiac T-box factor gene TBX20 are associated with diverse cardiac pathologies, including defects of septation and valvulogenesis and cardiomyopathy\",\"type\":\"article\",\"year\":\"2007\",\"pages\":\"280-291\",\"volume\":\"81\",\"id\":\"52c04b2f-8782-380e-959a-24222788591c\",\"created\":\"2023-01-10T01:45:47.818Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:47.818Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The T-box family transcription factor gene TBX20 acts in a conserved regulatory network, guiding heart formation and patterning in diverse species. Mouse Tbx20 is expressed in cardiac progenitor cells, differentiating cardiomyocytes, and developing valvular tissue, and its deletion or RNA interference-mediated knockdown is catastrophic for heart development. TBX20 interacts physically, functionally, and genetically with other cardiac transcription factors, including NKX2-5, GATA4, and TBX5, mutations of which cause congenital heart disease (CHD). Here, we report nonsense (Q195X) and missense (I152M) germline mutations within the T-box DNA-binding domain of human TBX20 that were associated with a family history of CHD and a complex spectrum of developmental anomalies, including defects in septation, chamber growth, and valvulogenesis. Biophysical characterization of wild-type and mutant proteins indicated how the missense mutation disrupts the structure and function of the TBX20 T-box. Dilated cardiomyopathy was a feature of the TBX20 mutant phenotype in humans and mice, suggesting that mutations in developmental transcription factors can provide a sensitized template for adult-onset heart disease. Our findings are the first to link TBX20 mutations to human pathology. They provide insights into how mutation of different genes in an interactive regulatory circuit lead to diverse clinical phenotypes, with implications for diagnosis, genetic screening, and patient follow-up. © 2007 by The American Society of Human Genetics. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Kirk, E.P. and Sunde, M. and Costa, M.W. and Rankin, S.A. and Wolstein, O. and Castro, M.L. and Butler, T.L. and Hyun, C. and Guo, G. and Otway, R. and Winlaw, D.S. and Harvey, R.P.\",\"doi\":\"10.1086/519530\",\"journal\":\"American Journal of Human Genetics\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Mutations in crdiac T-box factor gene TBX20 are associated with diverse cardiac pathologies, including defects of septation and valvulogenesis and cardiomyopathy},\\n type = {article},\\n year = {2007},\\n pages = {280-291},\\n volume = {81},\\n id = {52c04b2f-8782-380e-959a-24222788591c},\\n created = {2023-01-10T01:45:47.818Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:47.818Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The T-box family transcription factor gene TBX20 acts in a conserved regulatory network, guiding heart formation and patterning in diverse species. Mouse Tbx20 is expressed in cardiac progenitor cells, differentiating cardiomyocytes, and developing valvular tissue, and its deletion or RNA interference-mediated knockdown is catastrophic for heart development. TBX20 interacts physically, functionally, and genetically with other cardiac transcription factors, including NKX2-5, GATA4, and TBX5, mutations of which cause congenital heart disease (CHD). Here, we report nonsense (Q195X) and missense (I152M) germline mutations within the T-box DNA-binding domain of human TBX20 that were associated with a family history of CHD and a complex spectrum of developmental anomalies, including defects in septation, chamber growth, and valvulogenesis. Biophysical characterization of wild-type and mutant proteins indicated how the missense mutation disrupts the structure and function of the TBX20 T-box. Dilated cardiomyopathy was a feature of the TBX20 mutant phenotype in humans and mice, suggesting that mutations in developmental transcription factors can provide a sensitized template for adult-onset heart disease. Our findings are the first to link TBX20 mutations to human pathology. They provide insights into how mutation of different genes in an interactive regulatory circuit lead to diverse clinical phenotypes, with implications for diagnosis, genetic screening, and patient follow-up. © 2007 by The American Society of Human Genetics. All rights reserved.},\\n bibtype = {article},\\n author = {Kirk, E.P. and Sunde, M. and Costa, M.W. and Rankin, S.A. and Wolstein, O. and Castro, M.L. and Butler, T.L. and Hyun, C. and Guo, G. and Otway, R. and Winlaw, D.S. and Harvey, R.P.},\\n doi = {10.1086/519530},\\n journal = {American Journal of Human Genetics},\\n number = {2}\\n}\",\"author_short\":[\"Kirk,  E.\",\"Sunde,  M.\",\"Costa,  M.\",\"Rankin,  S.\",\"Wolstein,  O.\",\"Castro,  M.\",\"Butler,  T.\",\"Hyun,  C.\",\"Guo,  G.\",\"Otway,  R.\",\"Winlaw,  D.\",\"Harvey,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kirk-sunde-costa-rankin-wolstein-castro-butler-hyun-etal-mutationsincrdiactboxfactorgenetbx20areassociatedwithdiversecardiacpathologiesincludingdefectsofseptationandvalvulogenesisandcardiomyopathy-2007\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Protein interactions: is seeing believing?\",\"type\":\"article\",\"year\":\"2007\",\"pages\":\"530-531\",\"volume\":\"32\",\"id\":\"44b87550-332a-36df-aa40-16703ffad894\",\"created\":\"2023-01-10T01:45:48.739Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:48.739Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Sunde, M. and Lowry, J.A. and Crossley, M. and Matthews, J.M.\",\"doi\":\"10.1016/j.tibs.2007.09.006\",\"journal\":\"Trends in Biochemical Sciences\",\"number\":\"12\",\"bibtex\":\"@article{\\n title = {Protein interactions: is seeing believing?},\\n type = {article},\\n year = {2007},\\n pages = {530-531},\\n volume = {32},\\n id = {44b87550-332a-36df-aa40-16703ffad894},\\n created = {2023-01-10T01:45:48.739Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:48.739Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Mackay, J.P. and Sunde, M. and Lowry, J.A. and Crossley, M. and Matthews, J.M.},\\n doi = {10.1016/j.tibs.2007.09.006},\\n journal = {Trends in Biochemical Sciences},\\n number = {12}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Sunde,  M.\",\"Lowry,  J.\",\"Crossley,  M.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-sunde-lowry-crossley-matthews-proteininteractionsisseeingbelieving-2007\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Crystal structures of flax rust avirulence proteins AvrL567-A and -D reveal details of the structural basis for flax disease resistance specificity\",\"type\":\"article\",\"year\":\"2007\",\"pages\":\"2898-2912\",\"volume\":\"19\",\"id\":\"e42e696d-217e-3568-9613-ef60ec204073\",\"created\":\"2023-01-10T01:45:49.665Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:49.665Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The gene-for-gene mechanism of plant disease resistance involves direct or indirect recognition of pathogen avirulence (Avr) proteins by plant resistance (R) proteins. Flax rust (Melampsora lini) AvrL567 avirulence proteins and the corresponding flax (Linum usitatissimum) L5, L6, and L7 resistance proteins interact directly. We determined the three-dimensional structures of two members of the AvrL567 family, AvrL567-A and AvrL567-D, at 1.4- and 2.3-Å resolution, respectively. The structures of both proteins are very similar and reveal a β-sandwich fold with no close known structural homologs. The polymorphic residues in the AvrL567 family map to the surface of the protein, and polymorphisms in residues associated with recognition differences for the R proteins lead to significant changes in surface chemical properties. Analysis of single amino acid substitutions in AvrL567 proteins confirm the role of individual residues in conferring differences in recognition and suggest that the specificity results from the cumulative effects of multiple amino acid contacts. The structures also provide insights into possible pathogen-associated functions of AvrL567 proteins, with nucleic acid binding activity demonstrated in vitro. Our studies provide some of the first structural information on avirulence proteins that bind directly to the corresponding resistance proteins, allowing an examination of the molecular basis of the interaction with the resistance proteins as a step toward designing new resistance specificities. © 2007 American Society of Plant Biologists.\",\"bibtype\":\"article\",\"author\":\"Wang, C.-I.A. and Gunčar, G. and Forwood, J.K. and Teh, T. and Catanzariti, A.-M. and Lawrence, G.J. and Loughlin, F.E. and Mackay, J.P. and Schirra, H.J. and Anderson, P.A. and Dodds, P.N. and Kobe, B.\",\"doi\":\"10.1105/tpc.107.053611\",\"journal\":\"Plant Cell\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {Crystal structures of flax rust avirulence proteins AvrL567-A and -D reveal details of the structural basis for flax disease resistance specificity},\\n type = {article},\\n year = {2007},\\n pages = {2898-2912},\\n volume = {19},\\n id = {e42e696d-217e-3568-9613-ef60ec204073},\\n created = {2023-01-10T01:45:49.665Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:49.665Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The gene-for-gene mechanism of plant disease resistance involves direct or indirect recognition of pathogen avirulence (Avr) proteins by plant resistance (R) proteins. Flax rust (Melampsora lini) AvrL567 avirulence proteins and the corresponding flax (Linum usitatissimum) L5, L6, and L7 resistance proteins interact directly. We determined the three-dimensional structures of two members of the AvrL567 family, AvrL567-A and AvrL567-D, at 1.4- and 2.3-Å resolution, respectively. The structures of both proteins are very similar and reveal a β-sandwich fold with no close known structural homologs. The polymorphic residues in the AvrL567 family map to the surface of the protein, and polymorphisms in residues associated with recognition differences for the R proteins lead to significant changes in surface chemical properties. Analysis of single amino acid substitutions in AvrL567 proteins confirm the role of individual residues in conferring differences in recognition and suggest that the specificity results from the cumulative effects of multiple amino acid contacts. The structures also provide insights into possible pathogen-associated functions of AvrL567 proteins, with nucleic acid binding activity demonstrated in vitro. Our studies provide some of the first structural information on avirulence proteins that bind directly to the corresponding resistance proteins, allowing an examination of the molecular basis of the interaction with the resistance proteins as a step toward designing new resistance specificities. © 2007 American Society of Plant Biologists.},\\n bibtype = {article},\\n author = {Wang, C.-I.A. and Gunčar, G. and Forwood, J.K. and Teh, T. and Catanzariti, A.-M. and Lawrence, G.J. and Loughlin, F.E. and Mackay, J.P. and Schirra, H.J. and Anderson, P.A. and Dodds, P.N. and Kobe, B.},\\n doi = {10.1105/tpc.107.053611},\\n journal = {Plant Cell},\\n number = {9}\\n}\",\"author_short\":[\"Wang,  C.\",\"Gunčar,  G.\",\"Forwood,  J.\",\"Teh,  T.\",\"Catanzariti,  A.\",\"Lawrence,  G.\",\"Loughlin,  F.\",\"Mackay,  J.\",\"Schirra,  H.\",\"Anderson,  P.\",\"Dodds,  P.\",\"Kobe,  B.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wang-gunar-forwood-teh-catanzariti-lawrence-loughlin-mackay-etal-crystalstructuresofflaxrustavirulenceproteinsavrl567aanddrevealdetailsofthestructuralbasisforflaxdiseaseresistancespecificity-2007\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Zinc fingers are known as domains for binding DNA and RNA. Do they also mediate protein-protein interactions?\",\"type\":\"article\",\"year\":\"2006\",\"pages\":\"731-733\",\"volume\":\"58\",\"id\":\"a59a3ace-fadf-3a89-ab6c-424f0813e6d6\",\"created\":\"2023-01-10T01:45:50.577Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:50.577Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Loughlin, F.E. and Mackay, J.P.\",\"doi\":\"10.1080/15216540600868445\",\"journal\":\"IUBMB Life\",\"number\":\"12\",\"bibtex\":\"@article{\\n title = {Zinc fingers are known as domains for binding DNA and RNA. Do they also mediate protein-protein interactions?},\\n type = {article},\\n year = {2006},\\n pages = {731-733},\\n volume = {58},\\n id = {a59a3ace-fadf-3a89-ab6c-424f0813e6d6},\\n created = {2023-01-10T01:45:50.577Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:50.577Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Loughlin, F.E. and Mackay, J.P.},\\n doi = {10.1080/15216540600868445},\\n journal = {IUBMB Life},\\n number = {12}\\n}\",\"author_short\":[\"Loughlin,  F.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"loughlin-mackay-zincfingersareknownasdomainsforbindingdnaandrnadotheyalsomediateproteinproteininteractions-2006\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Molecular analysis of the interaction between the hematopoietic master transcription factors GATA-1 and PU\",\"type\":\"article\",\"year\":\"2006\",\"pages\":\"28296-28306\",\"volume\":\"281\",\"id\":\"62d88558-80bb-327f-9a17-f07a26210a6c\",\"created\":\"2023-01-10T01:45:51.530Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:51.530Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"GATA-1 and PU.1 are transcription factors that control erythroid and myeloid development, respectively. The two proteins have been shown to function in an antagonistic fashion, with GATA-1 repressing PU.1 activity during erythropoiesis and PU.1 repressing GATA-1 function during myelopoiesis. It has also become clear that this functional antagonism involves direct interactions between the two proteins. However, the molecular basis for these interactions is not known, and a number of inconsistencies exist in the literature. We have used a range of biophysical methods to define the molecular details of the GATA-1-PU.1 interaction. A combination of NMR titration data and extensive mutagenesis revealed that the PU.1-Ets domain and the GATA-1 C-terminal zinc finger (CF) form a low affinity interaction in which specific regions of each protein are implicated. Surprisingly, the interaction cannot be disrupted by single alanine substitution mutations, suggesting that binding is distributed over an extended interface. The C-terminal basic tail region of CF appears to be sufficient to mediate an interaction with PU.1-Ets, and neither acetylation nor phosphorylation of a peptide corresponding to this region disrupts binding, indicating that the interaction is not dominated by electrostatic interactions. The CF basic tail shares significant sequence homology with the PU.1 interacting motif from c-Jun, suggesting that GATA-1 and c-Jun might compete to bind PU.1. Taken together, our data provide a molecular perspective on the GATA-1-PU.1 interaction, resolving several issues in the existing data and providing insight into the mechanisms through which these two proteins combine to regulate blood development. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Chu, W.L. and Rand, K.D. and Simpson, R.J.Y. and Yung, W.W. and Mansfield, R.E. and Crossley, M. and Prœtorius-Ibba, M. and Nerlov, C. and Poulsen, F.M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M602830200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"38\",\"bibtex\":\"@article{\\n title = {Molecular analysis of the interaction between the hematopoietic master transcription factors GATA-1 and PU},\\n type = {article},\\n year = {2006},\\n pages = {28296-28306},\\n volume = {281},\\n id = {62d88558-80bb-327f-9a17-f07a26210a6c},\\n created = {2023-01-10T01:45:51.530Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:51.530Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {GATA-1 and PU.1 are transcription factors that control erythroid and myeloid development, respectively. The two proteins have been shown to function in an antagonistic fashion, with GATA-1 repressing PU.1 activity during erythropoiesis and PU.1 repressing GATA-1 function during myelopoiesis. It has also become clear that this functional antagonism involves direct interactions between the two proteins. However, the molecular basis for these interactions is not known, and a number of inconsistencies exist in the literature. We have used a range of biophysical methods to define the molecular details of the GATA-1-PU.1 interaction. A combination of NMR titration data and extensive mutagenesis revealed that the PU.1-Ets domain and the GATA-1 C-terminal zinc finger (CF) form a low affinity interaction in which specific regions of each protein are implicated. Surprisingly, the interaction cannot be disrupted by single alanine substitution mutations, suggesting that binding is distributed over an extended interface. The C-terminal basic tail region of CF appears to be sufficient to mediate an interaction with PU.1-Ets, and neither acetylation nor phosphorylation of a peptide corresponding to this region disrupts binding, indicating that the interaction is not dominated by electrostatic interactions. The CF basic tail shares significant sequence homology with the PU.1 interacting motif from c-Jun, suggesting that GATA-1 and c-Jun might compete to bind PU.1. Taken together, our data provide a molecular perspective on the GATA-1-PU.1 interaction, resolving several issues in the existing data and providing insight into the mechanisms through which these two proteins combine to regulate blood development. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Chu, W.L. and Rand, K.D. and Simpson, R.J.Y. and Yung, W.W. and Mansfield, R.E. and Crossley, M. and Prœtorius-Ibba, M. and Nerlov, C. and Poulsen, F.M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M602830200},\\n journal = {Journal of Biological Chemistry},\\n number = {38}\\n}\",\"author_short\":[\"Chu,  W.\",\"Rand,  K.\",\"Simpson,  R.\",\"Yung,  W.\",\"Mansfield,  R.\",\"Crossley,  M.\",\"Prœtorius-Ibba,  M.\",\"Nerlov,  C.\",\"Poulsen,  F.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"chu-rand-simpson-yung-mansfield-crossley-prtoriusibba-nerlov-etal-molecularanalysisoftheinteractionbetweenthehematopoieticmastertranscriptionfactorsgata1andpu-2006\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Identification of the Key LMO2-binding Determinants on Ldb1\",\"type\":\"article\",\"year\":\"2006\",\"pages\":\"66-75\",\"volume\":\"359\",\"id\":\"14513112-4f7f-308b-b9ed-cb4aa5fd5f5b\",\"created\":\"2023-01-10T01:45:52.515Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:52.515Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The overexpression of LIM-only protein 2 (LMO2) in T-cells, as a result of chromosomal translocations, retroviral insertion during gene therapy, or in transgenic mice models, leads to the onset of T-cell leukemias. LMO2 comprises two protein-binding LIM domains that allow LMO2 to interact with multiple protein partners, including LIM domain-binding protein 1 (Ldb1, also known as CLIM2 and NLI), an essential cofactor for LMO proteins. Sequestration of Ldb1 by LMO2 in T-cells may prevent it binding other key partners, such as LMO4. Here, we show using protein engineering and enzyme-linked immunosorbent assay (ELISA) methodologies that LMO2 binds Ldb1 with a twofold lower affinity than does LMO4. Thus, excess LMO2 rather than an intrinsically higher binding affinity would lead to sequestration of Ldb1. Both LIM domains of LMO2 are required for high-affinity binding to Ldb1 (KD=2.0×10-8 M). However, the first LIM domain of LMO2 is primarily responsible for binding to Ldb1 (KD=2.3×10-7 M), whereas the second LIM domain increases binding by an order of magnitude. We used mutagenesis in combination with yeast two-hybrid analysis, and phage display selection to identify LMO2-binding \\\"hot spots\\\" within Ldb1 that locate to the LIM1-binding region. The delineation of this region reveals some specific differences when compared to the equivalent LMO4:Ldb1 interaction that hold promise for the development of reagents to specifically bind LMO2 in the treatment of leukemia. © 2006 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Ryan, D.P. and Sunde, M. and Kwan, A.H.-Y. and Marianayagam, N.J. and Nancarrow, A.L. and vanden Hoven, R.N. and Thompson, L.S. and Baca, M. and Mackay, J.P. and Visvader, J.E. and Visvader, J.E. and Matthews, J.M.\",\"doi\":\"10.1016/j.jmb.2006.02.074\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Identification of the Key LMO2-binding Determinants on Ldb1},\\n type = {article},\\n year = {2006},\\n pages = {66-75},\\n volume = {359},\\n id = {14513112-4f7f-308b-b9ed-cb4aa5fd5f5b},\\n created = {2023-01-10T01:45:52.515Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:52.515Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The overexpression of LIM-only protein 2 (LMO2) in T-cells, as a result of chromosomal translocations, retroviral insertion during gene therapy, or in transgenic mice models, leads to the onset of T-cell leukemias. LMO2 comprises two protein-binding LIM domains that allow LMO2 to interact with multiple protein partners, including LIM domain-binding protein 1 (Ldb1, also known as CLIM2 and NLI), an essential cofactor for LMO proteins. Sequestration of Ldb1 by LMO2 in T-cells may prevent it binding other key partners, such as LMO4. Here, we show using protein engineering and enzyme-linked immunosorbent assay (ELISA) methodologies that LMO2 binds Ldb1 with a twofold lower affinity than does LMO4. Thus, excess LMO2 rather than an intrinsically higher binding affinity would lead to sequestration of Ldb1. Both LIM domains of LMO2 are required for high-affinity binding to Ldb1 (KD=2.0×10-8 M). However, the first LIM domain of LMO2 is primarily responsible for binding to Ldb1 (KD=2.3×10-7 M), whereas the second LIM domain increases binding by an order of magnitude. We used mutagenesis in combination with yeast two-hybrid analysis, and phage display selection to identify LMO2-binding \\\"hot spots\\\" within Ldb1 that locate to the LIM1-binding region. The delineation of this region reveals some specific differences when compared to the equivalent LMO4:Ldb1 interaction that hold promise for the development of reagents to specifically bind LMO2 in the treatment of leukemia. © 2006 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Ryan, D.P. and Sunde, M. and Kwan, A.H.-Y. and Marianayagam, N.J. and Nancarrow, A.L. and vanden Hoven, R.N. and Thompson, L.S. and Baca, M. and Mackay, J.P. and Visvader, J.E. and Visvader, J.E. and Matthews, J.M.},\\n doi = {10.1016/j.jmb.2006.02.074},\\n journal = {Journal of Molecular Biology},\\n number = {1}\\n}\",\"author_short\":[\"Ryan,  D.\",\"Sunde,  M.\",\"Kwan,  A.\",\"Marianayagam,  N.\",\"Nancarrow,  A.\",\"vanden Hoven,  R.\",\"Thompson,  L.\",\"Baca,  M.\",\"Mackay,  J.\",\"Visvader,  J.\",\"Visvader,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"ryan-sunde-kwan-marianayagam-nancarrow-vandenhoven-thompson-baca-etal-identificationofthekeylmo2bindingdeterminantsonldb1-2006\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural basis for rodlet assembly in fungal hydrophobins\",\"type\":\"article\",\"year\":\"2006\",\"pages\":\"3621-3626\",\"volume\":\"103\",\"id\":\"0944c3ce-33ab-33ae-b13f-43193a7e6f03\",\"created\":\"2023-01-10T01:45:53.429Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:53.429Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.\",\"bibtype\":\"article\",\"author\":\"Kwan, A.H.Y. and Winefield, R.D. and Sunde, M. and Matthews, J.M. and Haverkamp, R.G. and Templeton, M.D. and Mackay, J.P.\",\"doi\":\"10.1073/pnas.0505704103\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {Structural basis for rodlet assembly in fungal hydrophobins},\\n type = {article},\\n year = {2006},\\n pages = {3621-3626},\\n volume = {103},\\n id = {0944c3ce-33ab-33ae-b13f-43193a7e6f03},\\n created = {2023-01-10T01:45:53.429Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:53.429Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.},\\n bibtype = {article},\\n author = {Kwan, A.H.Y. and Winefield, R.D. and Sunde, M. and Matthews, J.M. and Haverkamp, R.G. and Templeton, M.D. and Mackay, J.P.},\\n doi = {10.1073/pnas.0505704103},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {10}\\n}\",\"author_short\":[\"Kwan,  A.\",\"Winefield,  R.\",\"Sunde,  M.\",\"Matthews,  J.\",\"Haverkamp,  R.\",\"Templeton,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Analysis of the structure and function of the transcriptional coregulator HOP\",\"type\":\"article\",\"year\":\"2006\",\"pages\":\"10584-10590\",\"volume\":\"45\",\"id\":\"ad62d749-5828-3220-9e44-df8440515254\",\"created\":\"2023-01-10T01:45:54.347Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:54.347Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Homeodomain-only protein (HOP) is an 8-kDa transcriptional corepressor that is essential for the normal development of the mammalian heart. Previous studies have shown that HOP, which consists entirely of a putative homeodomain, acts downstream of Nkx2.5 and associates with the serum response factor (SRF), repressing transcription from SRF-responsive genes. HOP is also able to recruit histone deacetylase (HDAC) activity, consistent with its ability to repress transcription. Unlike other classic homeodomain proteins, HOP does not appear to interact with DNA, although it has been unclear if this is because of an overall divergent structure or because of specific amino acid differences between HOP and other homeodomains. To work toward an understanding of HOP function, we have determined the 3D structure of full-length HOP and used a range of biochemical assays to define the parts of the protein that are functionally important for its repression activity. We show that HOP forms a classical homeodomain fold but that it cannot recognize double stranded DNA, a result that emphasizes the importance of caution in predicting protein function from sequence homology alone. We also demonstrate that two distinct regions on the surface of HOP are required for its ability to repress an SRF-driven reporter gene, and it is likely that these motifs direct interactions between HOP and partner proteins such as SRF- and HDAC-containing complexes. Our results demonstrate that the homeodomain fold has been co-opted during evolution for functions other than sequence-specific DNA binding and suggest that HOP functions as an adaptor protein to mediate transcriptional repression. © 2006 American Chemical Society.\",\"bibtype\":\"article\",\"author\":\"Kook, H. and Yung, W.W. and Simpson, R.J. and Kee, H.J. and Shin, S. and Lowry, J.A. and Loughlin, F.E. and Yin, Z. and Epstein, J.A. and Mackay, J.P.\",\"doi\":\"10.1021/bi060641s\",\"journal\":\"Biochemistry\",\"number\":\"35\",\"bibtex\":\"@article{\\n title = {Analysis of the structure and function of the transcriptional coregulator HOP},\\n type = {article},\\n year = {2006},\\n pages = {10584-10590},\\n volume = {45},\\n id = {ad62d749-5828-3220-9e44-df8440515254},\\n created = {2023-01-10T01:45:54.347Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:54.347Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Homeodomain-only protein (HOP) is an 8-kDa transcriptional corepressor that is essential for the normal development of the mammalian heart. Previous studies have shown that HOP, which consists entirely of a putative homeodomain, acts downstream of Nkx2.5 and associates with the serum response factor (SRF), repressing transcription from SRF-responsive genes. HOP is also able to recruit histone deacetylase (HDAC) activity, consistent with its ability to repress transcription. Unlike other classic homeodomain proteins, HOP does not appear to interact with DNA, although it has been unclear if this is because of an overall divergent structure or because of specific amino acid differences between HOP and other homeodomains. To work toward an understanding of HOP function, we have determined the 3D structure of full-length HOP and used a range of biochemical assays to define the parts of the protein that are functionally important for its repression activity. We show that HOP forms a classical homeodomain fold but that it cannot recognize double stranded DNA, a result that emphasizes the importance of caution in predicting protein function from sequence homology alone. We also demonstrate that two distinct regions on the surface of HOP are required for its ability to repress an SRF-driven reporter gene, and it is likely that these motifs direct interactions between HOP and partner proteins such as SRF- and HDAC-containing complexes. Our results demonstrate that the homeodomain fold has been co-opted during evolution for functions other than sequence-specific DNA binding and suggest that HOP functions as an adaptor protein to mediate transcriptional repression. © 2006 American Chemical Society.},\\n bibtype = {article},\\n author = {Kook, H. and Yung, W.W. and Simpson, R.J. and Kee, H.J. and Shin, S. and Lowry, J.A. and Loughlin, F.E. and Yin, Z. and Epstein, J.A. and Mackay, J.P.},\\n doi = {10.1021/bi060641s},\\n journal = {Biochemistry},\\n number = {35}\\n}\",\"author_short\":[\"Kook,  H.\",\"Yung,  W.\",\"Simpson,  R.\",\"Kee,  H.\",\"Shin,  S.\",\"Lowry,  J.\",\"Loughlin,  F.\",\"Yin,  Z.\",\"Epstein,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kook-yung-simpson-kee-shin-lowry-loughlin-yin-etal-analysisofthestructureandfunctionofthetranscriptionalcoregulatorhop-2006\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP\",\"type\":\"article\",\"year\":\"2006\",\"pages\":\"287-297\",\"volume\":\"362\",\"id\":\"f66fd98b-e46d-356c-adc5-dbb7bb45a1c1\",\"created\":\"2023-01-10T01:45:55.259Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:55.259Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Gell, D.A. and Westman, B.J. and Gorman, D. and Liew, C. and Welch, J.J. and Weiss, M.J. and Mackay, J.P.\",\"doi\":\"10.1016/j.jmb.2006.07.010\",\"journal\":\"Journal of Molecular Biology\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP},\\n type = {article},\\n year = {2006},\\n pages = {287-297},\\n volume = {362},\\n id = {f66fd98b-e46d-356c-adc5-dbb7bb45a1c1},\\n created = {2023-01-10T01:45:55.259Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:55.259Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Gell, D.A. and Westman, B.J. and Gorman, D. and Liew, C. and Welch, J.J. and Weiss, M.J. and Mackay, J.P.},\\n doi = {10.1016/j.jmb.2006.07.010},\\n journal = {Journal of Molecular Biology},\\n number = {2}\\n}\",\"author_short\":[\"Gell,  D.\",\"Westman,  B.\",\"Gorman,  D.\",\"Liew,  C.\",\"Welch,  J.\",\"Weiss,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"GATA-1: One protein, many partners\",\"type\":\"article\",\"year\":\"2006\",\"pages\":\"6-11\",\"volume\":\"38\",\"id\":\"91c99222-cb02-3632-8751-7f2b89a9578c\",\"created\":\"2023-01-10T01:45:56.163Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:56.163Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Lowry, J.A. and MacKay, J.P.\",\"doi\":\"10.1016/j.biocel.2005.06.017\",\"journal\":\"International Journal of Biochemistry and Cell Biology\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {GATA-1: One protein, many partners},\\n type = {article},\\n year = {2006},\\n pages = {6-11},\\n volume = {38},\\n id = {91c99222-cb02-3632-8751-7f2b89a9578c},\\n created = {2023-01-10T01:45:56.163Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:56.163Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Lowry, J.A. and MacKay, J.P.},\\n doi = {10.1016/j.biocel.2005.06.017},\\n journal = {International Journal of Biochemistry and Cell Biology},\\n number = {1}\\n}\",\"author_short\":[\"Lowry,  J.\",\"MacKay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"lowry-mackay-gata1oneproteinmanypartners-2006\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Grb7-SH2 domain dimerisation is affected by a single point mutation\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"454-460\",\"volume\":\"34\",\"id\":\"273a7a2d-ac31-3984-b4ee-f7adf13fb08c\",\"created\":\"2023-01-10T01:45:57.071Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:57.071Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 μM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation. © EBSA 2005.\",\"bibtype\":\"article\",\"author\":\"Porter, C.J. and Wilce, M.C.J. and Mackay, J.P. and Leedman, P. and Wilce, J.A.\",\"doi\":\"10.1007/s00249-005-0480-1\",\"journal\":\"European Biophysics Journal\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Grb7-SH2 domain dimerisation is affected by a single point mutation},\\n type = {article},\\n year = {2005},\\n pages = {454-460},\\n volume = {34},\\n id = {273a7a2d-ac31-3984-b4ee-f7adf13fb08c},\\n created = {2023-01-10T01:45:57.071Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:57.071Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 μM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation. © EBSA 2005.},\\n bibtype = {article},\\n author = {Porter, C.J. and Wilce, M.C.J. and Mackay, J.P. and Leedman, P. and Wilce, J.A.},\\n doi = {10.1007/s00249-005-0480-1},\\n journal = {European Biophysics Journal},\\n number = {5}\\n}\",\"author_short\":[\"Porter,  C.\",\"Wilce,  M.\",\"Mackay,  J.\",\"Leedman,  P.\",\"Wilce,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structure of oxidized α-haemoglobin bound to AHSP reveals a protective mechanism for haem\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"697-701\",\"volume\":\"435\",\"id\":\"a2006ec4-0c0d-3521-8be7-b22dddf9fea3\",\"created\":\"2023-01-10T01:45:57.993Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:57.993Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The synthesis of haemoglobin A (HbA) is exquisitely coordinated during erythrocyte development to prevent damaging effects from individual α- and β-subunits. The α-haemoglobin-stabilizing protein (AHSP) binds α-haemoglobin (αHb), inhibits the ability of αHb to generate reactive oxygen species and prevents its precipitation on exposure to oxidant stress. The structure of AHSP bound to ferrous αHb is thought to represent a transitional complex through which αHb is converted to a non-reactive, hexacoordinate ferric form. Here we report the crystal structure of this ferric αHb-AHSP complex at 2.4 A resolution. Our findings reveal a striking bis-histidyl configuration in which both the proximal and the distal histidines coordinate the haem iron atom. To attain this unusual conformation, segments of αHb undergo drastic structural rearrangements, including the repositioning of several α-helices. Moreover, conversion to the ferric bishistidine configuration strongly and specifically inhibits redox chemistry catalysis and haem loss from αHb. The observed structural changes, which impair the chemical reactivity of haem iron, explain how AHSP stabilizes αHb and prevents its damaging effects in cells.\",\"bibtype\":\"article\",\"author\":\"Feng, L. and Zhou, S. and Gu, L. and Gell, D.A. and Mackay, J.P. and Weiss, M.J. and Gow, A.J. and Shi, Y.\",\"doi\":\"10.1038/nature03609\",\"journal\":\"Nature\",\"number\":\"7042\",\"bibtex\":\"@article{\\n title = {Structure of oxidized α-haemoglobin bound to AHSP reveals a protective mechanism for haem},\\n type = {article},\\n year = {2005},\\n pages = {697-701},\\n volume = {435},\\n id = {a2006ec4-0c0d-3521-8be7-b22dddf9fea3},\\n created = {2023-01-10T01:45:57.993Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:57.993Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The synthesis of haemoglobin A (HbA) is exquisitely coordinated during erythrocyte development to prevent damaging effects from individual α- and β-subunits. The α-haemoglobin-stabilizing protein (AHSP) binds α-haemoglobin (αHb), inhibits the ability of αHb to generate reactive oxygen species and prevents its precipitation on exposure to oxidant stress. The structure of AHSP bound to ferrous αHb is thought to represent a transitional complex through which αHb is converted to a non-reactive, hexacoordinate ferric form. Here we report the crystal structure of this ferric αHb-AHSP complex at 2.4 A resolution. Our findings reveal a striking bis-histidyl configuration in which both the proximal and the distal histidines coordinate the haem iron atom. To attain this unusual conformation, segments of αHb undergo drastic structural rearrangements, including the repositioning of several α-helices. Moreover, conversion to the ferric bishistidine configuration strongly and specifically inhibits redox chemistry catalysis and haem loss from αHb. The observed structural changes, which impair the chemical reactivity of haem iron, explain how AHSP stabilizes αHb and prevents its damaging effects in cells.},\\n bibtype = {article},\\n author = {Feng, L. and Zhou, S. and Gu, L. and Gell, D.A. and Mackay, J.P. and Weiss, M.J. and Gow, A.J. and Shi, Y.},\\n doi = {10.1038/nature03609},\\n journal = {Nature},\\n number = {7042}\\n}\",\"author_short\":[\"Feng,  L.\",\"Zhou,  S.\",\"Gu,  L.\",\"Gell,  D.\",\"Mackay,  J.\",\"Weiss,  M.\",\"Gow,  A.\",\"Shi,  Y.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"feng-zhou-gu-gell-mackay-weiss-gow-shi-structureofoxidizedhaemoglobinboundtoahsprevealsaprotectivemechanismforhaem-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular Lhx3:ldb1 complex [4]\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"198\",\"volume\":\"33\",\"id\":\"f9aa56cf-83b9-3151-a514-a77d7bddc008\",\"created\":\"2023-01-10T01:45:58.923Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:58.923Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Lee, C. and Nancarrow, A.L. and Bach, I. and Mackay, J.P. and Matthews, J.M.\",\"doi\":\"10.1007/s10858-005-3209-7\",\"journal\":\"Journal of Biomolecular NMR\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular Lhx3:ldb1 complex [4]},\\n type = {article},\\n year = {2005},\\n pages = {198},\\n volume = {33},\\n id = {f9aa56cf-83b9-3151-a514-a77d7bddc008},\\n created = {2023-01-10T01:45:58.923Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:58.923Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Lee, C. and Nancarrow, A.L. and Bach, I. and Mackay, J.P. and Matthews, J.M.},\\n doi = {10.1007/s10858-005-3209-7},\\n journal = {Journal of Biomolecular NMR},\\n number = {3}\\n}\",\"author_short\":[\"Lee,  C.\",\"Nancarrow,  A.\",\"Bach,  I.\",\"Mackay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"lee-nancarrow-bach-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlhx3ldb1complex4-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/friend of GATA interaction\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"583-588\",\"volume\":\"102\",\"id\":\"6ca8effa-53df-38c4-a21a-ded042ec2309\",\"created\":\"2023-01-10T01:45:59.921Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:45:59.921Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.\",\"bibtype\":\"article\",\"author\":\"Liew, C.K. and Simpson, R.J.Y. and Kwan, A.H.Y. and Crofts, L.A. and Loughlin, F.E. and Matthews, J.M. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1073/pnas.0407511102\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/friend of GATA interaction},\\n type = {article},\\n year = {2005},\\n pages = {583-588},\\n volume = {102},\\n id = {6ca8effa-53df-38c4-a21a-ded042ec2309},\\n created = {2023-01-10T01:45:59.921Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:45:59.921Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.},\\n bibtype = {article},\\n author = {Liew, C.K. and Simpson, R.J.Y. and Kwan, A.H.Y. and Crofts, L.A. and Loughlin, F.E. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1073/pnas.0407511102},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {3}\\n}\",\"author_short\":[\"Liew,  C.\",\"Simpson,  R.\",\"Kwan,  A.\",\"Crofts,  L.\",\"Loughlin,  F.\",\"Matthews,  J.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Assessment of the robustness of a serendipitous zinc binding fold: Mutagenesis and protein grafting\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"257-266\",\"volume\":\"13\",\"id\":\"9aeeb5d0-0dee-3dab-93d2-266c720ce66c\",\"created\":\"2023-01-10T01:46:00.845Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:00.845Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Zinc binding motifs have received much attention in the area of protein design. Here, we have tested the suitability of a recently discovered nonnative zinc binding structure as a protein design scaffold. A series of multiple alanine mutants was created to investigate the minimal requirements for folding, and solution structures of these mutants showed that the original fold was maintained, despite changes in 50% of the sequence. We next attempted to transplant binding faces from chosen bimolecular interactions onto one of these mutants, and many of the resulting \\\"chimeras\\\" were shown to adopt a native-like fold. These results both highlight the robust nature of small zinc binding domains and underscore the complexity of designing functional proteins, even using such small, highly ordered scaffolds as templates.\",\"bibtype\":\"article\",\"author\":\"Sharpe, B.K. and Liew, C.K. and Kwan, A.H. and Wilce, J.A. and Crossley, M. and Matthews, J.M. and Mackay, J.P.\",\"doi\":\"10.1016/j.str.2004.12.007\",\"journal\":\"Structure\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Assessment of the robustness of a serendipitous zinc binding fold: Mutagenesis and protein grafting},\\n type = {article},\\n year = {2005},\\n pages = {257-266},\\n volume = {13},\\n id = {9aeeb5d0-0dee-3dab-93d2-266c720ce66c},\\n created = {2023-01-10T01:46:00.845Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:00.845Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Zinc binding motifs have received much attention in the area of protein design. Here, we have tested the suitability of a recently discovered nonnative zinc binding structure as a protein design scaffold. A series of multiple alanine mutants was created to investigate the minimal requirements for folding, and solution structures of these mutants showed that the original fold was maintained, despite changes in 50% of the sequence. We next attempted to transplant binding faces from chosen bimolecular interactions onto one of these mutants, and many of the resulting \\\"chimeras\\\" were shown to adopt a native-like fold. These results both highlight the robust nature of small zinc binding domains and underscore the complexity of designing functional proteins, even using such small, highly ordered scaffolds as templates.},\\n bibtype = {article},\\n author = {Sharpe, B.K. and Liew, C.K. and Kwan, A.H. and Wilce, J.A. and Crossley, M. and Matthews, J.M. and Mackay, J.P.},\\n doi = {10.1016/j.str.2004.12.007},\\n journal = {Structure},\\n number = {2}\\n}\",\"author_short\":[\"Sharpe,  B.\",\"Liew,  C.\",\"Kwan,  A.\",\"Wilce,  J.\",\"Crossley,  M.\",\"Matthews,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"sharpe-liew-kwan-wilce-crossley-matthews-mackay-assessmentoftherobustnessofaserendipitouszincbindingfoldmutagenesisandproteingrafting-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution structure of a recombinant type I sculpin antifreeze protein\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"1980-1988\",\"volume\":\"44\",\"id\":\"02c3451d-5f79-3362-9ed9-2542509ffb84\",\"created\":\"2023-01-10T01:46:01.762Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:01.762Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"We have determined the solution structure of rSS3, a recombinant form of the type I shorthorn sculpin antifreeze protein (AFP), at 278 and 268 K. This AFP contains an unusual sequence of N-terminal residues, together with two of the 11-residue repeats that are characteristic of the type I winter flounder AFP. The solution conformation of the N-terminal region of the sculpin AFP has been assumed to be the critical factor that results in recognition of different ice planes by the sculpin and flounder AFPs. At 278 K, the two repeats units (residues 11-20 and 21-32) in rSS3 form a continuous α-helix, with the residues 30-33 in the second repeat somewhat less well defined. Within the N-terminal region, residues 2-6 are well defined and helical and linked to the main helix by a more flexible region comprising residues A7-T11. At 268 K the AFP is overall more helical but retains the apparent hinge region. The helical conformation of the two repeats units is almost identical to the corresponding repeats in the type I winter flounder AFP. We also show that while tetracetylated rSS3 has antifreeze activity comparable to the natural AFP, its overall structure is the same as that of the unacetylated peptide. These data provide some insight into the structural determinants of antifreeze activity and should assist in the development of models that explain the recognition of different ice interfaces by the sculpin and flounder type I AFPs.\",\"bibtype\":\"article\",\"author\":\"Kwan, A.H.-Y. and Fairley, K. and Anderberg, P.I. and Liew, C.W. and Harding, M.M. and Mackay, J.P.\",\"doi\":\"10.1021/bi047782j\",\"journal\":\"Biochemistry\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Solution structure of a recombinant type I sculpin antifreeze protein},\\n type = {article},\\n year = {2005},\\n pages = {1980-1988},\\n volume = {44},\\n id = {02c3451d-5f79-3362-9ed9-2542509ffb84},\\n created = {2023-01-10T01:46:01.762Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:01.762Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {We have determined the solution structure of rSS3, a recombinant form of the type I shorthorn sculpin antifreeze protein (AFP), at 278 and 268 K. This AFP contains an unusual sequence of N-terminal residues, together with two of the 11-residue repeats that are characteristic of the type I winter flounder AFP. The solution conformation of the N-terminal region of the sculpin AFP has been assumed to be the critical factor that results in recognition of different ice planes by the sculpin and flounder AFPs. At 278 K, the two repeats units (residues 11-20 and 21-32) in rSS3 form a continuous α-helix, with the residues 30-33 in the second repeat somewhat less well defined. Within the N-terminal region, residues 2-6 are well defined and helical and linked to the main helix by a more flexible region comprising residues A7-T11. At 268 K the AFP is overall more helical but retains the apparent hinge region. The helical conformation of the two repeats units is almost identical to the corresponding repeats in the type I winter flounder AFP. We also show that while tetracetylated rSS3 has antifreeze activity comparable to the natural AFP, its overall structure is the same as that of the unacetylated peptide. These data provide some insight into the structural determinants of antifreeze activity and should assist in the development of models that explain the recognition of different ice interfaces by the sculpin and flounder type I AFPs.},\\n bibtype = {article},\\n author = {Kwan, A.H.-Y. and Fairley, K. and Anderberg, P.I. and Liew, C.W. and Harding, M.M. and Mackay, J.P.},\\n doi = {10.1021/bi047782j},\\n journal = {Biochemistry},\\n number = {6}\\n}\",\"author_short\":[\"Kwan,  A.\",\"Fairley,  K.\",\"Anderberg,  P.\",\"Liew,  C.\",\"Harding,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kwan-fairley-anderberg-liew-harding-mackay-solutionstructureofarecombinanttypeisculpinantifreezeprotein-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Role of alpha hemoglobin-stabilizing protein in normal erythropoiesis and β-thalassemia\",\"type\":\"book\",\"year\":\"2005\",\"source\":\"Annals of the New York Academy of Sciences\",\"pages\":\"103-117\",\"volume\":\"1054\",\"id\":\"3eadb043-bae4-3de9-9a30-64811004299d\",\"created\":\"2023-01-10T01:46:02.725Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:02.725Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Hemoglobin (Hb) synthesis is coordinated by homeostatic mechanisms to limit the accumulation of free α or β subunits, which are cytotoxic. Alpha hemoglobin-stabilizing protein (AHSP) is an abundant erythroid protein that specifically binds free αHb, stabilizes its structure, and limits its ability to participate in chemical reactions that generate reactive oxygen species. Gene ablation studies in mice demonstrate that AHSP is required for normal erythropoiesis. AHSP-null erythrocytes are short-lived, contain Hb precipitates, and exhibit signs of oxidative damage. Loss of AHSP exacerbates β-thalassemia in mice, indicating that altered AHSP expression or function could modify thalassemia phenotypes in humans, a topic that is beginning to be explored in clinical studies. We used biochemical, spectroscopic, and crystallographic methods to examine how AHSP stabilizes αHb. AHSP binds the G and H helices of αHb on a surface that largely overlaps with the α1-β1 interface of HbA. This result explains previous findings that βHb can competitively displace AHSP from αHb to form HbA tetramer. Remarkably, binding of AHSP to oxygenated αHb induces dramatic conformational changes and converts the heme-bound iron to an oxidized hemichrome state in which all six coordinate positions are occupied. This structure limits the reactivity of heme iron, providing a mechanism by which AHSP stabilizes αHb. These findings suggest a biochemical pathway through which AHSP might participate in normal Hb synthesis and modulate the severity of thalassemias. Moreover, understanding how AHSP stabilizes αHb provides a theoretical basis for new strategies to inhibit the damaging effects of free αHb that accumulates in β-thalassemia. © 2005 New York Academy of Sciences.\",\"bibtype\":\"book\",\"author\":\"Weiss, M.J. and Zhou, S. and Feng, L. and Gell, D.A. and Mackay, J.P. and Shi, Y. and Gow, A.J.\",\"doi\":\"10.1196/annals.1345.013\",\"bibtex\":\"@book{\\n title = {Role of alpha hemoglobin-stabilizing protein in normal erythropoiesis and β-thalassemia},\\n type = {book},\\n year = {2005},\\n source = {Annals of the New York Academy of Sciences},\\n pages = {103-117},\\n volume = {1054},\\n id = {3eadb043-bae4-3de9-9a30-64811004299d},\\n created = {2023-01-10T01:46:02.725Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:02.725Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Hemoglobin (Hb) synthesis is coordinated by homeostatic mechanisms to limit the accumulation of free α or β subunits, which are cytotoxic. Alpha hemoglobin-stabilizing protein (AHSP) is an abundant erythroid protein that specifically binds free αHb, stabilizes its structure, and limits its ability to participate in chemical reactions that generate reactive oxygen species. Gene ablation studies in mice demonstrate that AHSP is required for normal erythropoiesis. AHSP-null erythrocytes are short-lived, contain Hb precipitates, and exhibit signs of oxidative damage. Loss of AHSP exacerbates β-thalassemia in mice, indicating that altered AHSP expression or function could modify thalassemia phenotypes in humans, a topic that is beginning to be explored in clinical studies. We used biochemical, spectroscopic, and crystallographic methods to examine how AHSP stabilizes αHb. AHSP binds the G and H helices of αHb on a surface that largely overlaps with the α1-β1 interface of HbA. This result explains previous findings that βHb can competitively displace AHSP from αHb to form HbA tetramer. Remarkably, binding of AHSP to oxygenated αHb induces dramatic conformational changes and converts the heme-bound iron to an oxidized hemichrome state in which all six coordinate positions are occupied. This structure limits the reactivity of heme iron, providing a mechanism by which AHSP stabilizes αHb. These findings suggest a biochemical pathway through which AHSP might participate in normal Hb synthesis and modulate the severity of thalassemias. Moreover, understanding how AHSP stabilizes αHb provides a theoretical basis for new strategies to inhibit the damaging effects of free αHb that accumulates in β-thalassemia. © 2005 New York Academy of Sciences.},\\n bibtype = {book},\\n author = {Weiss, M.J. and Zhou, S. and Feng, L. and Gell, D.A. and Mackay, J.P. and Shi, Y. and Gow, A.J.},\\n doi = {10.1196/annals.1345.013}\\n}\",\"author_short\":[\"Weiss,  M.\",\"Zhou,  S.\",\"Feng,  L.\",\"Gell,  D.\",\"Mackay,  J.\",\"Shi,  Y.\",\"Gow,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"weiss-zhou-feng-gell-mackay-shi-gow-roleofalphahemoglobinstabilizingproteininnormalerythropoiesisandthalassemia-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The hydrophobic domain 26 of human tropoelastin is unstructured in solution\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"154-162\",\"volume\":\"150\",\"id\":\"3e2d6e2e-f905-3566-81be-3cc4ef02fb5c\",\"created\":\"2023-01-10T01:46:03.625Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:03.625Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Elastin is the protein responsible for the elastic properties of vertebrate tissue. Very little is currently known about the structure of elastin or of its soluble precursor tropoelastin. We have used high-resolution solution NMR methods to probe the conformational preferences of a conserved hydrophobic region in tropoelastin, domain 26 (D26). Using a combination of homonuclear, 15N-separated and triple resonance experiments, we have obtained essentially full chemical shift assignments for D26 at 278 K. An analysis of secondary chemical shift changes, as well as NOE and 15N relaxation data, leads us to conclude that this domain is essentially unstructured in solution and does not interact with intact tropoelastin. D26 does not display exposed hydrophobic clusters, as expected for a fully unfolded protein and commensurate with an absence of flexible structural motifs, as identified by lack of binding of the fluorescent probe 4,4′-dianilino-1,1′- binaphthyl-5,5′-disulfonic acid. Sedimentation equilibrium data establish that this domain is strictly monomeric in solution. NMR spectra recorded at 278 and 308 K indicate that no significant structural changes occur for this domain over the temperature range 278-308 K, in contrast to the characteristic coacervation behavior that is observed for the full-length protein. © 2005 Elsevier Inc. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"MacKay, J.P. and Muiznieks, L.D. and Toonkool, P. and Weiss, A.S.\",\"doi\":\"10.1016/j.jsb.2005.02.005\",\"journal\":\"Journal of Structural Biology\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {The hydrophobic domain 26 of human tropoelastin is unstructured in solution},\\n type = {article},\\n year = {2005},\\n pages = {154-162},\\n volume = {150},\\n id = {3e2d6e2e-f905-3566-81be-3cc4ef02fb5c},\\n created = {2023-01-10T01:46:03.625Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:03.625Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Elastin is the protein responsible for the elastic properties of vertebrate tissue. Very little is currently known about the structure of elastin or of its soluble precursor tropoelastin. We have used high-resolution solution NMR methods to probe the conformational preferences of a conserved hydrophobic region in tropoelastin, domain 26 (D26). Using a combination of homonuclear, 15N-separated and triple resonance experiments, we have obtained essentially full chemical shift assignments for D26 at 278 K. An analysis of secondary chemical shift changes, as well as NOE and 15N relaxation data, leads us to conclude that this domain is essentially unstructured in solution and does not interact with intact tropoelastin. D26 does not display exposed hydrophobic clusters, as expected for a fully unfolded protein and commensurate with an absence of flexible structural motifs, as identified by lack of binding of the fluorescent probe 4,4′-dianilino-1,1′- binaphthyl-5,5′-disulfonic acid. Sedimentation equilibrium data establish that this domain is strictly monomeric in solution. NMR spectra recorded at 278 and 308 K indicate that no significant structural changes occur for this domain over the temperature range 278-308 K, in contrast to the characteristic coacervation behavior that is observed for the full-length protein. © 2005 Elsevier Inc. All rights reserved.},\\n bibtype = {article},\\n author = {MacKay, J.P. and Muiznieks, L.D. and Toonkool, P. and Weiss, A.S.},\\n doi = {10.1016/j.jsb.2005.02.005},\\n journal = {Journal of Structural Biology},\\n number = {2}\\n}\",\"author_short\":[\"MacKay,  J.\",\"Muiznieks,  L.\",\"Toonkool,  P.\",\"Weiss,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-muiznieks-toonkool-weiss-thehydrophobicdomain26ofhumantropoelastinisunstructuredinsolution-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A complex mechanism determines polarity of DNA replication fork arrest by the replication terminator complex of Bacillus subtilis\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"13105-13113\",\"volume\":\"280\",\"id\":\"826a04da-c81b-3dcb-bff3-7b7d2e7fede1\",\"created\":\"2023-01-10T01:46:04.549Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:04.549Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Two dimers of the replication terminator protein (RTP) of Bacillus subtilis bind to a chromosomal DNA terminator site to effect polar replication fork arrest. Cooperative binding of the dimers to overlapping half-sites within the terminator is essential for arrest. It was suggested previously that polarity of fork arrest is the result of the RTP dimer at the blocking (proximal) side within the complex binding very tightly and the permissive-side RTP dimer binding relatively weakly. In order to investigate this \\\"differential binding affinity\\\" model, we have constructed a series of mutant terminators that contain half-sites of widely different RTP binding affinities in various combinations. Although there appeared to be a correlation between binding affinity at the proximal half-site and fork arrest efficiency in vivo for some terminators, several deviated significantly from this correlation. Some terminators exhibited greatly reduced binding cooperativity (and therefore have reduced affinity at each half-site) but were highly efficient in fork arrest, whereas one terminator had normal affinity over the proximal half-site, yet had low fork arrest efficiency. The results show clearly that there is no direct correlation between the RTP binding affinity (either within the full complex or at the proximal half-site within the full complex) and the efficiency of replication fork arrest in vivo. Thus, the differential binding affinity over the proximal and distal half-sites cannot be solely responsible for functional polarity of fork arrest. Furthermore, efficient fork arrest relies on features in addition to the tight binding of RTP to terminator DNA. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc.\",\"bibtype\":\"article\",\"author\":\"Duggin, I.G. and Matthews, J.M. and Dixon, N.E. and Wake, R.G. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M414187200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"13\",\"bibtex\":\"@article{\\n title = {A complex mechanism determines polarity of DNA replication fork arrest by the replication terminator complex of Bacillus subtilis},\\n type = {article},\\n year = {2005},\\n pages = {13105-13113},\\n volume = {280},\\n id = {826a04da-c81b-3dcb-bff3-7b7d2e7fede1},\\n created = {2023-01-10T01:46:04.549Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:04.549Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Two dimers of the replication terminator protein (RTP) of Bacillus subtilis bind to a chromosomal DNA terminator site to effect polar replication fork arrest. Cooperative binding of the dimers to overlapping half-sites within the terminator is essential for arrest. It was suggested previously that polarity of fork arrest is the result of the RTP dimer at the blocking (proximal) side within the complex binding very tightly and the permissive-side RTP dimer binding relatively weakly. In order to investigate this \\\"differential binding affinity\\\" model, we have constructed a series of mutant terminators that contain half-sites of widely different RTP binding affinities in various combinations. Although there appeared to be a correlation between binding affinity at the proximal half-site and fork arrest efficiency in vivo for some terminators, several deviated significantly from this correlation. Some terminators exhibited greatly reduced binding cooperativity (and therefore have reduced affinity at each half-site) but were highly efficient in fork arrest, whereas one terminator had normal affinity over the proximal half-site, yet had low fork arrest efficiency. The results show clearly that there is no direct correlation between the RTP binding affinity (either within the full complex or at the proximal half-site within the full complex) and the efficiency of replication fork arrest in vivo. Thus, the differential binding affinity over the proximal and distal half-sites cannot be solely responsible for functional polarity of fork arrest. Furthermore, efficient fork arrest relies on features in addition to the tight binding of RTP to terminator DNA. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc.},\\n bibtype = {article},\\n author = {Duggin, I.G. and Matthews, J.M. and Dixon, N.E. and Wake, R.G. and Mackay, J.P.},\\n doi = {10.1074/jbc.M414187200},\\n journal = {Journal of Biological Chemistry},\\n number = {13}\\n}\",\"author_short\":[\"Duggin,  I.\",\"Matthews,  J.\",\"Dixon,  N.\",\"Wake,  R.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"duggin-matthews-dixon-wake-mackay-acomplexmechanismdeterminespolarityofdnareplicationforkarrestbythereplicationterminatorcomplexofbacillussubtilis-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"CSL: A notch above the rest\",\"type\":\"article\",\"year\":\"2005\",\"pages\":\"2472-2477\",\"volume\":\"37\",\"id\":\"951f9e11-35e5-393f-985a-6df0a62be9ff\",\"created\":\"2023-01-10T01:46:05.458Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:05.458Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"CSL (CBF1, Suppressor of Hairless, Lag-1) is a transcription factor that is responsible for activating the genes downstream of the Notch signalling pathway, a pathway that is essential for the development of the nervous system and the differentiation of the haematopoietic system among others. In the absence of Notch signalling, CSL represses transcription of Notch target genes, and following activation by Notch, CSL is converted into a transcriptional activator and activates transcription of the same genes. These two opposing functions of CSL are mediated through interactions with distinct protein complexes. The Notch signalling pathway and its crucial cofactor CSL can maintain cells in an undifferentiated state, and have therefore been associated with a growing list of cancers. In addition, CSL has been co-opted by Epstein-Barr virus to mediate viral and host gene transcription following infection. © 2005 Elsevier Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Pursglove, S.E. and Mackay, J.P.\",\"doi\":\"10.1016/j.biocel.2005.06.013\",\"journal\":\"International Journal of Biochemistry and Cell Biology\",\"number\":\"12\",\"bibtex\":\"@article{\\n title = {CSL: A notch above the rest},\\n type = {article},\\n year = {2005},\\n pages = {2472-2477},\\n volume = {37},\\n id = {951f9e11-35e5-393f-985a-6df0a62be9ff},\\n created = {2023-01-10T01:46:05.458Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:05.458Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {CSL (CBF1, Suppressor of Hairless, Lag-1) is a transcription factor that is responsible for activating the genes downstream of the Notch signalling pathway, a pathway that is essential for the development of the nervous system and the differentiation of the haematopoietic system among others. In the absence of Notch signalling, CSL represses transcription of Notch target genes, and following activation by Notch, CSL is converted into a transcriptional activator and activates transcription of the same genes. These two opposing functions of CSL are mediated through interactions with distinct protein complexes. The Notch signalling pathway and its crucial cofactor CSL can maintain cells in an undifferentiated state, and have therefore been associated with a growing list of cancers. In addition, CSL has been co-opted by Epstein-Barr virus to mediate viral and host gene transcription following infection. © 2005 Elsevier Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Pursglove, S.E. and Mackay, J.P.},\\n doi = {10.1016/j.biocel.2005.06.013},\\n journal = {International Journal of Biochemistry and Cell Biology},\\n number = {12}\\n}\",\"author_short\":[\"Pursglove,  S.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"pursglove-mackay-cslanotchabovetherest-2005\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Molecular mechanism of AHSP-mediated stabilization of α-hemoglobin\",\"type\":\"article\",\"year\":\"2004\",\"pages\":\"629-640\",\"volume\":\"119\",\"id\":\"ed2d19cb-f91e-3151-b6da-974c5bb50868\",\"created\":\"2023-01-10T01:46:06.370Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:06.370Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Hemoglobin A (HbA), the oxygen delivery system in humans, comprises two α and two β subunits. Free α-hemoglobin (αHb) is unstable, and its precipitation contributes to the pathophysiology of β thalassemia. In erythrocytes, the α-hemoglobin stabilizing protein (AHSP) binds αHb and inhibits its precipitation. The crystal structure of AHSP bound to Fe(II)-αHb reveals that AHSP specifically recognizes the G and H helices of αHb through a hydrophobic interface that largely recapitulates the α1-β1 interface of hemoglobin. The AHSP-αHb interactions are extensive but suboptimal, explaining why β-hemoglobin can competitively displace AHSP to form HbA. Remarkably, the Fe(II)-heme group in AHSP bound αHb is coordinated by the distal but not the proximal histidine. Importantly, binding to AHSP facilitates the conversion of oxy-αHb to a deoxygenated, oxidized [Fe(III)], nonreactive form in which all six coordinate positions are occupied. These observations reveal the molecular mechanisms by which AHSP stabilizes free αHb.\",\"bibtype\":\"article\",\"author\":\"Feng, L. and Gell, D.A. and Zhou, S. and Gu, L. and Kong, Y. and Li, J. and Hu, M. and Yan, N. and Lee, C. and Rich, A.M. and MacKay, J.P. and Shi, Y.\",\"doi\":\"10.1016/j.cell.2004.11.025\",\"journal\":\"Cell\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {Molecular mechanism of AHSP-mediated stabilization of α-hemoglobin},\\n type = {article},\\n year = {2004},\\n pages = {629-640},\\n volume = {119},\\n id = {ed2d19cb-f91e-3151-b6da-974c5bb50868},\\n created = {2023-01-10T01:46:06.370Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:06.370Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Hemoglobin A (HbA), the oxygen delivery system in humans, comprises two α and two β subunits. Free α-hemoglobin (αHb) is unstable, and its precipitation contributes to the pathophysiology of β thalassemia. In erythrocytes, the α-hemoglobin stabilizing protein (AHSP) binds αHb and inhibits its precipitation. The crystal structure of AHSP bound to Fe(II)-αHb reveals that AHSP specifically recognizes the G and H helices of αHb through a hydrophobic interface that largely recapitulates the α1-β1 interface of hemoglobin. The AHSP-αHb interactions are extensive but suboptimal, explaining why β-hemoglobin can competitively displace AHSP to form HbA. Remarkably, the Fe(II)-heme group in AHSP bound αHb is coordinated by the distal but not the proximal histidine. Importantly, binding to AHSP facilitates the conversion of oxy-αHb to a deoxygenated, oxidized [Fe(III)], nonreactive form in which all six coordinate positions are occupied. These observations reveal the molecular mechanisms by which AHSP stabilizes free αHb.},\\n bibtype = {article},\\n author = {Feng, L. and Gell, D.A. and Zhou, S. and Gu, L. and Kong, Y. and Li, J. and Hu, M. and Yan, N. and Lee, C. and Rich, A.M. and MacKay, J.P. and Shi, Y.},\\n doi = {10.1016/j.cell.2004.11.025},\\n journal = {Cell},\\n number = {5}\\n}\",\"author_short\":[\"Feng,  L.\",\"Gell,  D.\",\"Zhou,  S.\",\"Gu,  L.\",\"Kong,  Y.\",\"Li,  J.\",\"Hu,  M.\",\"Yan,  N.\",\"Lee,  C.\",\"Rich,  A.\",\"MacKay,  J.\",\"Shi,  Y.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"feng-gell-zhou-gu-kong-li-hu-yan-etal-molecularmechanismofahspmediatedstabilizationofhemoglobin-2004\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Classic zinc finger from friend of GATA mediates an interaction with the coiled-coil of transforming acidic coiled-coil 3\",\"type\":\"article\",\"year\":\"2004\",\"pages\":\"39789-39797\",\"volume\":\"279\",\"id\":\"8ae82118-628a-3d0f-85fb-40ef4ede1b72\",\"created\":\"2023-01-10T01:46:07.273Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:07.273Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Classic zinc finger domains (cZFs) consist of a β-hairpin followed by an α-helix. They are among the most abundant of all protein domains and are often found in tandem arrays in DNA-binding proteins, with each finger contributing an α-helix to effect sequence-specific DNA recognition. Lone cZFs, not found in tandem arrays, have been postulated to function in protein interactions. We have studied the transcriptional co-regulator Friend of GATA (FOG), which contains nine zinc fingers. We have discovered that the third cZF of FOG contacts a coiled-coil domain in the centrosomal protein transforming acidic coiled-coil 3 (TACC3). Although FOG-ZF3 exhibited low solubility, we have used a combination of mutational mapping and protein engineering to generate a derivative that was suitable for in vitro and structural analysis. We report that the α-helix of FOG-ZF3 recognizes a C-terminal portion of the TACC3 coiled-coil. Remarkably, the α-helical surface utilized by FOG-ZF3 is the same surface responsible for the well established sequence-specific DNA-binding properties of many other cZFs. Our data demonstrate the versatility of cZFs and have implications for the analysis of many as yet uncharacterized cZF proteins.\",\"bibtype\":\"article\",\"author\":\"Simpson, R.J.Y. and Lee, S.H.Y. and Bartle, N. and Sum, E.Y. and Visvader, J.E. and Matthews, J.M. and Mackay, J.P. and Crossley, M.\",\"doi\":\"10.1074/jbc.M404130200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"38\",\"bibtex\":\"@article{\\n title = {Classic zinc finger from friend of GATA mediates an interaction with the coiled-coil of transforming acidic coiled-coil 3},\\n type = {article},\\n year = {2004},\\n pages = {39789-39797},\\n volume = {279},\\n id = {8ae82118-628a-3d0f-85fb-40ef4ede1b72},\\n created = {2023-01-10T01:46:07.273Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:07.273Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Classic zinc finger domains (cZFs) consist of a β-hairpin followed by an α-helix. They are among the most abundant of all protein domains and are often found in tandem arrays in DNA-binding proteins, with each finger contributing an α-helix to effect sequence-specific DNA recognition. Lone cZFs, not found in tandem arrays, have been postulated to function in protein interactions. We have studied the transcriptional co-regulator Friend of GATA (FOG), which contains nine zinc fingers. We have discovered that the third cZF of FOG contacts a coiled-coil domain in the centrosomal protein transforming acidic coiled-coil 3 (TACC3). Although FOG-ZF3 exhibited low solubility, we have used a combination of mutational mapping and protein engineering to generate a derivative that was suitable for in vitro and structural analysis. We report that the α-helix of FOG-ZF3 recognizes a C-terminal portion of the TACC3 coiled-coil. Remarkably, the α-helical surface utilized by FOG-ZF3 is the same surface responsible for the well established sequence-specific DNA-binding properties of many other cZFs. Our data demonstrate the versatility of cZFs and have implications for the analysis of many as yet uncharacterized cZF proteins.},\\n bibtype = {article},\\n author = {Simpson, R.J.Y. and Lee, S.H.Y. and Bartle, N. and Sum, E.Y. and Visvader, J.E. and Matthews, J.M. and Mackay, J.P. and Crossley, M.},\\n doi = {10.1074/jbc.M404130200},\\n journal = {Journal of Biological Chemistry},\\n number = {38}\\n}\",\"author_short\":[\"Simpson,  R.\",\"Lee,  S.\",\"Bartle,  N.\",\"Sum,  E.\",\"Visvader,  J.\",\"Matthews,  J.\",\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"simpson-lee-bartle-sum-visvader-matthews-mackay-crossley-classiczincfingerfromfriendofgatamediatesaninteractionwiththecoiledcoiloftransformingacidiccoiledcoil3-2004\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Presence of transient helical segments in the galanin-like peptide evident from <sup>1</sup>H NMR, circular dichroism, and prediction studies\",\"type\":\"article\",\"year\":\"2004\",\"pages\":\"261-271\",\"volume\":\"146\",\"id\":\"dcdf6b5e-2e76-3021-abce-66a56e5b7f57\",\"created\":\"2023-01-10T01:46:08.193Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:08.193Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods. © 2004 Elsevier Inc. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Dastmalchi, S. and Church, W.B. and Morris, M.B. and Iismaa, T.P. and Mackay, J.P.\",\"doi\":\"10.1016/j.jsb.2004.01.004\",\"journal\":\"Journal of Structural Biology\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Presence of transient helical segments in the galanin-like peptide evident from <sup>1</sup>H NMR, circular dichroism, and prediction studies},\\n type = {article},\\n year = {2004},\\n pages = {261-271},\\n volume = {146},\\n id = {dcdf6b5e-2e76-3021-abce-66a56e5b7f57},\\n created = {2023-01-10T01:46:08.193Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:08.193Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods. © 2004 Elsevier Inc. All rights reserved.},\\n bibtype = {article},\\n author = {Dastmalchi, S. and Church, W.B. and Morris, M.B. and Iismaa, T.P. and Mackay, J.P.},\\n doi = {10.1016/j.jsb.2004.01.004},\\n journal = {Journal of Structural Biology},\\n number = {3}\\n}\",\"author_short\":[\"Dastmalchi,  S.\",\"Church,  W.\",\"Morris,  M.\",\"Iismaa,  T.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfromsup1suphnmrcirculardichroismandpredictionstudies-2004\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural and functional analysis of the Josephin domain of the polyglutamine protein ataxin-3\",\"type\":\"article\",\"year\":\"2004\",\"pages\":\"387-394\",\"volume\":\"322\",\"id\":\"74e20e29-48a8-34da-ac85-4beac421f531\",\"created\":\"2023-01-10T01:46:09.118Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:09.118Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Ataxin-3 belongs to the family of polyglutamine proteins, which are associated with nine different neurodegenerative disorders. Relatively little is known about the structural and functional properties of ataxin-3, and only recently have these aspects of the protein begun to be explored. We have performed a preliminary investigation into the conserved N-terminal domain of ataxin-3, termed Josephin. We show that Josephin is a monomeric domain which folds into a globular conformation and possesses ubiquitin protease activity. In addition, we demonstrate that the presence of the polyglutamine region of the protein does not alter the structure of the protein. However, its presence destabilizes the Josephin domain. The implications of these data in the pathogenesis of polyglutamine repeat proteins are discussed. © 2004 Elsevier Inc. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Chow, M.K.M. and MacKay, J.P. and Whisstock, J.C. and Scanlon, M.J. and Bottomley, S.P.\",\"doi\":\"10.1016/j.bbrc.2004.07.131\",\"journal\":\"Biochemical and Biophysical Research Communications\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Structural and functional analysis of the Josephin domain of the polyglutamine protein ataxin-3},\\n type = {article},\\n year = {2004},\\n pages = {387-394},\\n volume = {322},\\n id = {74e20e29-48a8-34da-ac85-4beac421f531},\\n created = {2023-01-10T01:46:09.118Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:09.118Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Ataxin-3 belongs to the family of polyglutamine proteins, which are associated with nine different neurodegenerative disorders. Relatively little is known about the structural and functional properties of ataxin-3, and only recently have these aspects of the protein begun to be explored. We have performed a preliminary investigation into the conserved N-terminal domain of ataxin-3, termed Josephin. We show that Josephin is a monomeric domain which folds into a globular conformation and possesses ubiquitin protease activity. In addition, we demonstrate that the presence of the polyglutamine region of the protein does not alter the structure of the protein. However, its presence destabilizes the Josephin domain. The implications of these data in the pathogenesis of polyglutamine repeat proteins are discussed. © 2004 Elsevier Inc. All rights reserved.},\\n bibtype = {article},\\n author = {Chow, M.K.M. and MacKay, J.P. and Whisstock, J.C. and Scanlon, M.J. and Bottomley, S.P.},\\n doi = {10.1016/j.bbrc.2004.07.131},\\n journal = {Biochemical and Biophysical Research Communications},\\n number = {2}\\n}\",\"author_short\":[\"Chow,  M.\",\"MacKay,  J.\",\"Whisstock,  J.\",\"Scanlon,  M.\",\"Bottomley,  S.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"chow-mackay-whisstock-scanlon-bottomley-structuralandfunctionalanalysisofthejosephindomainofthepolyglutamineproteinataxin3-2004\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Crystal and Solution Structures of a Superantigen from Yersinia pseudotuberculosis Reveal a Yelly-Roll Fold\",\"type\":\"article\",\"year\":\"2004\",\"pages\":\"145-156\",\"volume\":\"12\",\"id\":\"cff79b19-ac8f-3b35-9aa3-73d032e0181d\",\"created\":\"2023-01-10T01:46:10.033Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:10.033Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Superantigens are a class of microbial proteins with the ability to excessively activate T cells by binding to the T cell receptor. The staphylococcal and streptococcal superantigens are closely related in structure and possess an N-terminal domain that resembles an OB fold and a C-terminal domain similar to a β-grasp fold. Yersinia pseudotuberculosis produces superantigens, YPMa, YPMb, and YPMc, which have no significant amino acid similarity to other proteins. We have determined the crystal and solution structures of YPMa, which show that the protein has a jelly-roll fold. The closest structural neighbors to YPMa are viral capsid proteins and members of the tumor necrosis factor superfamily. In the crystal structure, YPMa packs as a trimer, another feature shared with viral capsid proteins and TNF superfamily proteins. However, in solution YPMa behaves as a monomer, and any functional relevance of the trimer observed in the crystals is yet to be established.\",\"bibtype\":\"article\",\"author\":\"Donadini, R. and Liew, C.W. and Kwan, A.H.Y. and Mackay, J.P. and Fields, B.A.\",\"doi\":\"10.1016/j.str.2003.12.002\",\"journal\":\"Structure\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Crystal and Solution Structures of a Superantigen from Yersinia pseudotuberculosis Reveal a Yelly-Roll Fold},\\n type = {article},\\n year = {2004},\\n pages = {145-156},\\n volume = {12},\\n id = {cff79b19-ac8f-3b35-9aa3-73d032e0181d},\\n created = {2023-01-10T01:46:10.033Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:10.033Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Superantigens are a class of microbial proteins with the ability to excessively activate T cells by binding to the T cell receptor. The staphylococcal and streptococcal superantigens are closely related in structure and possess an N-terminal domain that resembles an OB fold and a C-terminal domain similar to a β-grasp fold. Yersinia pseudotuberculosis produces superantigens, YPMa, YPMb, and YPMc, which have no significant amino acid similarity to other proteins. We have determined the crystal and solution structures of YPMa, which show that the protein has a jelly-roll fold. The closest structural neighbors to YPMa are viral capsid proteins and members of the tumor necrosis factor superfamily. In the crystal structure, YPMa packs as a trimer, another feature shared with viral capsid proteins and TNF superfamily proteins. However, in solution YPMa behaves as a monomer, and any functional relevance of the trimer observed in the crystals is yet to be established.},\\n bibtype = {article},\\n author = {Donadini, R. and Liew, C.W. and Kwan, A.H.Y. and Mackay, J.P. and Fields, B.A.},\\n doi = {10.1016/j.str.2003.12.002},\\n journal = {Structure},\\n number = {1}\\n}\",\"author_short\":[\"Donadini,  R.\",\"Liew,  C.\",\"Kwan,  A.\",\"Mackay,  J.\",\"Fields,  B.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"donadini-liew-kwan-mackay-fields-crystalandsolutionstructuresofasuperantigenfromyersiniapseudotuberculosisrevealayellyrollfold-2004\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Loss of α-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates β-thalassemia\",\"type\":\"article\",\"year\":\"2004\",\"pages\":\"1457-1466\",\"volume\":\"114\",\"id\":\"b699fcb7-add4-3ff6-a946-b7bf6448eaeb\",\"created\":\"2023-01-10T01:46:10.944Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:10.944Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Hemoglobin (Hb) A production during red blood cell development is coordinated to minimize the deleterious effects of free α- and β-Hb subunits, which are unstable and cytotoxic. The α-Hb-stabilizing protein (AHSP) is an erythroid protein that specifically binds α-Hb and prevents its precipitation in vitro, which suggests that it may function to limit free α-Hb toxicities in vivo. We investigated this possibility through gene ablation and biochemical studies. AHSP-/- erythrocytes contained hemoglobin precipitates and were short-lived. In hematopoietic tissues, erythroid precursors were elevated in number but exhibited increased apoptosis. Consistent with unstable α-Hb, AHSP-/- erythrocytes contained increased ROS and evidence of oxidative damage. Moreover, purified recombinant AHSP inhibited ROS production by α-Hb in solution. Finally, loss of AHSP worsened the phenotype of β-thalassemia, a common inherited anemia characterized by excess free α-Hb. Together, the data support a model in which AHSP binds α-Hb transiently to stabilize its conformation and render it biochemically inert prior to Hb A assembly. This function is essential for normal erythropoiesis and, to a greater extent, in β-thalassemia. Our findings raise the possibility that altered AHSP expression levels could modulate the severity of β-thalassemia in humans.\",\"bibtype\":\"article\",\"author\":\"Kong, Y. and Zhou, S. and Kihm, A.J. and Katein, A.M. and Yu, X. and Gell, D.A. and Mackay, J.P. and Adachi, K. and Foster-Brown, L. and Louden, C.S. and Gow, A.J. and Weiss, M.J.\",\"doi\":\"10.1172/JCI21982\",\"journal\":\"Journal of Clinical Investigation\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {Loss of α-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates β-thalassemia},\\n type = {article},\\n year = {2004},\\n pages = {1457-1466},\\n volume = {114},\\n id = {b699fcb7-add4-3ff6-a946-b7bf6448eaeb},\\n created = {2023-01-10T01:46:10.944Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:10.944Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Hemoglobin (Hb) A production during red blood cell development is coordinated to minimize the deleterious effects of free α- and β-Hb subunits, which are unstable and cytotoxic. The α-Hb-stabilizing protein (AHSP) is an erythroid protein that specifically binds α-Hb and prevents its precipitation in vitro, which suggests that it may function to limit free α-Hb toxicities in vivo. We investigated this possibility through gene ablation and biochemical studies. AHSP-/- erythrocytes contained hemoglobin precipitates and were short-lived. In hematopoietic tissues, erythroid precursors were elevated in number but exhibited increased apoptosis. Consistent with unstable α-Hb, AHSP-/- erythrocytes contained increased ROS and evidence of oxidative damage. Moreover, purified recombinant AHSP inhibited ROS production by α-Hb in solution. Finally, loss of AHSP worsened the phenotype of β-thalassemia, a common inherited anemia characterized by excess free α-Hb. Together, the data support a model in which AHSP binds α-Hb transiently to stabilize its conformation and render it biochemically inert prior to Hb A assembly. This function is essential for normal erythropoiesis and, to a greater extent, in β-thalassemia. Our findings raise the possibility that altered AHSP expression levels could modulate the severity of β-thalassemia in humans.},\\n bibtype = {article},\\n author = {Kong, Y. and Zhou, S. and Kihm, A.J. and Katein, A.M. and Yu, X. and Gell, D.A. and Mackay, J.P. and Adachi, K. and Foster-Brown, L. and Louden, C.S. and Gow, A.J. and Weiss, M.J.},\\n doi = {10.1172/JCI21982},\\n journal = {Journal of Clinical Investigation},\\n number = {10}\\n}\",\"author_short\":[\"Kong,  Y.\",\"Zhou,  S.\",\"Kihm,  A.\",\"Katein,  A.\",\"Yu,  X.\",\"Gell,  D.\",\"Mackay,  J.\",\"Adachi,  K.\",\"Foster-Brown,  L.\",\"Louden,  C.\",\"Gow,  A.\",\"Weiss,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kong-zhou-kihm-katein-yu-gell-mackay-adachi-etal-lossofhemoglobinstabilizingproteinimpairserythropoiesisandexacerbatesthalassemia-2004\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Structural studies on a protein-binding zinc-finger domain of eos reveal both similarities and differences to classical zinc fingers\",\"type\":\"article\",\"year\":\"2004\",\"pages\":\"13318-13327\",\"volume\":\"43\",\"id\":\"08b79ba5-e49f-31ea-9efb-6f22acd29fae\",\"created\":\"2023-01-10T01:46:11.867Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:11.867Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The oligomerization domain that is present at the C terminus of Ikaros-family proteins and the protein Trps-1 is important for the proper regulation of developmental processes such as hematopoiesis. Remarkably, this domain is predicted to contain two classical zinc fingers (ZnFs), domains normally associated with the recognition of nucleic acids. The preference for protein binding by these predicted ZnFs is not well-understood. We have used a range of methods to gain insight into the structure of this domain. Circular dichroism, UV - vis, and NMR experiments carried out on the C-terminal domain of Eos (EosC) revealed that the two putative ZnFs (C1 and C2) are separable, i.e., capable of folding independently in the presence of ZnII. We next determined the structure of EosC2 using NMR spectroscopy, revealing that, although the overall fold of EosC2 is similar to other classical ZnFs, a number of differences exist. For example, the conformation of the C terminus of EosC2 appears to be flexible and may result in a major rearrangement of the zinc ligands. Finally, alanine-scanning mutagenesis was used to identify the residues that are involved in the homo- and hetero-oligomerization of Eos, and these results are discussed in the context of the structure of EosC. These studies provide the first structural insights into how EosC mediates protein - protein interactions and contributes to our understanding of why it does not exhibit high-affinity DNA binding.\",\"bibtype\":\"article\",\"author\":\"Westman, B.J. and Perdomo, J. and Matthews, J.M. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1021/bi049506a\",\"journal\":\"Biochemistry\",\"number\":\"42\",\"bibtex\":\"@article{\\n title = {Structural studies on a protein-binding zinc-finger domain of eos reveal both similarities and differences to classical zinc fingers},\\n type = {article},\\n year = {2004},\\n pages = {13318-13327},\\n volume = {43},\\n id = {08b79ba5-e49f-31ea-9efb-6f22acd29fae},\\n created = {2023-01-10T01:46:11.867Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:11.867Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The oligomerization domain that is present at the C terminus of Ikaros-family proteins and the protein Trps-1 is important for the proper regulation of developmental processes such as hematopoiesis. Remarkably, this domain is predicted to contain two classical zinc fingers (ZnFs), domains normally associated with the recognition of nucleic acids. The preference for protein binding by these predicted ZnFs is not well-understood. We have used a range of methods to gain insight into the structure of this domain. Circular dichroism, UV - vis, and NMR experiments carried out on the C-terminal domain of Eos (EosC) revealed that the two putative ZnFs (C1 and C2) are separable, i.e., capable of folding independently in the presence of ZnII. We next determined the structure of EosC2 using NMR spectroscopy, revealing that, although the overall fold of EosC2 is similar to other classical ZnFs, a number of differences exist. For example, the conformation of the C terminus of EosC2 appears to be flexible and may result in a major rearrangement of the zinc ligands. Finally, alanine-scanning mutagenesis was used to identify the residues that are involved in the homo- and hetero-oligomerization of Eos, and these results are discussed in the context of the structure of EosC. These studies provide the first structural insights into how EosC mediates protein - protein interactions and contributes to our understanding of why it does not exhibit high-affinity DNA binding.},\\n bibtype = {article},\\n author = {Westman, B.J. and Perdomo, J. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1021/bi049506a},\\n journal = {Biochemistry},\\n number = {42}\\n}\",\"author_short\":[\"Westman,  B.\",\"Perdomo,  J.\",\"Matthews,  J.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"westman-perdomo-matthews-crossley-mackay-structuralstudiesonaproteinbindingzincfingerdomainofeosrevealbothsimilaritiesanddifferencestoclassicalzincfingers-2004\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"TC-1 Is a Novel Tumorigenic and Natively Disordered Protein Associated with Thyroid Cancer\",\"type\":\"article\",\"year\":\"2004\",\"pages\":\"2766-2773\",\"volume\":\"64\",\"id\":\"17d8f871-48a9-351a-8a2f-85f939c4f286\",\"created\":\"2023-01-10T01:46:12.786Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:12.786Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"A novel gene, thyroid cancer 1 (TC-1), was found recently to be overexpressed in thyroid cancer. TC-1 shows no homology to any of the known thyroid cancer-associated genes. We have produced stable transformants of normal thyroid cells that express the TC-1 gene, and these cells show increased proliferation rates and anchorage-independent growth in soft agar. Apoptosis rates also are decreased in the transformed cells. We also have expressed recombinant TC-1 protein and have undertaken a structural and functional characterization of the protein. The protein is monomeric and predominantly unstructured under conditions of physiologic salt and pH. This places it in the category of natively disordered proteins, a rapidly expanding group of proteins, many members of which play critical roles in cell regulation processes. We show that the protein can be phosphorylated by cyclic AMP-dependent protein kinase and protein kinase C, and the activity of both of these kinases is up-regulated when cells are stably transfected with TC-1. These results suggest that overexpression of TC-1 may be important in thyroid carcinogenesis.\",\"bibtype\":\"article\",\"author\":\"Sunde, M. and McGrath, K.C.Y. and Young, L. and Matthews, J.M. and Chua, E.L. and Mackay, J.P. and Death, A.K.\",\"doi\":\"10.1158/0008-5472.CAN-03-2093\",\"journal\":\"Cancer Research\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {TC-1 Is a Novel Tumorigenic and Natively Disordered Protein Associated with Thyroid Cancer},\\n type = {article},\\n year = {2004},\\n pages = {2766-2773},\\n volume = {64},\\n id = {17d8f871-48a9-351a-8a2f-85f939c4f286},\\n created = {2023-01-10T01:46:12.786Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:12.786Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {A novel gene, thyroid cancer 1 (TC-1), was found recently to be overexpressed in thyroid cancer. TC-1 shows no homology to any of the known thyroid cancer-associated genes. We have produced stable transformants of normal thyroid cells that express the TC-1 gene, and these cells show increased proliferation rates and anchorage-independent growth in soft agar. Apoptosis rates also are decreased in the transformed cells. We also have expressed recombinant TC-1 protein and have undertaken a structural and functional characterization of the protein. The protein is monomeric and predominantly unstructured under conditions of physiologic salt and pH. This places it in the category of natively disordered proteins, a rapidly expanding group of proteins, many members of which play critical roles in cell regulation processes. We show that the protein can be phosphorylated by cyclic AMP-dependent protein kinase and protein kinase C, and the activity of both of these kinases is up-regulated when cells are stably transfected with TC-1. These results suggest that overexpression of TC-1 may be important in thyroid carcinogenesis.},\\n bibtype = {article},\\n author = {Sunde, M. and McGrath, K.C.Y. and Young, L. and Matthews, J.M. and Chua, E.L. and Mackay, J.P. and Death, A.K.},\\n doi = {10.1158/0008-5472.CAN-03-2093},\\n journal = {Cancer Research},\\n number = {8}\\n}\",\"author_short\":[\"Sunde,  M.\",\"McGrath,  K.\",\"Young,  L.\",\"Matthews,  J.\",\"Chua,  E.\",\"Mackay,  J.\",\"Death,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"sunde-mcgrath-young-matthews-chua-mackay-death-tc1isanoveltumorigenicandnativelydisorderedproteinassociatedwiththyroidcancer-2004\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The C-terminal Domain of Eos Forms a High Order Complex in Solution\",\"type\":\"article\",\"year\":\"2003\",\"pages\":\"42419-42426\",\"volume\":\"278\",\"id\":\"d44e19e1-2ac4-30ab-88a3-a1966922ed83\",\"created\":\"2023-01-10T01:46:13.722Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:13.722Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Ikaros family transcription factors play important roles in the control of hematopoiesis. Family members are predicted to contain up to six classic zinc fingers that are arranged into N- and C-terminal domains. The N-terminal domain is responsible for site-specific DNA binding, whereas the C-terminal domain primarily mediates the homo- and hetero-oligomerization between family members. Although the mechanisms of action of these proteins are not completely understood, the zinc finger domains are known to play a central role. In the current study, we have sought to understand the physical and functional properties of these domains, in particular the C-terminal domain. We show that the N-terminal domain from Eos, and not its C-terminal region, is required to recognize GGGA consensus sequences. Surprisingly, in contrast to the behavior exhibited by Ikaros, the C-terminal domain of Eos inhibits the DNA-binding activity of the full-length protein. In addition, we have used a range of biophysical techniques to demonstrate that the C-terminal domain of Eos mediates the formation of complexes that consist of nine or ten molecules. These results constitute the first direct demonstration that Ikaros family proteins can form higher order complexes in solution, and we discuss this unexpected result in the context of what is currently known about the family members and their possible mechanism of action.\",\"bibtype\":\"article\",\"author\":\"Westman, B.J. and Perdomo, J. and Sunde, M. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M306817200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"43\",\"bibtex\":\"@article{\\n title = {The C-terminal Domain of Eos Forms a High Order Complex in Solution},\\n type = {article},\\n year = {2003},\\n pages = {42419-42426},\\n volume = {278},\\n id = {d44e19e1-2ac4-30ab-88a3-a1966922ed83},\\n created = {2023-01-10T01:46:13.722Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:13.722Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Ikaros family transcription factors play important roles in the control of hematopoiesis. Family members are predicted to contain up to six classic zinc fingers that are arranged into N- and C-terminal domains. The N-terminal domain is responsible for site-specific DNA binding, whereas the C-terminal domain primarily mediates the homo- and hetero-oligomerization between family members. Although the mechanisms of action of these proteins are not completely understood, the zinc finger domains are known to play a central role. In the current study, we have sought to understand the physical and functional properties of these domains, in particular the C-terminal domain. We show that the N-terminal domain from Eos, and not its C-terminal region, is required to recognize GGGA consensus sequences. Surprisingly, in contrast to the behavior exhibited by Ikaros, the C-terminal domain of Eos inhibits the DNA-binding activity of the full-length protein. In addition, we have used a range of biophysical techniques to demonstrate that the C-terminal domain of Eos mediates the formation of complexes that consist of nine or ten molecules. These results constitute the first direct demonstration that Ikaros family proteins can form higher order complexes in solution, and we discuss this unexpected result in the context of what is currently known about the family members and their possible mechanism of action.},\\n bibtype = {article},\\n author = {Westman, B.J. and Perdomo, J. and Sunde, M. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M306817200},\\n journal = {Journal of Biological Chemistry},\\n number = {43}\\n}\",\"author_short\":[\"Westman,  B.\",\"Perdomo,  J.\",\"Sunde,  M.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"westman-perdomo-sunde-crossley-mackay-thecterminaldomainofeosformsahighordercomplexinsolution-2003\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Pentaprobe: a comprehensive sequence for the one-step detection of DNA-binding activities.\",\"type\":\"article\",\"year\":\"2003\",\"volume\":\"31\",\"id\":\"82c033ec-4b64-301f-b28b-440885f5251a\",\"created\":\"2023-01-10T01:46:14.623Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:14.623Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The rapid increase in the number of novel proteins identified in genome projects necessitates simple and rapid methods for assigning function. We describe a strategy for determining whether novel proteins possess typical sequence-specific DNA-binding activity. Many proteins bind recognition sequences of 5 bp or less. Given that there are 4(5) possible 5 bp sites, one might expect the length of sequence required to cover all possibilities would be 4(5) x 5 or 5120 nt. But by allowing overlaps, utilising both strands and using a computer algorithm to generate the minimum sequence, we find the length required is only 516 base pairs. We generated this sequence as six overlapping double-stranded oligonucleotides, termed pentaprobe, and used it in gel retardation experiments to assess DNA binding by both known and putative DNA-binding proteins from several protein families. We have confirmed binding by the zinc finger proteins BKLF, Eos and Pegasus, the Ets domain protein PU.1 and the treble clef N- and C-terminal fingers of GATA-1. We also showed that the N-terminal zinc finger domain of FOG-1 does not behave as a typical DNA-binding domain. Our results suggest that pentaprobe, and related sequences such as hexaprobe, represent useful tools for probing protein function.\",\"bibtype\":\"article\",\"author\":\"Kwan, A.H. and Czolij, R. and Mackay, J.P. and Crossley, M.\",\"doi\":\"10.1093/nar/gng124\",\"journal\":\"Nucleic acids research\",\"number\":\"20\",\"bibtex\":\"@article{\\n title = {Pentaprobe: a comprehensive sequence for the one-step detection of DNA-binding activities.},\\n type = {article},\\n year = {2003},\\n volume = {31},\\n id = {82c033ec-4b64-301f-b28b-440885f5251a},\\n created = {2023-01-10T01:46:14.623Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:14.623Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The rapid increase in the number of novel proteins identified in genome projects necessitates simple and rapid methods for assigning function. We describe a strategy for determining whether novel proteins possess typical sequence-specific DNA-binding activity. Many proteins bind recognition sequences of 5 bp or less. Given that there are 4(5) possible 5 bp sites, one might expect the length of sequence required to cover all possibilities would be 4(5) x 5 or 5120 nt. But by allowing overlaps, utilising both strands and using a computer algorithm to generate the minimum sequence, we find the length required is only 516 base pairs. We generated this sequence as six overlapping double-stranded oligonucleotides, termed pentaprobe, and used it in gel retardation experiments to assess DNA binding by both known and putative DNA-binding proteins from several protein families. We have confirmed binding by the zinc finger proteins BKLF, Eos and Pegasus, the Ets domain protein PU.1 and the treble clef N- and C-terminal fingers of GATA-1. We also showed that the N-terminal zinc finger domain of FOG-1 does not behave as a typical DNA-binding domain. Our results suggest that pentaprobe, and related sequences such as hexaprobe, represent useful tools for probing protein function.},\\n bibtype = {article},\\n author = {Kwan, A.H. and Czolij, R. and Mackay, J.P. and Crossley, M.},\\n doi = {10.1093/nar/gng124},\\n journal = {Nucleic acids research},\\n number = {20}\\n}\",\"author_short\":[\"Kwan,  A.\",\"Czolij,  R.\",\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kwan-czolij-mackay-crossley-pentaprobeacomprehensivesequencefortheonestepdetectionofdnabindingactivities-2003\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The structure of the zinc finger domain from human splicing factor ZNF265 fold\",\"type\":\"article\",\"year\":\"2003\",\"pages\":\"22805-22811\",\"volume\":\"278\",\"id\":\"762d5635-24ff-39d3-85a7-d0f832df931b\",\"created\":\"2023-01-10T01:46:15.539Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:15.539Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Identification of the protein domains that are responsible for RNA recognition has lagged behind the characterization of protein-DNA interactions. However, it is now becoming clear that a range of structural motifs bind to RNA and their structures and molecular mechanisms of action are beginning to be elucidated. In this report, we have expressed and purified one of the two putative RNA-binding domains from ZNF265, a protein that has been shown to bind to the spliceosomal components U1-70K and U2AF35 and to direct alternative splicing. We show that this domain, which contains four highly conserved cysteine residues, forms a stable, monomeric structure upon the addition of 1 molar eq of Zn(II). Determination of the solution structure of this domain reveals a conformation comprising two stacked β-hairpins oriented at ·80° to each other and sandwiching the zinc ion; the fold resembles the zinc ribbon class of zinc-binding domains, although with one less β-strand than most members of the class. Analysis of the structure reveals a striking resemblance to known RNA-binding motifs in terms of the distribution of key surface residues responsible for making RNA contacts, despite a complete lack of structural homology. Furthermore, we have used an RNA gel shift assay to demonstrate that a single crossed finger domain from ZNF265 is capable of binding to an RNA message. Taken together, these results define a new RNA-binding motif and should provide insight into the functions of the >100 uncharacterized proteins in the sequence data bases that contain this domain.\",\"bibtype\":\"article\",\"author\":\"Plambeck, C.A. and Kwan, A.H.Y. and Adams, D.J. and Westman, B.J. and Van der Weyden, L. and Medcalf, R.L. and Morris, B.J. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M301896200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"25\",\"bibtex\":\"@article{\\n title = {The structure of the zinc finger domain from human splicing factor ZNF265 fold},\\n type = {article},\\n year = {2003},\\n pages = {22805-22811},\\n volume = {278},\\n id = {762d5635-24ff-39d3-85a7-d0f832df931b},\\n created = {2023-01-10T01:46:15.539Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:15.539Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Identification of the protein domains that are responsible for RNA recognition has lagged behind the characterization of protein-DNA interactions. However, it is now becoming clear that a range of structural motifs bind to RNA and their structures and molecular mechanisms of action are beginning to be elucidated. In this report, we have expressed and purified one of the two putative RNA-binding domains from ZNF265, a protein that has been shown to bind to the spliceosomal components U1-70K and U2AF35 and to direct alternative splicing. We show that this domain, which contains four highly conserved cysteine residues, forms a stable, monomeric structure upon the addition of 1 molar eq of Zn(II). Determination of the solution structure of this domain reveals a conformation comprising two stacked β-hairpins oriented at ·80° to each other and sandwiching the zinc ion; the fold resembles the zinc ribbon class of zinc-binding domains, although with one less β-strand than most members of the class. Analysis of the structure reveals a striking resemblance to known RNA-binding motifs in terms of the distribution of key surface residues responsible for making RNA contacts, despite a complete lack of structural homology. Furthermore, we have used an RNA gel shift assay to demonstrate that a single crossed finger domain from ZNF265 is capable of binding to an RNA message. Taken together, these results define a new RNA-binding motif and should provide insight into the functions of the >100 uncharacterized proteins in the sequence data bases that contain this domain.},\\n bibtype = {article},\\n author = {Plambeck, C.A. and Kwan, A.H.Y. and Adams, D.J. and Westman, B.J. and Van der Weyden, L. and Medcalf, R.L. and Morris, B.J. and Mackay, J.P.},\\n doi = {10.1074/jbc.M301896200},\\n journal = {Journal of Biological Chemistry},\\n number = {25}\\n}\",\"author_short\":[\"Plambeck,  C.\",\"Kwan,  A.\",\"Adams,  D.\",\"Westman,  B.\",\"Van der Weyden,  L.\",\"Medcalf,  R.\",\"Morris,  B.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"plambeck-kwan-adams-westman-vanderweyden-medcalf-morris-mackay-thestructureofthezincfingerdomainfromhumansplicingfactorznf265fold-2003\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Structural basis for the recognition of Idb1 by the N-terminal LIM domains of LMO2 and LMO4\",\"type\":\"article\",\"year\":\"2003\",\"pages\":\"2224-2233\",\"volume\":\"22\",\"id\":\"443069ef-b4a5-37f0-958b-48eb15aacddd\",\"created\":\"2023-01-10T01:46:16.462Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:16.462Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"LMO2 and LMO4 are members of a small family of nuclear transcriptional regulators that are important for both normal development and disease processes. LMO2 is essential for hemopoiesis and angiogenesis, and inappropriate overexpression of this protein leads to T-cell leukemias. LMO4 is developmentally regulated in the mammary gland and has been implicated in breast oncogenesis. Both proteins comprise two tandemly repeated LIM domains. LMO2 and LMO4 interact with the ubiquitous nuclear adaptor protein Idb1/NLI/CLIM2, which associates with the LIM domains of LMO and LIM homeodomain proteins via its LIM interaction domain (Idb1-LID). We report the solution structures of two LMO:Idb1 complexes (PDB: 1M3V and 1J20) and show that Idb1-LID binds to the N-terminal LIM domain (LIM1) of LMO2 and LMO4 in an extended conformation, contributing a third strand to a β-hairpin in LIM1 domains. These findings constitute the first molecular definition of LIM-mediated protein-protein interactions and suggest a mechanism by which Idb1 can bind a variety of LIM domains that share low sequence homology.\",\"bibtype\":\"article\",\"author\":\"Deane, J.E. and Mackay, J.P. and Kwan, A.H.Y. and Sum, E.Y.M. and Visvader, J.E. and Matthews, J.M.\",\"doi\":\"10.1093/emboj/cdg196\",\"journal\":\"EMBO Journal\",\"number\":\"9\",\"bibtex\":\"@article{\\n title = {Structural basis for the recognition of Idb1 by the N-terminal LIM domains of LMO2 and LMO4},\\n type = {article},\\n year = {2003},\\n pages = {2224-2233},\\n volume = {22},\\n id = {443069ef-b4a5-37f0-958b-48eb15aacddd},\\n created = {2023-01-10T01:46:16.462Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:16.462Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {LMO2 and LMO4 are members of a small family of nuclear transcriptional regulators that are important for both normal development and disease processes. LMO2 is essential for hemopoiesis and angiogenesis, and inappropriate overexpression of this protein leads to T-cell leukemias. LMO4 is developmentally regulated in the mammary gland and has been implicated in breast oncogenesis. Both proteins comprise two tandemly repeated LIM domains. LMO2 and LMO4 interact with the ubiquitous nuclear adaptor protein Idb1/NLI/CLIM2, which associates with the LIM domains of LMO and LIM homeodomain proteins via its LIM interaction domain (Idb1-LID). We report the solution structures of two LMO:Idb1 complexes (PDB: 1M3V and 1J20) and show that Idb1-LID binds to the N-terminal LIM domain (LIM1) of LMO2 and LMO4 in an extended conformation, contributing a third strand to a β-hairpin in LIM1 domains. These findings constitute the first molecular definition of LIM-mediated protein-protein interactions and suggest a mechanism by which Idb1 can bind a variety of LIM domains that share low sequence homology.},\\n bibtype = {article},\\n author = {Deane, J.E. and Mackay, J.P. and Kwan, A.H.Y. and Sum, E.Y.M. and Visvader, J.E. and Matthews, J.M.},\\n doi = {10.1093/emboj/cdg196},\\n journal = {EMBO Journal},\\n number = {9}\\n}\",\"author_short\":[\"Deane,  J.\",\"Mackay,  J.\",\"Kwan,  A.\",\"Sum,  E.\",\"Visvader,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"deane-mackay-kwan-sum-visvader-matthews-structuralbasisfortherecognitionofidb1bythenterminallimdomainsoflmo2andlmo4-2003\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Engineering a protein scaffold from a PHD finger\",\"type\":\"article\",\"year\":\"2003\",\"pages\":\"803-813\",\"volume\":\"11\",\"id\":\"b8887e04-25ff-3420-9eae-4d046cdc7ab2\",\"created\":\"2023-01-10T01:46:17.361Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:17.361Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The design of proteins with tailored functions remains a relatively elusive goal. Small size, a well-defined structure, and the ability to maintain structural integrity despite multiple mutations are all desirable properties for such designer proteins. Many zinc binding domains fit this description. We determined the structure of a PHD finger from the transcriptional cofactor Mi2β and investigated the suitability of this domain as a scaffold for presenting selected binding functions. The two flexible loops in the structure were mutated extensively by either substitution or expansion, without affecting the overall fold of the domain. A binding site for the corepressor CtBP2 was also grafted onto the domain, creating a new PHD domain that can specifically bind CtBP2 both in vitro and in the context of a eukaryotic cell nucleus. These results represent a step toward designing new regulatory proteins for modulating aberrant gene expression in vivo.\",\"bibtype\":\"article\",\"author\":\"Kwan, A.H.Y. and Gell, D.A. and Verger, A. and Crossley, M. and Matthews, J.M. and Mackay, J.P.\",\"doi\":\"10.1016/S0969-2126(03)00122-9\",\"journal\":\"Structure\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {Engineering a protein scaffold from a PHD finger},\\n type = {article},\\n year = {2003},\\n pages = {803-813},\\n volume = {11},\\n id = {b8887e04-25ff-3420-9eae-4d046cdc7ab2},\\n created = {2023-01-10T01:46:17.361Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:17.361Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The design of proteins with tailored functions remains a relatively elusive goal. Small size, a well-defined structure, and the ability to maintain structural integrity despite multiple mutations are all desirable properties for such designer proteins. Many zinc binding domains fit this description. We determined the structure of a PHD finger from the transcriptional cofactor Mi2β and investigated the suitability of this domain as a scaffold for presenting selected binding functions. The two flexible loops in the structure were mutated extensively by either substitution or expansion, without affecting the overall fold of the domain. A binding site for the corepressor CtBP2 was also grafted onto the domain, creating a new PHD domain that can specifically bind CtBP2 both in vitro and in the context of a eukaryotic cell nucleus. These results represent a step toward designing new regulatory proteins for modulating aberrant gene expression in vivo.},\\n bibtype = {article},\\n author = {Kwan, A.H.Y. and Gell, D.A. and Verger, A. and Crossley, M. and Matthews, J.M. and Mackay, J.P.},\\n doi = {10.1016/S0969-2126(03)00122-9},\\n journal = {Structure},\\n number = {7}\\n}\",\"author_short\":[\"Kwan,  A.\",\"Gell,  D.\",\"Verger,  A.\",\"Crossley,  M.\",\"Matthews,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kwan-gell-verger-crossley-matthews-mackay-engineeringaproteinscaffoldfromaphdfinger-2003\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Cardiac hypertrophy and histone deacetylase-dependent transcriptional repression mediated by the atypical homeodomain protein Hop\",\"type\":\"article\",\"year\":\"2003\",\"pages\":\"863-871\",\"volume\":\"112\",\"id\":\"ea3de67e-e52c-3b2d-8ffd-ffe82aeeec88\",\"created\":\"2023-01-10T01:46:18.276Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:18.276Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Activation of multiple pathways is associated with cardiac hypertrophy and heart failure. Repression of antihypertrophic pathways has rarely been demonstrated to cause cardiac hypertrophy in vivo. Hop is an unusual homeodomain protein that is expressed by embryonic and postnatal cardiac myocytes. Unlike other homeodomain proteins, Hop does not bind DNA. Rather, it modulates cardiac growth and proliferation by inhibiting the transcriptional activity of serum response factor (SRF) in cardiomyocytes. Here we show that Hop can inhibit SRF-dependent transcriptional activation by recruiting histone deacetylase (HDAC) activity and can form a complex that includes HDAC2. Transgenic mice that overexpress Hop develop severe cardiac hypertrophy, cardiac fibrosis, and premature death. A mutant form of Hop, which does not recruit HDAC activity, does not induce hypertrophy. Treatment of Hop transgenic mice with trichostatin A, an HDAC inhibitor, prevents hypertrophy. In addition trichostatin A also attenuates hypertrophy induced by infusion of isoproterenol. Thus, chromatin remodeling and repression of otherwise active transcriptional processes can result in hypertrophy and heart failure, and this process can be blocked with chemical HDAC inhibitors.\",\"bibtype\":\"article\",\"author\":\"Kook, H. and Lepore, J.J. and Gitler, A.D. and Lu, M.M. and Yung, W.W.-M. and Mackay, J. and Zhou, R. and Ferrari, V. and Gruber, P. and Epstein, J.A.\",\"doi\":\"10.1172/JCI19137\",\"journal\":\"Journal of Clinical Investigation\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Cardiac hypertrophy and histone deacetylase-dependent transcriptional repression mediated by the atypical homeodomain protein Hop},\\n type = {article},\\n year = {2003},\\n pages = {863-871},\\n volume = {112},\\n id = {ea3de67e-e52c-3b2d-8ffd-ffe82aeeec88},\\n created = {2023-01-10T01:46:18.276Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:18.276Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Activation of multiple pathways is associated with cardiac hypertrophy and heart failure. Repression of antihypertrophic pathways has rarely been demonstrated to cause cardiac hypertrophy in vivo. Hop is an unusual homeodomain protein that is expressed by embryonic and postnatal cardiac myocytes. Unlike other homeodomain proteins, Hop does not bind DNA. Rather, it modulates cardiac growth and proliferation by inhibiting the transcriptional activity of serum response factor (SRF) in cardiomyocytes. Here we show that Hop can inhibit SRF-dependent transcriptional activation by recruiting histone deacetylase (HDAC) activity and can form a complex that includes HDAC2. Transgenic mice that overexpress Hop develop severe cardiac hypertrophy, cardiac fibrosis, and premature death. A mutant form of Hop, which does not recruit HDAC activity, does not induce hypertrophy. Treatment of Hop transgenic mice with trichostatin A, an HDAC inhibitor, prevents hypertrophy. In addition trichostatin A also attenuates hypertrophy induced by infusion of isoproterenol. Thus, chromatin remodeling and repression of otherwise active transcriptional processes can result in hypertrophy and heart failure, and this process can be blocked with chemical HDAC inhibitors.},\\n bibtype = {article},\\n author = {Kook, H. and Lepore, J.J. and Gitler, A.D. and Lu, M.M. and Yung, W.W.-M. and Mackay, J. and Zhou, R. and Ferrari, V. and Gruber, P. and Epstein, J.A.},\\n doi = {10.1172/JCI19137},\\n journal = {Journal of Clinical Investigation},\\n number = {6}\\n}\",\"author_short\":[\"Kook,  H.\",\"Lepore,  J.\",\"Gitler,  A.\",\"Lu,  M.\",\"Yung,  W.\",\"Mackay,  J.\",\"Zhou,  R.\",\"Ferrari,  V.\",\"Gruber,  P.\",\"Epstein,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kook-lepore-gitler-lu-yung-mackay-zhou-ferrari-etal-cardiachypertrophyandhistonedeacetylasedependenttranscriptionalrepressionmediatedbytheatypicalhomeodomainproteinhop-2003\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"CCHX zinc finger derivatives retain the ability to bind Zn(II) and mediate protein-DNA interactions\",\"type\":\"article\",\"year\":\"2003\",\"pages\":\"28011-28018\",\"volume\":\"278\",\"id\":\"254bb9d7-8c93-373a-9c53-0d7edd74d55b\",\"created\":\"2023-01-10T01:46:19.216Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:19.216Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Classical (CCHH) zinc fingers are among the most common protein domains found in eukaryotes. They function as molecular recognition elements that mediate specific contact with DNA, RNA, or other proteins and are composed of a ββα fold surrounding a single zinc ion that is ligated by two cysteine and two histidine residues. In a number of variant zinc fingers, the final histidine is not conserved, and in other unrelated zinc binding domains, residues such as aspartate can function as zinc ligands. To test whether the final histidine is required for normal folding and the DNA-binding function of classical zinc fingers, we focused on finger 3 of basic Krüppel-like factor. The structure of this domain was determined using NMR spectroscopy and found to constitute a typical classical zinc finger. We generated a panel of substitution mutants at the final histidine in this finger and found that several of the mutants retained some ability to fold in the presence of zinc. Consistent with this result, we showed that mutation of the final histidine had only a modest effect on DNA binding in the context of the full three-finger DNA-binding domain of basic Krüppel-like factor. Further, the zinc binding ability of one of the point mutants was tested and found to be indistinguishable from the wild-type domain. These results suggest that the final zinc chelating histidine is not an essential feature of classical zinc fingers and have implications for zinc finger evolution, regulation, and the design of experiments testing the functional roles of these domains.\",\"bibtype\":\"article\",\"author\":\"Simpson, R.J.Y. and Cram, E.D. and Czolij, R. and Matthews, J.M. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M211146200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"30\",\"bibtex\":\"@article{\\n title = {CCHX zinc finger derivatives retain the ability to bind Zn(II) and mediate protein-DNA interactions},\\n type = {article},\\n year = {2003},\\n pages = {28011-28018},\\n volume = {278},\\n id = {254bb9d7-8c93-373a-9c53-0d7edd74d55b},\\n created = {2023-01-10T01:46:19.216Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:19.216Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Classical (CCHH) zinc fingers are among the most common protein domains found in eukaryotes. They function as molecular recognition elements that mediate specific contact with DNA, RNA, or other proteins and are composed of a ββα fold surrounding a single zinc ion that is ligated by two cysteine and two histidine residues. In a number of variant zinc fingers, the final histidine is not conserved, and in other unrelated zinc binding domains, residues such as aspartate can function as zinc ligands. To test whether the final histidine is required for normal folding and the DNA-binding function of classical zinc fingers, we focused on finger 3 of basic Krüppel-like factor. The structure of this domain was determined using NMR spectroscopy and found to constitute a typical classical zinc finger. We generated a panel of substitution mutants at the final histidine in this finger and found that several of the mutants retained some ability to fold in the presence of zinc. Consistent with this result, we showed that mutation of the final histidine had only a modest effect on DNA binding in the context of the full three-finger DNA-binding domain of basic Krüppel-like factor. Further, the zinc binding ability of one of the point mutants was tested and found to be indistinguishable from the wild-type domain. These results suggest that the final zinc chelating histidine is not an essential feature of classical zinc fingers and have implications for zinc finger evolution, regulation, and the design of experiments testing the functional roles of these domains.},\\n bibtype = {article},\\n author = {Simpson, R.J.Y. and Cram, E.D. and Czolij, R. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M211146200},\\n journal = {Journal of Biological Chemistry},\\n number = {30}\\n}\",\"author_short\":[\"Simpson,  R.\",\"Cram,  E.\",\"Czolij,  R.\",\"Matthews,  J.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"simpson-cram-czolij-matthews-crossley-mackay-cchxzincfingerderivativesretaintheabilitytobindzniiandmediateproteindnainteractions-2003\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution structure and behaviour of Δ-m-α-[Ru(R,R- picchxnMe<inf>2</inf>)(phi)]<sup>2+</sup> by NMR spectroscopy and molecular modelling\",\"type\":\"article\",\"year\":\"2003\",\"pages\":\"165-170\",\"id\":\"a5866cc0-0e9f-38d7-a4df-ab0bef4ad5e5\",\"created\":\"2023-01-10T01:46:20.125Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:20.125Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The self-association of the DNA metalloprobe Δ-cis-α-[Ru(R,R- picchxnMe2)(phi)]2+ (α-phi) in aqueous solution has been investigated using 1H NMR spectroscopy and molecular modelling. The concentration dependence of proton chemical shifts of the complex gave initial indications of a self-associated species, while its structural isomer Δ-cis-β-[Ru(R,R-picchxnMe2)(phi)]2+ (β-phi) showed no such dependence. 2D-COSY and 2D-ROESY experiments were used for the complete assignment of the proton resonances of both isomers and allowed a qualitative determination of the self-association of the a isomer through the detection of intermolecular ROEs. NMR spectroscopy can also be effectively used to differentiate Δ- and Λ-diastereomers. In addition, we show, by pulsed field gradient longitudinal eddy-current delay (PFGLED) NMR spectroscopy, that α-phi self-associates at higher concentrations with an effective molecular weight at 25 mM three times that at 2.5 mM. This apparent oligomerisation was not observed for the β-isomer. © The Royal Society of Chemistry 2003.\",\"bibtype\":\"article\",\"author\":\"Proudfoot, E.M. and Mackay, J.P. and Karuso, P.\",\"doi\":\"10.1039/b208846k\",\"journal\":\"Dalton Transactions\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Solution structure and behaviour of Δ-m-α-[Ru(R,R- picchxnMe<inf>2</inf>)(phi)]<sup>2+</sup> by NMR spectroscopy and molecular modelling},\\n type = {article},\\n year = {2003},\\n pages = {165-170},\\n id = {a5866cc0-0e9f-38d7-a4df-ab0bef4ad5e5},\\n created = {2023-01-10T01:46:20.125Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:20.125Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The self-association of the DNA metalloprobe Δ-cis-α-[Ru(R,R- picchxnMe2)(phi)]2+ (α-phi) in aqueous solution has been investigated using 1H NMR spectroscopy and molecular modelling. The concentration dependence of proton chemical shifts of the complex gave initial indications of a self-associated species, while its structural isomer Δ-cis-β-[Ru(R,R-picchxnMe2)(phi)]2+ (β-phi) showed no such dependence. 2D-COSY and 2D-ROESY experiments were used for the complete assignment of the proton resonances of both isomers and allowed a qualitative determination of the self-association of the a isomer through the detection of intermolecular ROEs. NMR spectroscopy can also be effectively used to differentiate Δ- and Λ-diastereomers. In addition, we show, by pulsed field gradient longitudinal eddy-current delay (PFGLED) NMR spectroscopy, that α-phi self-associates at higher concentrations with an effective molecular weight at 25 mM three times that at 2.5 mM. This apparent oligomerisation was not observed for the β-isomer. © The Royal Society of Chemistry 2003.},\\n bibtype = {article},\\n author = {Proudfoot, E.M. and Mackay, J.P. and Karuso, P.},\\n doi = {10.1039/b208846k},\\n journal = {Dalton Transactions},\\n number = {2}\\n}\",\"author_short\":[\"Proudfoot,  E.\",\"Mackay,  J.\",\"Karuso,  P.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"proudfoot-mackay-karuso-solutionstructureandbehaviourofmrurrpicchxnmeinf2infphisup2supbynmrspectroscopyandmolecularmodelling-2003\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Biophysical characterization of the α-globin binding protein α-hemoglobin stabilizing protein\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"40602-40609\",\"volume\":\"277\",\"id\":\"7bd08e1a-428f-320d-b146-14956285c9d5\",\"created\":\"2023-01-10T01:46:21.028Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:21.028Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"α-Hemoglobin stabilizing protein (AHSP) is a small (12 kDa) and abundant erythroid-specific protein that binds specifically to free α-(hemo)globin and prevents its precipitation. When present in excess over β-globin, its normal binding partner, α-globin can have severe cytotoxic effects that contribute to important human diseases such as β-thalassemia. Because AHSP might act as a chaperone to prevent the harmful aggregation of α-globin during normal erythroid cell development and in diseases of globin chain imbalance, it is important to characterize the biochemical properties of the AHSP·α-globin complex. Here we provide the first structural information about AHSP and its interaction with α-globin. We find that AHSP is a predominantly α-helical globular protein with a somewhat asymmetric shape. AHSP and α-globin are both monomeric in solution as determined by analytical ultracentrifugation and bind each other to form a complex with 1:1 subunit stoichiometry, as judged by gel filtration and amino acid analysis. We have used isothermal titration calorimetry to show that the interaction is of moderate affinity with an association constant of 1 × 107 M-1 and is thus likely to be biologically significant given the concentration of AHSP (∼0.1 mM) and hemoglobin (∼4 mm) in the late pro-erythroblast.\",\"bibtype\":\"article\",\"author\":\"Dvid, G. and Yi, K. and Eaton A, S.A. and Weiss C, M.J. and Mackay E, J.P.\",\"doi\":\"10.1074/jbc.M206084200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"43\",\"bibtex\":\"@article{\\n title = {Biophysical characterization of the α-globin binding protein α-hemoglobin stabilizing protein},\\n type = {article},\\n year = {2002},\\n pages = {40602-40609},\\n volume = {277},\\n id = {7bd08e1a-428f-320d-b146-14956285c9d5},\\n created = {2023-01-10T01:46:21.028Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:21.028Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {α-Hemoglobin stabilizing protein (AHSP) is a small (12 kDa) and abundant erythroid-specific protein that binds specifically to free α-(hemo)globin and prevents its precipitation. When present in excess over β-globin, its normal binding partner, α-globin can have severe cytotoxic effects that contribute to important human diseases such as β-thalassemia. Because AHSP might act as a chaperone to prevent the harmful aggregation of α-globin during normal erythroid cell development and in diseases of globin chain imbalance, it is important to characterize the biochemical properties of the AHSP·α-globin complex. Here we provide the first structural information about AHSP and its interaction with α-globin. We find that AHSP is a predominantly α-helical globular protein with a somewhat asymmetric shape. AHSP and α-globin are both monomeric in solution as determined by analytical ultracentrifugation and bind each other to form a complex with 1:1 subunit stoichiometry, as judged by gel filtration and amino acid analysis. We have used isothermal titration calorimetry to show that the interaction is of moderate affinity with an association constant of 1 × 107 M-1 and is thus likely to be biologically significant given the concentration of AHSP (∼0.1 mM) and hemoglobin (∼4 mm) in the late pro-erythroblast.},\\n bibtype = {article},\\n author = {Dvid, G. and Yi, K. and Eaton A, S.A. and Weiss C, M.J. and Mackay E, J.P.},\\n doi = {10.1074/jbc.M206084200},\\n journal = {Journal of Biological Chemistry},\\n number = {43}\\n}\",\"author_short\":[\"Dvid,  G.\",\"Yi,  K.\",\"Eaton A,  S.\",\"Weiss C,  M.\",\"Mackay E,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dvid-yi-eatona-weissc-mackaye-biophysicalcharacterizationoftheglobinbindingproteinhemoglobinstabilizingprotein-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"A new zinc binding fold underlines the versatility of zinc binding modules in protein evolution\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"639-648\",\"volume\":\"10\",\"id\":\"3d10c7ef-8426-3256-9aa7-a918db603a09\",\"created\":\"2023-01-10T01:46:21.952Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:21.952Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Many different zinc binding modules have been identified. Their abundance and variety suggests that the formation of zinc binding folds might be relatively common. We have determined the structure of CH11, a 27-residue peptide derived from the first cysteine/histidine-rich region (CH1) of CREB binding protein (CBP). This peptide forms a highly ordered zinc-dependent fold that is distinct from known folds. The structure differs from a subsequently determined structure of a larger region from the CH3 region of CBP, and the CH11 fold probably represents a nonphysiologically active form. Despite this, the fold is thermostable and tolerant to both multiple alanine mutations and changes in the zinc-ligand spacing. Our data support the idea that zinc binding domains may arise frequently. Additionally, such structures may prove useful as scaffolds for protein design, given their stability and robustness.\",\"bibtype\":\"article\",\"author\":\"Sharpe, B.K. and Matthews, J.M. and Kwan, A.H.Y. and Newton, A. and Gell, D.A. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1016/S0969-2126(02)00757-8\",\"journal\":\"Structure\",\"number\":\"5\",\"bibtex\":\"@article{\\n title = {A new zinc binding fold underlines the versatility of zinc binding modules in protein evolution},\\n type = {article},\\n year = {2002},\\n pages = {639-648},\\n volume = {10},\\n id = {3d10c7ef-8426-3256-9aa7-a918db603a09},\\n created = {2023-01-10T01:46:21.952Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:21.952Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Many different zinc binding modules have been identified. Their abundance and variety suggests that the formation of zinc binding folds might be relatively common. We have determined the structure of CH11, a 27-residue peptide derived from the first cysteine/histidine-rich region (CH1) of CREB binding protein (CBP). This peptide forms a highly ordered zinc-dependent fold that is distinct from known folds. The structure differs from a subsequently determined structure of a larger region from the CH3 region of CBP, and the CH11 fold probably represents a nonphysiologically active form. Despite this, the fold is thermostable and tolerant to both multiple alanine mutations and changes in the zinc-ligand spacing. Our data support the idea that zinc binding domains may arise frequently. Additionally, such structures may prove useful as scaffolds for protein design, given their stability and robustness.},\\n bibtype = {article},\\n author = {Sharpe, B.K. and Matthews, J.M. and Kwan, A.H.Y. and Newton, A. and Gell, D.A. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1016/S0969-2126(02)00757-8},\\n journal = {Structure},\\n number = {5}\\n}\",\"author_short\":[\"Sharpe,  B.\",\"Matthews,  J.\",\"Kwan,  A.\",\"Newton,  A.\",\"Gell,  D.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"sharpe-matthews-kwan-newton-gell-crossley-mackay-anewzincbindingfoldunderlinestheversatilityofzincbindingmodulesinproteinevolution-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"kwan,  a\":\"https://www.kwanlabusyd.com/publications-1\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of FLIN4, an intramolecular LMO4:ldb1 complex [8]\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"165-166\",\"volume\":\"23\",\"id\":\"d2aa00f0-07ce-3979-9e30-49c883eec666\",\"created\":\"2023-01-10T01:46:22.869Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:22.869Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Deane, J.E. and Visvader, J.E. and Mackay, J.P. and Matthews, J.M.\",\"doi\":\"10.1023/A:1016363414644\",\"journal\":\"Journal of Biomolecular NMR\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of FLIN4, an intramolecular LMO4:ldb1 complex [8]},\\n type = {article},\\n year = {2002},\\n pages = {165-166},\\n volume = {23},\\n id = {d2aa00f0-07ce-3979-9e30-49c883eec666},\\n created = {2023-01-10T01:46:22.869Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:22.869Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Deane, J.E. and Visvader, J.E. and Mackay, J.P. and Matthews, J.M.},\\n doi = {10.1023/A:1016363414644},\\n journal = {Journal of Biomolecular NMR},\\n number = {2}\\n}\",\"author_short\":[\"Deane,  J.\",\"Visvader,  J.\",\"Mackay,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"deane-visvader-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofflin4anintramolecularlmo4ldb1complex8-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution structure of a hydrophobic analogue of the winter flounder antifreeze protein\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"1259-1266\",\"volume\":\"269\",\"id\":\"e7419261-1955-324a-9050-96596e299897\",\"created\":\"2023-01-10T01:46:23.798Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:23.798Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The solution structure of a synthetic mutant type I antifreeze protein (AFP I) was determined in aqueous solution at pH 7.0 using nuclear magnetic resonance (NMR) spectroscopy. The mutations comprised the replacement of the four Thr residues by Val and the introduction of two additional Lys-Glu salt bridges. The antifreeze activity of this mutant peptide, VVVV2KE, has been previously shown to be similar to that of the wild type protein, HPLC6 (defined here as TTTT). The solution structure reveals an ahelix bent in the same direction as the more bent conformer of the published crystalstructure of TTTT, while the side chain χ1 rotamers of VVVV2KE are similar to those of the straighter conformer in the crystal of TTTT. The Val side chains of VVVV2KE assume the same orientations as the Thr side chains of TTTT, confirming the conservative nature of this mutation. The combined data suggest that AFP I undergoes an equilibrium between straight and bent helices in solution, combined with independent equilibria between different side chain rotamers for some of the amino acid residues. The present study presents the first complete sequence-specific resonance assignments and the first complete solution structure determination by NMR of any AFP I protein.\",\"bibtype\":\"article\",\"author\":\"Liepinsh, E. and Otting, G. and Harding, M.M. and Ward, L.G. and Mackay, J.P. and Haymet, A.D.J.\",\"doi\":\"10.1046/j.1432-1033.2002.02766.x\",\"journal\":\"European Journal of Biochemistry\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {Solution structure of a hydrophobic analogue of the winter flounder antifreeze protein},\\n type = {article},\\n year = {2002},\\n pages = {1259-1266},\\n volume = {269},\\n id = {e7419261-1955-324a-9050-96596e299897},\\n created = {2023-01-10T01:46:23.798Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:23.798Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The solution structure of a synthetic mutant type I antifreeze protein (AFP I) was determined in aqueous solution at pH 7.0 using nuclear magnetic resonance (NMR) spectroscopy. The mutations comprised the replacement of the four Thr residues by Val and the introduction of two additional Lys-Glu salt bridges. The antifreeze activity of this mutant peptide, VVVV2KE, has been previously shown to be similar to that of the wild type protein, HPLC6 (defined here as TTTT). The solution structure reveals an ahelix bent in the same direction as the more bent conformer of the published crystalstructure of TTTT, while the side chain χ1 rotamers of VVVV2KE are similar to those of the straighter conformer in the crystal of TTTT. The Val side chains of VVVV2KE assume the same orientations as the Thr side chains of TTTT, confirming the conservative nature of this mutation. The combined data suggest that AFP I undergoes an equilibrium between straight and bent helices in solution, combined with independent equilibria between different side chain rotamers for some of the amino acid residues. The present study presents the first complete sequence-specific resonance assignments and the first complete solution structure determination by NMR of any AFP I protein.},\\n bibtype = {article},\\n author = {Liepinsh, E. and Otting, G. and Harding, M.M. and Ward, L.G. and Mackay, J.P. and Haymet, A.D.J.},\\n doi = {10.1046/j.1432-1033.2002.02766.x},\\n journal = {European Journal of Biochemistry},\\n number = {4}\\n}\",\"author_short\":[\"Liepinsh,  E.\",\"Otting,  G.\",\"Harding,  M.\",\"Ward,  L.\",\"Mackay,  J.\",\"Haymet,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"liepinsh-otting-harding-ward-mackay-haymet-solutionstructureofahydrophobicanalogueofthewinterflounderantifreezeprotein-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Ikaros: A key regulator of haematopoiesis\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"1304-1307\",\"volume\":\"34\",\"id\":\"b11bff38-f90a-3ab1-9e2f-59aa100f8266\",\"created\":\"2023-01-10T01:46:24.702Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:24.702Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function - unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription. © 2002 Elsevier Science Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Westman, B.J. and Mackay, J.P. and Gell, D.\",\"doi\":\"10.1016/S1357-2725(02)00070-5\",\"journal\":\"International Journal of Biochemistry and Cell Biology\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {Ikaros: A key regulator of haematopoiesis},\\n type = {article},\\n year = {2002},\\n pages = {1304-1307},\\n volume = {34},\\n id = {b11bff38-f90a-3ab1-9e2f-59aa100f8266},\\n created = {2023-01-10T01:46:24.702Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:24.702Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function - unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription. © 2002 Elsevier Science Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Westman, B.J. and Mackay, J.P. and Gell, D.},\\n doi = {10.1016/S1357-2725(02)00070-5},\\n journal = {International Journal of Biochemistry and Cell Biology},\\n number = {10}\\n}\",\"author_short\":[\"Westman,  B.\",\"Mackay,  J.\",\"Gell,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Characterization of the conserved interaction between GATA and FOG family proteins\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"35720-35729\",\"volume\":\"277\",\"id\":\"d4514158-aef2-3645-95c3-a537e76c54ce\",\"created\":\"2023-01-10T01:46:25.606Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:25.606Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.\",\"bibtype\":\"article\",\"author\":\"Kowalski, K. and Liew, C.K. and Matthews, J.M. and Gell, D.A. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M204663200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"38\",\"bibtex\":\"@article{\\n title = {Characterization of the conserved interaction between GATA and FOG family proteins},\\n type = {article},\\n year = {2002},\\n pages = {35720-35729},\\n volume = {277},\\n id = {d4514158-aef2-3645-95c3-a537e76c54ce},\\n created = {2023-01-10T01:46:25.606Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:25.606Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.},\\n bibtype = {article},\\n author = {Kowalski, K. and Liew, C.K. and Matthews, J.M. and Gell, D.A. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M204663200},\\n journal = {Journal of Biological Chemistry},\\n number = {38}\\n}\",\"author_short\":[\"Kowalski,  K.\",\"Liew,  C.\",\"Matthews,  J.\",\"Gell,  D.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Characterization of the conserved interaction between GATA and FOG family proteins\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"35720-35729\",\"volume\":\"277\",\"id\":\"943dca0a-527a-3b78-8975-163e0bc0a3c9\",\"created\":\"2023-01-10T01:46:26.825Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:26.825Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.\",\"bibtype\":\"article\",\"author\":\"Kowalski, K. and Liew, C.K. and Matthews, J.M. and Gell, D.A. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M204663200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"38\",\"bibtex\":\"@article{\\n title = {Characterization of the conserved interaction between GATA and FOG family proteins},\\n type = {article},\\n year = {2002},\\n pages = {35720-35729},\\n volume = {277},\\n id = {943dca0a-527a-3b78-8975-163e0bc0a3c9},\\n created = {2023-01-10T01:46:26.825Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:26.825Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.},\\n bibtype = {article},\\n author = {Kowalski, K. and Liew, C.K. and Matthews, J.M. and Gell, D.A. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M204663200},\\n journal = {Journal of Biological Chemistry},\\n number = {38}\\n}\",\"author_short\":[\"Kowalski,  K.\",\"Liew,  C.\",\"Matthews,  J.\",\"Gell,  D.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The Bacillus subtilis cell division proteins FtsL and DivIC are intrinsically unstable and do not interact with one another in the absence of other septasomal components\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"663-674\",\"volume\":\"44\",\"id\":\"8a1d4881-9023-34c3-97a4-9fab972fd4c2\",\"created\":\"2023-01-10T01:46:27.729Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:27.729Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The bacterial septum appears to comprise a macromolecular assembly of essential cell division proteins (the 'septasome') that are responsible for physically dividing the cell during cytokinesis. FtsL and DivIC are essential components of this division machinery in Bacillus subtilis. We used yeast two-hybrid analysis as well as a variety of biochemical and biophysical methods to examine the proposed interaction between Bacillus subtilis FtsL and DivIC. We show that FtsL and DivIC are thermodynamically unstable proteins that are likely to be unfolded and therefore targeted for degradation unless stabilized by interactions with other components of the septasome. However, we show that this stabilization does not result from a direct interaction between FtsL and DivIC. We propose that the observed interdependence of DivIC and FtsL stability is a result of indirect interactions that are mediated by other septasomal proteins.\",\"bibtype\":\"article\",\"author\":\"Robson, S.A. and Michie, K.A. and Mackay, J.P. and Harry, E. and King, G.F.\",\"doi\":\"10.1046/j.1365-2958.2002.02920.x\",\"journal\":\"Molecular Microbiology\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {The Bacillus subtilis cell division proteins FtsL and DivIC are intrinsically unstable and do not interact with one another in the absence of other septasomal components},\\n type = {article},\\n year = {2002},\\n pages = {663-674},\\n volume = {44},\\n id = {8a1d4881-9023-34c3-97a4-9fab972fd4c2},\\n created = {2023-01-10T01:46:27.729Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:27.729Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The bacterial septum appears to comprise a macromolecular assembly of essential cell division proteins (the 'septasome') that are responsible for physically dividing the cell during cytokinesis. FtsL and DivIC are essential components of this division machinery in Bacillus subtilis. We used yeast two-hybrid analysis as well as a variety of biochemical and biophysical methods to examine the proposed interaction between Bacillus subtilis FtsL and DivIC. We show that FtsL and DivIC are thermodynamically unstable proteins that are likely to be unfolded and therefore targeted for degradation unless stabilized by interactions with other components of the septasome. However, we show that this stabilization does not result from a direct interaction between FtsL and DivIC. We propose that the observed interdependence of DivIC and FtsL stability is a result of indirect interactions that are mediated by other septasomal proteins.},\\n bibtype = {article},\\n author = {Robson, S.A. and Michie, K.A. and Mackay, J.P. and Harry, E. and King, G.F.},\\n doi = {10.1046/j.1365-2958.2002.02920.x},\\n journal = {Molecular Microbiology},\\n number = {3}\\n}\",\"author_short\":[\"Robson,  S.\",\"Michie,  K.\",\"Mackay,  J.\",\"Harry,  E.\",\"King,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"robson-michie-mackay-harry-king-thebacillussubtiliscelldivisionproteinsftslanddivicareintrinsicallyunstableanddonotinteractwithoneanotherintheabsenceofotherseptasomalcomponents-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Hop is an unusual homeobox gene that modulates cardiac development\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"713-723\",\"volume\":\"110\",\"id\":\"31c23e13-969a-3713-9fa5-129a498c180a\",\"created\":\"2023-01-10T01:46:28.648Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:28.648Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Hop is a small, divergent homeodomain protein that lacks certain conserved residues required for DNA binding. Hop gene expression initiates early in cardiogenesis and continues in cardiomyocytes throughout embryonic and postnatal development. Genetic and biochemical data indicate that Hop functions directly downstream of Nkx2-5. Inactivation of Hop in mice by homologous recombination results in a partially penetrant embryonic lethal phenotype with severe developmental cardiac defects involving the myocardium. Inhibition of Hop activity in zebrafish embryos likewise disrupts cardiac development and results in severely impaired cardiac function. Hop physically interacts with serum response factor (SRF) and inhibits activation of SRF-dependent transcription by inhibiting SRF binding to DNA. Hop encodes an unusual homeodomain protein that modulates SRF-dependent cardiac-specific gene expression and cardiac development.\",\"bibtype\":\"article\",\"author\":\"Chen, F. and Kook, H. and Milewski, R. and Gitler, A.D. and Lu, M.M. and Li, J. and Nazarian, R. and Schnepp, R. and Jen, K. and Biben, C. and Mullins, M.C. and Epstein, J.A.\",\"doi\":\"10.1016/S0092-8674(02)00932-7\",\"journal\":\"Cell\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Hop is an unusual homeobox gene that modulates cardiac development},\\n type = {article},\\n year = {2002},\\n pages = {713-723},\\n volume = {110},\\n id = {31c23e13-969a-3713-9fa5-129a498c180a},\\n created = {2023-01-10T01:46:28.648Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:28.648Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Hop is a small, divergent homeodomain protein that lacks certain conserved residues required for DNA binding. Hop gene expression initiates early in cardiogenesis and continues in cardiomyocytes throughout embryonic and postnatal development. Genetic and biochemical data indicate that Hop functions directly downstream of Nkx2-5. Inactivation of Hop in mice by homologous recombination results in a partially penetrant embryonic lethal phenotype with severe developmental cardiac defects involving the myocardium. Inhibition of Hop activity in zebrafish embryos likewise disrupts cardiac development and results in severely impaired cardiac function. Hop physically interacts with serum response factor (SRF) and inhibits activation of SRF-dependent transcription by inhibiting SRF binding to DNA. Hop encodes an unusual homeodomain protein that modulates SRF-dependent cardiac-specific gene expression and cardiac development.},\\n bibtype = {article},\\n author = {Chen, F. and Kook, H. and Milewski, R. and Gitler, A.D. and Lu, M.M. and Li, J. and Nazarian, R. and Schnepp, R. and Jen, K. and Biben, C. and Mullins, M.C. and Epstein, J.A.},\\n doi = {10.1016/S0092-8674(02)00932-7},\\n journal = {Cell},\\n number = {6}\\n}\",\"author_short\":[\"Chen,  F.\",\"Kook,  H.\",\"Milewski,  R.\",\"Gitler,  A.\",\"Lu,  M.\",\"Li,  J.\",\"Nazarian,  R.\",\"Schnepp,  R.\",\"Jen,  K.\",\"Biben,  C.\",\"Mullins,  M.\",\"Epstein,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"chen-kook-milewski-gitler-lu-li-nazarian-schnepp-etal-hopisanunusualhomeoboxgenethatmodulatescardiacdevelopment-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Siah ubiquitin ligase is structurally related to traf and modulates tnf-α signaling\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"68-75\",\"volume\":\"9\",\"id\":\"4c060996-79de-3f5c-ae4a-e1f05aebf808\",\"created\":\"2023-01-10T01:46:29.556Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:29.556Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Members of the Siah (seven in absentia homolog) family of RING domain proteins are components of E3 ubiquitin ligase complexes that catalyze ubiquitination of proteins. We have determined the crystal structure of the substrate-binding domain (SBD) of murine Siah1a to 2.6 Å resolution. The structure reveals that Siah is a dimeric protein and that the SBD adopts an eight-stranded β-sandwich fold that is highly similar to the TRAF-C region of TRAF (TNF-receptor associated factor) proteins. The TRAF-C region interacts with TNF-α receptors and TNF-receptor associated death-domain (TRADD) proteins; however, our findings indicate that these interactions are unlikely to be mimicked by Siah. The Siah structure also reveals two novel zinc fingers in a region with sequence similarity to TRAF. We find that the Siah1a SBD potentiates TNF-α-mediated NF-κB activation. Therefore, Siah proteins share important similarities with the TRAF family of proteins, including their overall domain architecture, three-dimensional structure and functional activity. © 2002 Nature Publishing Group.\",\"bibtype\":\"article\",\"author\":\"Polekhina, G. and House, C.M. and Traficante, N. and Mackay, J.P. and Relaix, F. and Sassoon, D.A. and Parker, M.W. and Bowtell, D.D.L.\",\"doi\":\"10.1038/nsb743\",\"journal\":\"Nature Structural Biology\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Siah ubiquitin ligase is structurally related to traf and modulates tnf-α signaling},\\n type = {article},\\n year = {2002},\\n pages = {68-75},\\n volume = {9},\\n id = {4c060996-79de-3f5c-ae4a-e1f05aebf808},\\n created = {2023-01-10T01:46:29.556Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:29.556Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Members of the Siah (seven in absentia homolog) family of RING domain proteins are components of E3 ubiquitin ligase complexes that catalyze ubiquitination of proteins. We have determined the crystal structure of the substrate-binding domain (SBD) of murine Siah1a to 2.6 Å resolution. The structure reveals that Siah is a dimeric protein and that the SBD adopts an eight-stranded β-sandwich fold that is highly similar to the TRAF-C region of TRAF (TNF-receptor associated factor) proteins. The TRAF-C region interacts with TNF-α receptors and TNF-receptor associated death-domain (TRADD) proteins; however, our findings indicate that these interactions are unlikely to be mimicked by Siah. The Siah structure also reveals two novel zinc fingers in a region with sequence similarity to TRAF. We find that the Siah1a SBD potentiates TNF-α-mediated NF-κB activation. Therefore, Siah proteins share important similarities with the TRAF family of proteins, including their overall domain architecture, three-dimensional structure and functional activity. © 2002 Nature Publishing Group.},\\n bibtype = {article},\\n author = {Polekhina, G. and House, C.M. and Traficante, N. and Mackay, J.P. and Relaix, F. and Sassoon, D.A. and Parker, M.W. and Bowtell, D.D.L.},\\n doi = {10.1038/nsb743},\\n journal = {Nature Structural Biology},\\n number = {1}\\n}\",\"author_short\":[\"Polekhina,  G.\",\"House,  C.\",\"Traficante,  N.\",\"Mackay,  J.\",\"Relaix,  F.\",\"Sassoon,  D.\",\"Parker,  M.\",\"Bowtell,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"polekhina-house-traficante-mackay-relaix-sassoon-parker-bowtell-siahubiquitinligaseisstructurallyrelatedtotrafandmodulatestnfsignaling-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Type I shorthorn sculpin antifreeze protein: Recombinant synthesis, solution conformation, and ice growth inhibition studies\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"24073-24080\",\"volume\":\"277\",\"id\":\"3b95bafc-8671-34e7-80d0-daa877523c66\",\"created\":\"2023-01-10T01:46:30.477Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:30.477Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"A number of structurally diverse classes of \\\"antifreeze\\\" proteins that allow fish to survive in sub-zero ice-laden waters have been isolated from the blood plasma of cold water teleosts. However, despite receiving a great deal of attention, the one or more mechanisms through which these proteins act are not fully understood. In this report we have synthesized a type I antifreeze polypeptide (AFP) from the shorthorn sculpin Myoxocephalus scorpius using recombinant methods. Construction of a synthetic gene with optimized codon usage and expression as a glutathione S-transferase fusion protein followed by purification yielded milligram amounts of polypeptide with two extra residues appended to the N terminus. Circular dichroism and NMR experiments, including residual dipolar coupling measurements on a 15N-labeled recombinant polypeptide, show that the polypeptides are α-helical with the first four residues being more flexible than the remainder of the sequence. Both the recombinant and synthetic polypeptides modify ice growth, forming facetted crystals just below the freezing point, but display negligible thermal hysteresis. Acetylation of Lys-10, Lys-20, and Lys-21 as well as the N terminus of the recombinant polypeptide gave a derivative that displays both thermal hysteresis (0.4°C at 15 mg/ml) and ice crystal faceting. These results confirm that the N terminus of wild-type polypeptide is functionally important and support our previously proposed mechanism for all type I proteins, in which the hydrophobic face is oriented toward the ice at the ice/water interface.\",\"bibtype\":\"article\",\"author\":\"Fairley, K. and Westman, B.J. and Pham, L.H. and Haymet, A.D.J. and Harding, M.M. and Mackay, J.P.\",\"doi\":\"10.1074/jbc.M200307200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"27\",\"bibtex\":\"@article{\\n title = {Type I shorthorn sculpin antifreeze protein: Recombinant synthesis, solution conformation, and ice growth inhibition studies},\\n type = {article},\\n year = {2002},\\n pages = {24073-24080},\\n volume = {277},\\n id = {3b95bafc-8671-34e7-80d0-daa877523c66},\\n created = {2023-01-10T01:46:30.477Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:30.477Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {A number of structurally diverse classes of \\\"antifreeze\\\" proteins that allow fish to survive in sub-zero ice-laden waters have been isolated from the blood plasma of cold water teleosts. However, despite receiving a great deal of attention, the one or more mechanisms through which these proteins act are not fully understood. In this report we have synthesized a type I antifreeze polypeptide (AFP) from the shorthorn sculpin Myoxocephalus scorpius using recombinant methods. Construction of a synthetic gene with optimized codon usage and expression as a glutathione S-transferase fusion protein followed by purification yielded milligram amounts of polypeptide with two extra residues appended to the N terminus. Circular dichroism and NMR experiments, including residual dipolar coupling measurements on a 15N-labeled recombinant polypeptide, show that the polypeptides are α-helical with the first four residues being more flexible than the remainder of the sequence. Both the recombinant and synthetic polypeptides modify ice growth, forming facetted crystals just below the freezing point, but display negligible thermal hysteresis. Acetylation of Lys-10, Lys-20, and Lys-21 as well as the N terminus of the recombinant polypeptide gave a derivative that displays both thermal hysteresis (0.4°C at 15 mg/ml) and ice crystal faceting. These results confirm that the N terminus of wild-type polypeptide is functionally important and support our previously proposed mechanism for all type I proteins, in which the hydrophobic face is oriented toward the ice at the ice/water interface.},\\n bibtype = {article},\\n author = {Fairley, K. and Westman, B.J. and Pham, L.H. and Haymet, A.D.J. and Harding, M.M. and Mackay, J.P.},\\n doi = {10.1074/jbc.M200307200},\\n journal = {Journal of Biological Chemistry},\\n number = {27}\\n}\",\"author_short\":[\"Fairley,  K.\",\"Westman,  B.\",\"Pham,  L.\",\"Haymet,  A.\",\"Harding,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"fairley-westman-pham-haymet-harding-mackay-typeishorthornsculpinantifreezeproteinrecombinantsynthesissolutionconformationandicegrowthinhibitionstudies-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The solution structure and intramolecular associations of the Tec kinase Src homology 3 domain\",\"type\":\"article\",\"year\":\"2002\",\"pages\":\"755-762\",\"volume\":\"277\",\"id\":\"b3f30832-cb31-30ae-a155-a163a5517400\",\"created\":\"2023-01-10T01:46:31.373Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:31.373Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Tec is the prototypic member of a family of intracellular tyrosine kinases that includes Txk, Bmx, Itk, and Btk. Tec family kinases share similarities in domain structure with Src family kinases, but one of the features that differentiates them is a proline-rich region (PRR) preceding their Src homology (SH) 3 domain. Evidence that the PRR of Itk can bind in an intramolecular fashion to its SH3 domain and the lack of a regulatory tyrosine in the C terminus indicates that Tec kinases must be regulated by a different set of intramolecular interactions to the Src kinases. We have determined the solution structure of the Tec SH3 domain and have investigated interactions with its PRR, which contains two SH3-binding sites. We demonstrate that in vitro, the Tec PRR can bind in an intramolecular fashion to the SH3. However, the affinity is lower than that for dimerization via reciprocal PRR-SH3 association. Using site-directed mutagenesis we show that both sites can bind the Tec SH3 domain; site 1 (155KTLPPAP161) binds intramolecularly, while site 2 (165KRRPPPPIPP174) cannot and binds in an intermolecular fashion. These distinct roles for the SH3 binding sites in Tec family kinases could be important for protein targeting and enzyme activation.\",\"bibtype\":\"article\",\"author\":\"Pursglove, S.E. and Mulhern, T.D. and Mackay, J.P. and Hinds, M.G. and Booker, G.W.\",\"doi\":\"10.1074/jbc.M108318200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {The solution structure and intramolecular associations of the Tec kinase Src homology 3 domain},\\n type = {article},\\n year = {2002},\\n pages = {755-762},\\n volume = {277},\\n id = {b3f30832-cb31-30ae-a155-a163a5517400},\\n created = {2023-01-10T01:46:31.373Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:31.373Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Tec is the prototypic member of a family of intracellular tyrosine kinases that includes Txk, Bmx, Itk, and Btk. Tec family kinases share similarities in domain structure with Src family kinases, but one of the features that differentiates them is a proline-rich region (PRR) preceding their Src homology (SH) 3 domain. Evidence that the PRR of Itk can bind in an intramolecular fashion to its SH3 domain and the lack of a regulatory tyrosine in the C terminus indicates that Tec kinases must be regulated by a different set of intramolecular interactions to the Src kinases. We have determined the solution structure of the Tec SH3 domain and have investigated interactions with its PRR, which contains two SH3-binding sites. We demonstrate that in vitro, the Tec PRR can bind in an intramolecular fashion to the SH3. However, the affinity is lower than that for dimerization via reciprocal PRR-SH3 association. Using site-directed mutagenesis we show that both sites can bind the Tec SH3 domain; site 1 (155KTLPPAP161) binds intramolecularly, while site 2 (165KRRPPPPIPP174) cannot and binds in an intermolecular fashion. These distinct roles for the SH3 binding sites in Tec family kinases could be important for protein targeting and enzyme activation.},\\n bibtype = {article},\\n author = {Pursglove, S.E. and Mulhern, T.D. and Mackay, J.P. and Hinds, M.G. and Booker, G.W.},\\n doi = {10.1074/jbc.M108318200},\\n journal = {Journal of Biological Chemistry},\\n number = {1}\\n}\",\"author_short\":[\"Pursglove,  S.\",\"Mulhern,  T.\",\"Mackay,  J.\",\"Hinds,  M.\",\"Booker,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"pursglove-mulhern-mackay-hinds-booker-thesolutionstructureandintramolecularassociationsoftheteckinasesrchomology3domain-2002\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The hydrophobin EAS is largely unstructured in solution and functions by forming amyloid-like structures\",\"type\":\"article\",\"year\":\"2001\",\"pages\":\"83-91\",\"volume\":\"9\",\"id\":\"f7eae917-5560-305a-8f43-505a2f44c290\",\"created\":\"2023-01-10T01:46:32.312Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:32.312Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.\",\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Matthews, J.M. and Winefield, R.D. and Mackay, L.G. and Haverkamp, R.G. and Templeton, M.D.\",\"doi\":\"10.1016/S0969-2126(00)00559-1\",\"journal\":\"Structure\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {The hydrophobin EAS is largely unstructured in solution and functions by forming amyloid-like structures},\\n type = {article},\\n year = {2001},\\n pages = {83-91},\\n volume = {9},\\n id = {f7eae917-5560-305a-8f43-505a2f44c290},\\n created = {2023-01-10T01:46:32.312Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:32.312Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.},\\n bibtype = {article},\\n author = {Mackay, J.P. and Matthews, J.M. and Winefield, R.D. and Mackay, L.G. and Haverkamp, R.G. and Templeton, M.D.},\\n doi = {10.1016/S0969-2126(00)00559-1},\\n journal = {Structure},\\n number = {2}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Matthews,  J.\",\"Winefield,  R.\",\"Mackay,  L.\",\"Haverkamp,  R.\",\"Templeton,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Discovery and Structure of a Potent and Highly Specific Blocker of Insect Calcium Channels\",\"type\":\"article\",\"year\":\"2001\",\"pages\":\"40306-40312\",\"volume\":\"276\",\"id\":\"6c391a59-29b9-321f-858c-b5248f7b56df\",\"created\":\"2023-01-10T01:46:33.265Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:33.265Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"We have isolated a novel family of insect-selective neurotoxins that appear to be the most potent blockers of insect voltage-gated calcium channels reported to date. These toxins display exceptional phylogenetic specificity, with at least a 10,000-fold preference for insect versus vertebrate calcium channels. The structure of one of the toxins reveals a highly structured, disulfide-rich core and a structurally disordered C-terminal extension that is essential for channel blocking activity. Weak structural/functional homology with ω-agatoxin-IVA/B, the prototypic inhibitor of vertebrate P-type calcium channels, suggests that these two toxin families might share a similar mechanism of action despite their vastly different phylogenetic specificities.\",\"bibtype\":\"article\",\"author\":\"Wang, X.-H. and Connor, M. and Wilson, D. and Wilson, H.I. and Nicholson, G.M. and Smith, R. and Shaw, D. and Mackay, J.P. and Alewood, P.F. and Christie, M.J. and Christie, M.J. and King, G.F.\",\"doi\":\"10.1074/jbc.M105206200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"43\",\"bibtex\":\"@article{\\n title = {Discovery and Structure of a Potent and Highly Specific Blocker of Insect Calcium Channels},\\n type = {article},\\n year = {2001},\\n pages = {40306-40312},\\n volume = {276},\\n id = {6c391a59-29b9-321f-858c-b5248f7b56df},\\n created = {2023-01-10T01:46:33.265Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:33.265Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {We have isolated a novel family of insect-selective neurotoxins that appear to be the most potent blockers of insect voltage-gated calcium channels reported to date. These toxins display exceptional phylogenetic specificity, with at least a 10,000-fold preference for insect versus vertebrate calcium channels. The structure of one of the toxins reveals a highly structured, disulfide-rich core and a structurally disordered C-terminal extension that is essential for channel blocking activity. Weak structural/functional homology with ω-agatoxin-IVA/B, the prototypic inhibitor of vertebrate P-type calcium channels, suggests that these two toxin families might share a similar mechanism of action despite their vastly different phylogenetic specificities.},\\n bibtype = {article},\\n author = {Wang, X.-H. and Connor, M. and Wilson, D. and Wilson, H.I. and Nicholson, G.M. and Smith, R. and Shaw, D. and Mackay, J.P. and Alewood, P.F. and Christie, M.J. and Christie, M.J. and King, G.F.},\\n doi = {10.1074/jbc.M105206200},\\n journal = {Journal of Biological Chemistry},\\n number = {43}\\n}\",\"author_short\":[\"Wang,  X.\",\"Connor,  M.\",\"Wilson,  D.\",\"Wilson,  H.\",\"Nicholson,  G.\",\"Smith,  R.\",\"Shaw,  D.\",\"Mackay,  J.\",\"Alewood,  P.\",\"Christie,  M.\",\"Christie,  M.\",\"King,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"wang-connor-wilson-wilson-nicholson-smith-shaw-mackay-etal-discoveryandstructureofapotentandhighlyspecificblockerofinsectcalciumchannels-2001\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Design, production and characterization of FLIN2 and FLIN4: The engineering of intramolecular ldb1:LMO complexes\",\"type\":\"article\",\"year\":\"2001\",\"pages\":\"493-499\",\"volume\":\"14\",\"id\":\"a9b6f2ec-a058-327e-abad-6aa474786e8a\",\"created\":\"2023-01-10T01:46:34.180Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:34.180Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The nuclear LIM-only (LMO) transcription factors LMO2 and LMO4 play important roles in both normal and leukemic T-cell development. LIM domains are cysteine/histidine-rich domains that contain two structural zinc ions and that function as protein-protein adaptors; members of the LMO family each contain two closely spaced LIM domains. These LMO proteins all bind with high affinity to the nuclear protein LIM domain binding protein 1 (ldb1). The LMO-ldb1 interaction is mediated through the N-terminal LIM domain (LIM1) of LMO proteins and a 38-residue region towards the C-terminus of ldb1 [ldb1(LID)]. Unfortunately, recombinant forms of LMO2 and LMO4 have limited solubility and stability, effectively preventing structural analysis. Therefore, we have designed and constructed a fusion protein in which ldb1(LID) and LIM1 of LMO2 can form an intramolecular complex. The engineered protein, FLIN2 (fusion of the LIM interacting domain of ldb1 and the N-terminal LIM domain of LMO2) has been expressed and purified in milligram quantities. FLIN2 is monomeric, contains significant levels of secondary structure and yields a sharp and well-dispersed one-dimensional 1H NMR spectrum. The analogous LMO4 protein, FLIN4, has almost identical properties. These data suggest that we will be able to obtain high-resolution structural information about the LMO-ldb1 interactions.\",\"bibtype\":\"article\",\"author\":\"Deane, J.E. and Sum, E. and Mackay, J.P. and Lindeman, G.J. and Visvader, J.E. and Matthews, J.M.\",\"doi\":\"10.1093/protein/14.7.493\",\"journal\":\"Protein Engineering\",\"number\":\"7\",\"bibtex\":\"@article{\\n title = {Design, production and characterization of FLIN2 and FLIN4: The engineering of intramolecular ldb1:LMO complexes},\\n type = {article},\\n year = {2001},\\n pages = {493-499},\\n volume = {14},\\n id = {a9b6f2ec-a058-327e-abad-6aa474786e8a},\\n created = {2023-01-10T01:46:34.180Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:34.180Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The nuclear LIM-only (LMO) transcription factors LMO2 and LMO4 play important roles in both normal and leukemic T-cell development. LIM domains are cysteine/histidine-rich domains that contain two structural zinc ions and that function as protein-protein adaptors; members of the LMO family each contain two closely spaced LIM domains. These LMO proteins all bind with high affinity to the nuclear protein LIM domain binding protein 1 (ldb1). The LMO-ldb1 interaction is mediated through the N-terminal LIM domain (LIM1) of LMO proteins and a 38-residue region towards the C-terminus of ldb1 [ldb1(LID)]. Unfortunately, recombinant forms of LMO2 and LMO4 have limited solubility and stability, effectively preventing structural analysis. Therefore, we have designed and constructed a fusion protein in which ldb1(LID) and LIM1 of LMO2 can form an intramolecular complex. The engineered protein, FLIN2 (fusion of the LIM interacting domain of ldb1 and the N-terminal LIM domain of LMO2) has been expressed and purified in milligram quantities. FLIN2 is monomeric, contains significant levels of secondary structure and yields a sharp and well-dispersed one-dimensional 1H NMR spectrum. The analogous LMO4 protein, FLIN4, has almost identical properties. These data suggest that we will be able to obtain high-resolution structural information about the LMO-ldb1 interactions.},\\n bibtype = {article},\\n author = {Deane, J.E. and Sum, E. and Mackay, J.P. and Lindeman, G.J. and Visvader, J.E. and Matthews, J.M.},\\n doi = {10.1093/protein/14.7.493},\\n journal = {Protein Engineering},\\n number = {7}\\n}\",\"author_short\":[\"Deane,  J.\",\"Sum,  E.\",\"Mackay,  J.\",\"Lindeman,  G.\",\"Visvader,  J.\",\"Matthews,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"deane-sum-mackay-lindeman-visvader-matthews-designproductionandcharacterizationofflin2andflin4theengineeringofintramolecularldb1lmocomplexes-2001\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of FLIN2, an intramolecular LMO2:ldb1 complex [2]\",\"type\":\"article\",\"year\":\"2001\",\"pages\":\"385-386\",\"volume\":\"21\",\"id\":\"4480e900-209e-3853-9615-5c2f38cb7107\",\"created\":\"2023-01-10T01:46:35.100Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:35.100Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Matthews, J.M. and Visvader, J.E. and Mackay, J.P.\",\"doi\":\"10.1023/A:1013373203772\",\"journal\":\"Journal of Biomolecular NMR\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of FLIN2, an intramolecular LMO2:ldb1 complex [2]},\\n type = {article},\\n year = {2001},\\n pages = {385-386},\\n volume = {21},\\n id = {4480e900-209e-3853-9615-5c2f38cb7107},\\n created = {2023-01-10T01:46:35.100Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:35.100Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Matthews, J.M. and Visvader, J.E. and Mackay, J.P.},\\n doi = {10.1023/A:1013373203772},\\n journal = {Journal of Biomolecular NMR},\\n number = {4}\\n}\",\"author_short\":[\"Matthews,  J.\",\"Visvader,  J.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"matthews-visvader-mackay-sup1suphsup15supnandsup13supcassignmentsofflin2anintramolecularlmo2ldb1complex2-2001\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Probing site specificity of DNA binding metallointercalators by NMR spectroscopy and molecular modeling\",\"type\":\"article\",\"year\":\"2001\",\"pages\":\"4867-4878\",\"volume\":\"40\",\"id\":\"ecd0710f-2f62-38e1-9576-54331818c01f\",\"created\":\"2023-01-10T01:46:35.999Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:35.999Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The molecular recognition of oligonucleotides by chiral ruthenium complexes has been probed by NMR spectroscopy using the template Δ-cis-α- and Δ-cis-β-[Ru(RR-picchxnMe2) (bidentate)]2+, where the bidentate ligand is one of phen (1,10-phenanthroline), dpq (dipyrido[3,2-f:2′,3′-h]quinoxaline), or phi (9,10-phenanthrenequinone diimine) and picchxnMe2 is N,N′-dimethyl-N,N′-di(2-picolyl)-1,2-diaminocyclohexane. By varying only the bidentate ligand in a series of complexes, it was shown that the bidentate alone can alter binding modes. DNA binding studies of the Δ-cis-α,-[Ru(RR-picchxnMe2)(phen)]2+ complex indicate fast exchange kinetics on the chemical shift time scale and a \\\"partial intercalation\\\" mode of binding. This complex binds to [d(CGCGATCGCG)]2 and [d(ATATCGATAT)]2 at AT, TA, and GA sites from the minor groove, as well as to the ends of the oligonucleotide at low temperature. Studies of the Δ-cis-β-[Ru(RR-picchxnMe2)(phen)]2+ complex with [d(CGCGATCGCG)]2 showed that the complex binds only weakly to the ends of the oligonucleotide. The interaction of Δ-cis-α-[Ru(RR-picchxnMe2)-(dpq)]2+ with [d(CGCGATCGCG)]2 showed intermediate exchange kinetics and evidence of minor groove intercalation at the GA base step. In contrast to the phen and dpq complexes, Δ-cis-α- and Δ-cis-β-[Ru-(RR-picchxnMe2)(phi)]2+ showed evidence of major groove binding independent of the metal ion configuration. DNA stabilization induced by complex binding to [d(CGCGATCGCG)]2 (measured as ΔTm) increases in the order phen < dpq and DNA affinity in the order phen < dpq < phi. The groove binding preferences exhibited by the different bidentate ligands is explained with the aid of molecular modeling experiments.\",\"bibtype\":\"article\",\"author\":\"Proudfoot, E.M. and Mackay, J.P. and Karuso, P.\",\"doi\":\"10.1021/bi001655f\",\"journal\":\"Biochemistry\",\"number\":\"15\",\"bibtex\":\"@article{\\n title = {Probing site specificity of DNA binding metallointercalators by NMR spectroscopy and molecular modeling},\\n type = {article},\\n year = {2001},\\n pages = {4867-4878},\\n volume = {40},\\n id = {ecd0710f-2f62-38e1-9576-54331818c01f},\\n created = {2023-01-10T01:46:35.999Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:35.999Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The molecular recognition of oligonucleotides by chiral ruthenium complexes has been probed by NMR spectroscopy using the template Δ-cis-α- and Δ-cis-β-[Ru(RR-picchxnMe2) (bidentate)]2+, where the bidentate ligand is one of phen (1,10-phenanthroline), dpq (dipyrido[3,2-f:2′,3′-h]quinoxaline), or phi (9,10-phenanthrenequinone diimine) and picchxnMe2 is N,N′-dimethyl-N,N′-di(2-picolyl)-1,2-diaminocyclohexane. By varying only the bidentate ligand in a series of complexes, it was shown that the bidentate alone can alter binding modes. DNA binding studies of the Δ-cis-α,-[Ru(RR-picchxnMe2)(phen)]2+ complex indicate fast exchange kinetics on the chemical shift time scale and a \\\"partial intercalation\\\" mode of binding. This complex binds to [d(CGCGATCGCG)]2 and [d(ATATCGATAT)]2 at AT, TA, and GA sites from the minor groove, as well as to the ends of the oligonucleotide at low temperature. Studies of the Δ-cis-β-[Ru(RR-picchxnMe2)(phen)]2+ complex with [d(CGCGATCGCG)]2 showed that the complex binds only weakly to the ends of the oligonucleotide. The interaction of Δ-cis-α-[Ru(RR-picchxnMe2)-(dpq)]2+ with [d(CGCGATCGCG)]2 showed intermediate exchange kinetics and evidence of minor groove intercalation at the GA base step. In contrast to the phen and dpq complexes, Δ-cis-α- and Δ-cis-β-[Ru-(RR-picchxnMe2)(phi)]2+ showed evidence of major groove binding independent of the metal ion configuration. DNA stabilization induced by complex binding to [d(CGCGATCGCG)]2 (measured as ΔTm) increases in the order phen < dpq and DNA affinity in the order phen < dpq < phi. The groove binding preferences exhibited by the different bidentate ligands is explained with the aid of molecular modeling experiments.},\\n bibtype = {article},\\n author = {Proudfoot, E.M. and Mackay, J.P. and Karuso, P.},\\n doi = {10.1021/bi001655f},\\n journal = {Biochemistry},\\n number = {15}\\n}\",\"author_short\":[\"Proudfoot,  E.\",\"Mackay,  J.\",\"Karuso,  P.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"proudfoot-mackay-karuso-probingsitespecificityofdnabindingmetallointercalatorsbynmrspectroscopyandmolecularmodeling-2001\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The N-terminal Zinc Finger of the Erythroid Transcription Factor GATA-1 Binds GATC Motifs in DNA\",\"type\":\"article\",\"year\":\"2001\",\"pages\":\"35794-35801\",\"volume\":\"276\",\"id\":\"668a6be7-8063-3f78-a869-df88d49978b0\",\"created\":\"2023-01-10T01:46:36.902Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:36.902Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The mammalian transcription factor GATA-1 is required for normal erythroid and megakaryocytic development. GATA-1 contains two zinc fingers, the C-terminal finger, which is known to bind (A/T)GATA(A/G) motifs in DNA and the N-finger, which is important for interacting with co-regulatory proteins such as Friend of GATA (FOG). We now show that, like the C-finger, the N-finger of GATA-1 is also capable of binding DNA but recognizes distinct sequences with the core GATC. We demonstrate that the GATA-1 N-finger can bind these sequences in vitro and that in cellular assays, GATA-1 can activate promoters containing GATC motifs. Experiments with mutant GATA-1 proteins confirm the importance of the N-finger, as the C-finger is not required for transactivation from GATC sites. Recently four naturally occurring mutations in GATA-1 have been shown to be associated with familial blood disorders. These mutations all map to the N-finger domain. We have investigated the effect of these mutations on the recognition of GATC sites by the N-finger and show that one mutation R216Q abolishes DNA binding, whereas the others have only minor effects.\",\"bibtype\":\"article\",\"author\":\"Newton, A. and Mackay, J. and Crossley, M.\",\"doi\":\"10.1074/jbc.M106256200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"38\",\"bibtex\":\"@article{\\n title = {The N-terminal Zinc Finger of the Erythroid Transcription Factor GATA-1 Binds GATC Motifs in DNA},\\n type = {article},\\n year = {2001},\\n pages = {35794-35801},\\n volume = {276},\\n id = {668a6be7-8063-3f78-a869-df88d49978b0},\\n created = {2023-01-10T01:46:36.902Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:36.902Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The mammalian transcription factor GATA-1 is required for normal erythroid and megakaryocytic development. GATA-1 contains two zinc fingers, the C-terminal finger, which is known to bind (A/T)GATA(A/G) motifs in DNA and the N-finger, which is important for interacting with co-regulatory proteins such as Friend of GATA (FOG). We now show that, like the C-finger, the N-finger of GATA-1 is also capable of binding DNA but recognizes distinct sequences with the core GATC. We demonstrate that the GATA-1 N-finger can bind these sequences in vitro and that in cellular assays, GATA-1 can activate promoters containing GATC motifs. Experiments with mutant GATA-1 proteins confirm the importance of the N-finger, as the C-finger is not required for transactivation from GATC sites. Recently four naturally occurring mutations in GATA-1 have been shown to be associated with familial blood disorders. These mutations all map to the N-finger domain. We have investigated the effect of these mutations on the recognition of GATC sites by the N-finger and show that one mutation R216Q abolishes DNA binding, whereas the others have only minor effects.},\\n bibtype = {article},\\n author = {Newton, A. and Mackay, J. and Crossley, M.},\\n doi = {10.1074/jbc.M106256200},\\n journal = {Journal of Biological Chemistry},\\n number = {38}\\n}\",\"author_short\":[\"Newton,  A.\",\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"newton-mackay-crossley-thenterminalzincfingeroftheerythroidtranscriptionfactorgata1bindsgatcmotifsindna-2001\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The transactivation domain within cysteine/histidine-rich region 1 of CBP comprises two novel zinc-binding modules\",\"type\":\"article\",\"year\":\"2000\",\"pages\":\"15128-15134\",\"volume\":\"275\",\"id\":\"912798e3-760b-3e6e-bdd0-d9324be5e544\",\"created\":\"2023-01-10T01:46:37.833Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:37.833Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"cAMP-response element-binding protein-binding protein (CBP) is a transcriptional coactivator that interacts with a number of DNA-binding proteins and cofactor proteins involved in the regulation of transcription. Relatively little is known about the structure of CBP, but it has been noted that it contains three domains that are rich in cysteine and histidine (CH1, CH2, and CH3). The sequence of CH2 conforms to that of a leukemia-associated protein domain (PHD finger), and it has been postulated that this and both CH1 and CH3 may be zinc finger domains. This has not, however, been demonstrated experimentally. We have studied CH1 and show that it is composed of two novel zinc-binding modules, which we term 'zinc bundles.' Each bundle contains the sequence Cys-X4-Cys-X8-His-X3-Cys, and we show that a synthetic peptide comprising one zinc bundle from CH1 can fold in a zinc- dependent manner. CH3 also appears to contain two zinc bundles, one with the variant sequence Cys-X2-Cys-X9-His-X3-Cys, and we demonstrate that this variant motif also undergoes Zn(II)-induced folding. CH1 acts as a transcriptional activation domain in cellular assays. We show that mutations in any of the four zinc-chelating residues in either zinc bundle of CH1 significantly impair this activity and that these mutations also interfere with certain protein-protein interactions mediated by CH1. Our results indicate that CBP is a genuine zinc-binding protein and introduce zinc bundles as novel protein interaction domains.\",\"bibtype\":\"article\",\"author\":\"Newton, A.L. and Sharped, B.K. and Kwan, A. and Mackay, J.P. and Crossley, M.\",\"doi\":\"10.1074/jbc.M910396199\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"20\",\"bibtex\":\"@article{\\n title = {The transactivation domain within cysteine/histidine-rich region 1 of CBP comprises two novel zinc-binding modules},\\n type = {article},\\n year = {2000},\\n pages = {15128-15134},\\n volume = {275},\\n id = {912798e3-760b-3e6e-bdd0-d9324be5e544},\\n created = {2023-01-10T01:46:37.833Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:37.833Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {cAMP-response element-binding protein-binding protein (CBP) is a transcriptional coactivator that interacts with a number of DNA-binding proteins and cofactor proteins involved in the regulation of transcription. Relatively little is known about the structure of CBP, but it has been noted that it contains three domains that are rich in cysteine and histidine (CH1, CH2, and CH3). The sequence of CH2 conforms to that of a leukemia-associated protein domain (PHD finger), and it has been postulated that this and both CH1 and CH3 may be zinc finger domains. This has not, however, been demonstrated experimentally. We have studied CH1 and show that it is composed of two novel zinc-binding modules, which we term 'zinc bundles.' Each bundle contains the sequence Cys-X4-Cys-X8-His-X3-Cys, and we show that a synthetic peptide comprising one zinc bundle from CH1 can fold in a zinc- dependent manner. CH3 also appears to contain two zinc bundles, one with the variant sequence Cys-X2-Cys-X9-His-X3-Cys, and we demonstrate that this variant motif also undergoes Zn(II)-induced folding. CH1 acts as a transcriptional activation domain in cellular assays. We show that mutations in any of the four zinc-chelating residues in either zinc bundle of CH1 significantly impair this activity and that these mutations also interfere with certain protein-protein interactions mediated by CH1. Our results indicate that CBP is a genuine zinc-binding protein and introduce zinc bundles as novel protein interaction domains.},\\n bibtype = {article},\\n author = {Newton, A.L. and Sharped, B.K. and Kwan, A. and Mackay, J.P. and Crossley, M.},\\n doi = {10.1074/jbc.M910396199},\\n journal = {Journal of Biological Chemistry},\\n number = {20}\\n}\",\"author_short\":[\"Newton,  A.\",\"Sharped,  B.\",\"Kwan,  A.\",\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"newton-sharped-kwan-mackay-crossley-thetransactivationdomainwithincysteinehistidinerichregion1ofcbpcomprisestwonovelzincbindingmodules-2000\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The core of the respiratory syncytial virus fusion protein is a trimeric coiled coil\",\"type\":\"article\",\"year\":\"2000\",\"pages\":\"5911-5920\",\"volume\":\"74\",\"id\":\"82505942-a661-3722-bf77-e61b0007b83b\",\"created\":\"2023-01-10T01:46:38.745Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:38.745Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Entry into the host cell by enveloped viruses is mediated by fusion (F) or transmembrane glycoproteins. Many of these proteins share a fold comprising a trimer of antiparallel coiled-coil heterodimers, where the heterodimers are formed by two discontinuous heptad repeat motifs within the proteolytically processed chain. The F protein of human respiratory syncytial virus (RSV; the major cause of lower respiratory tract infections in infants) contains two corresponding regions that are predicted to form coiled coils (HR1 and HR2), together with a third predicted heptad repeat (HR3) located in a nonhomologous position. In order to probe the structures of these three domains and ascertain the nature of the interactions between them, we have studied the isolated HR1, HR2, and HR3 domains of RSV F by using a range of biophysical techniques, including circular dichroism, nuclear magnetic resonance spectroscopy, and sedimentation equilibrium. HR1 forms a symmetrical, trimeric coiled coil in solution (K3 ≃ 2.2 x 1011 M-2) which interacts with HR2 to form a 3:3 hexamer. The HR1-HR2 interaction domains have been mapped using limited proteolysis, reversed-phase high- performance liquid chromatography, and electrospray-mass spectrometry. HR2 in isolation exists as a largely unstructured monomer, although it exhibits a tendency to form aggregates with β-sheet-like characteristics. Only a small increase in α-helical content was observed upon the formation of the hexamer. This suggests that the RSV F glycoprotein contains a domain that closely resembles the core structure of the simian parainfluenza virus 5 fusion protein (K. A. Baker, R. E. Dutch, R. A. Lamb, and T. S. Jardetzky, Mol. Cell 3:309-319, 1999). Finally, HR3 forms weak α-helical homodimers that do not appear to interact with HR1, HR2, or the HR1-HR2 complex. The results of these studies support the idea that viral fusion proteins have a common core architecture.\",\"bibtype\":\"article\",\"author\":\"Matthews, J.M. and Young, T.F. and Tucker, S.P. and Mackay, J.P.\",\"doi\":\"10.1128/JVI.74.13.5911-5920.2000\",\"journal\":\"Journal of Virology\",\"number\":\"13\",\"bibtex\":\"@article{\\n title = {The core of the respiratory syncytial virus fusion protein is a trimeric coiled coil},\\n type = {article},\\n year = {2000},\\n pages = {5911-5920},\\n volume = {74},\\n id = {82505942-a661-3722-bf77-e61b0007b83b},\\n created = {2023-01-10T01:46:38.745Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:38.745Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Entry into the host cell by enveloped viruses is mediated by fusion (F) or transmembrane glycoproteins. Many of these proteins share a fold comprising a trimer of antiparallel coiled-coil heterodimers, where the heterodimers are formed by two discontinuous heptad repeat motifs within the proteolytically processed chain. The F protein of human respiratory syncytial virus (RSV; the major cause of lower respiratory tract infections in infants) contains two corresponding regions that are predicted to form coiled coils (HR1 and HR2), together with a third predicted heptad repeat (HR3) located in a nonhomologous position. In order to probe the structures of these three domains and ascertain the nature of the interactions between them, we have studied the isolated HR1, HR2, and HR3 domains of RSV F by using a range of biophysical techniques, including circular dichroism, nuclear magnetic resonance spectroscopy, and sedimentation equilibrium. HR1 forms a symmetrical, trimeric coiled coil in solution (K3 ≃ 2.2 x 1011 M-2) which interacts with HR2 to form a 3:3 hexamer. The HR1-HR2 interaction domains have been mapped using limited proteolysis, reversed-phase high- performance liquid chromatography, and electrospray-mass spectrometry. HR2 in isolation exists as a largely unstructured monomer, although it exhibits a tendency to form aggregates with β-sheet-like characteristics. Only a small increase in α-helical content was observed upon the formation of the hexamer. This suggests that the RSV F glycoprotein contains a domain that closely resembles the core structure of the simian parainfluenza virus 5 fusion protein (K. A. Baker, R. E. Dutch, R. A. Lamb, and T. S. Jardetzky, Mol. Cell 3:309-319, 1999). Finally, HR3 forms weak α-helical homodimers that do not appear to interact with HR1, HR2, or the HR1-HR2 complex. The results of these studies support the idea that viral fusion proteins have a common core architecture.},\\n bibtype = {article},\\n author = {Matthews, J.M. and Young, T.F. and Tucker, S.P. and Mackay, J.P.},\\n doi = {10.1128/JVI.74.13.5911-5920.2000},\\n journal = {Journal of Virology},\\n number = {13}\\n}\",\"author_short\":[\"Matthews,  J.\",\"Young,  T.\",\"Tucker,  S.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"matthews-young-tucker-mackay-thecoreoftherespiratorysyncytialvirusfusionproteinisatrimericcoiledcoil-2000\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Solution structures of two CCHC zinc fingers from the FOG family protein U-shaped that mediate protein-protein interactions\",\"type\":\"article\",\"year\":\"2000\",\"pages\":\"1157-1166\",\"volume\":\"8\",\"id\":\"4220c92e-ca96-3078-a476-5f478fe44cdb\",\"created\":\"2023-01-10T01:46:39.669Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:39.669Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.\",\"bibtype\":\"article\",\"author\":\"Chu Kong Liew, undefined and Kowalski, K. and Fox, A.H. and Newton, A. and Sharpe, B.K. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1016/S0969-2126(00)00527-X\",\"journal\":\"Structure\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Solution structures of two CCHC zinc fingers from the FOG family protein U-shaped that mediate protein-protein interactions},\\n type = {article},\\n year = {2000},\\n pages = {1157-1166},\\n volume = {8},\\n id = {4220c92e-ca96-3078-a476-5f478fe44cdb},\\n created = {2023-01-10T01:46:39.669Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:39.669Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.},\\n bibtype = {article},\\n author = {Chu Kong Liew, undefined and Kowalski, K. and Fox, A.H. and Newton, A. and Sharpe, B.K. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1016/S0969-2126(00)00527-X},\\n journal = {Structure},\\n number = {11}\\n}\",\"author_short\":[\"Chu Kong Liew\",\"Kowalski,  K.\",\"Fox,  A.\",\"Newton,  A.\",\"Sharpe,  B.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"chukongliew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Interfacial asparagine residues within an amide tetrad contribute to Max helix-loop-helix leucine zipper homodimer stability\",\"type\":\"article\",\"year\":\"2000\",\"pages\":\"37454-37461\",\"volume\":\"275\",\"id\":\"44466ab6-2aa6-3104-b086-fd106c65ff76\",\"created\":\"2023-01-10T01:46:40.592Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:40.592Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The transcription factor Max is the obligate dimerization partner of the Myc oncoprotein. The pivotal role of Max within the Myc regulatory network is dependent upon its ability to dimerize via the helix-loop-helix leucine zipper domain. The Max homodimer contains a tetrad of polar residues at the interface of the leucine zipper domain. A conserved interfacial Asn residue at an equivalent position in two other leucine zipper proteins has been shown to decrease homodimer stability. The unusual arrangement of this Gln-Asn/Gln'-Asn' tetrad prompted us to investigate whether Asn92 plays a similar role in destabilizing the Max homodimer. This residue was sequentially replaced with aliphatic and charged residues. Thermal denaturation, redox time course and analytical ultracentrifugation studies show that the N92V mutation does not increase homodimer stability. Replacing this residue with negatively charged side chains in N92D and N92E destabilizes the mutant homodimer. Further replacement of Gln91 indicated that H bonding between Gln91 and Asn92 residues is not significant to the stability of the native protein. These data collectively demonstrate the central role of Asn92 in homodimer interactions. Molecular modelling studies illustrate the favorable packing of the native Asn residue at the dimer interface compared with that of the mutant Max peptides.\",\"bibtype\":\"article\",\"author\":\"Tchan, M.C. and Choy, K.J. and Mackay, J.P. and Lyons, A.T.L. and Bains, N.P.S. and Weiss, A.S.\",\"doi\":\"10.1074/jbc.M004264200\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"48\",\"bibtex\":\"@article{\\n title = {Interfacial asparagine residues within an amide tetrad contribute to Max helix-loop-helix leucine zipper homodimer stability},\\n type = {article},\\n year = {2000},\\n pages = {37454-37461},\\n volume = {275},\\n id = {44466ab6-2aa6-3104-b086-fd106c65ff76},\\n created = {2023-01-10T01:46:40.592Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:40.592Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The transcription factor Max is the obligate dimerization partner of the Myc oncoprotein. The pivotal role of Max within the Myc regulatory network is dependent upon its ability to dimerize via the helix-loop-helix leucine zipper domain. The Max homodimer contains a tetrad of polar residues at the interface of the leucine zipper domain. A conserved interfacial Asn residue at an equivalent position in two other leucine zipper proteins has been shown to decrease homodimer stability. The unusual arrangement of this Gln-Asn/Gln'-Asn' tetrad prompted us to investigate whether Asn92 plays a similar role in destabilizing the Max homodimer. This residue was sequentially replaced with aliphatic and charged residues. Thermal denaturation, redox time course and analytical ultracentrifugation studies show that the N92V mutation does not increase homodimer stability. Replacing this residue with negatively charged side chains in N92D and N92E destabilizes the mutant homodimer. Further replacement of Gln91 indicated that H bonding between Gln91 and Asn92 residues is not significant to the stability of the native protein. These data collectively demonstrate the central role of Asn92 in homodimer interactions. Molecular modelling studies illustrate the favorable packing of the native Asn residue at the dimer interface compared with that of the mutant Max peptides.},\\n bibtype = {article},\\n author = {Tchan, M.C. and Choy, K.J. and Mackay, J.P. and Lyons, A.T.L. and Bains, N.P.S. and Weiss, A.S.},\\n doi = {10.1074/jbc.M004264200},\\n journal = {Journal of Biological Chemistry},\\n number = {48}\\n}\",\"author_short\":[\"Tchan,  M.\",\"Choy,  K.\",\"Mackay,  J.\",\"Lyons,  A.\",\"Bains,  N.\",\"Weiss,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"tchan-choy-mackay-lyons-bains-weiss-interfacialasparagineresidueswithinanamidetetradcontributetomaxhelixloophelixleucinezipperhomodimerstability-2000\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"A class of zinc fingers involved in protein-protein interactions\",\"type\":\"article\",\"year\":\"2000\",\"pages\":\"1030-1038\",\"volume\":\"267\",\"id\":\"3aade15c-920e-3fd6-9d2d-2812368f8f88\",\"created\":\"2023-01-10T01:46:41.497Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:41.497Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Zinc fingers (ZnFs) are extremely common protein domains. Several classes of ZnFs are distinguished by the nature and spacing of their zinc- coordinating residues. While the structure and function of some ZnFs are well characterized, many others have been identified only through their amino acid sequence. A number of proteins contain a conserved C-X2-C-X12-H-X1-5-C sequence, which is similar to the spacing observed for the 'classic' CCHH ZnFs. Although these domains have been implicated in protein-protein (and not protein-nucleic acid) interactions, nothing is known about their structure or function at a molecular level. Here, we address this problem through the expression and biophysical characterization of several CCHC-type zinc fingers from the erythroid transcription factor FOG and the related Drosophila protein U-shaped. Each of these domains does indeed fold in a zinc-dependent fashion, coordinating the metal in a tetrahedral manner through the sidechains of one histidine and three cysteine residues, and forming extremely thermostable structures. Analysis of CD spectra suggests an overall fold similar to that of the CCHH fingers, and indeed a point mutant of FOG-F1 in which the final cysteine residue is replaced by histidine remains capable of folding. However, the CCHC (as opposed to CCHH) motif is a prerequisite for GATA-1 binding activity, demonstrating that CCHC and CCHH topologies are not interchangeable. This demonstration that members of a structurally distinct subclass of genuine zinc finger domains are involved in the mediation of protein-protein interactions has implications for the prediction of protein function from nucleotide sequences.\",\"bibtype\":\"article\",\"author\":\"Matthews, J.M. and Kowalski, K. and Liew, C.K. and Sharpe, B.K. and Fox, A.H. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1046/j.1432-1327.2000.01095.x\",\"journal\":\"European Journal of Biochemistry\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {A class of zinc fingers involved in protein-protein interactions},\\n type = {article},\\n year = {2000},\\n pages = {1030-1038},\\n volume = {267},\\n id = {3aade15c-920e-3fd6-9d2d-2812368f8f88},\\n created = {2023-01-10T01:46:41.497Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:41.497Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Zinc fingers (ZnFs) are extremely common protein domains. Several classes of ZnFs are distinguished by the nature and spacing of their zinc- coordinating residues. While the structure and function of some ZnFs are well characterized, many others have been identified only through their amino acid sequence. A number of proteins contain a conserved C-X2-C-X12-H-X1-5-C sequence, which is similar to the spacing observed for the 'classic' CCHH ZnFs. Although these domains have been implicated in protein-protein (and not protein-nucleic acid) interactions, nothing is known about their structure or function at a molecular level. Here, we address this problem through the expression and biophysical characterization of several CCHC-type zinc fingers from the erythroid transcription factor FOG and the related Drosophila protein U-shaped. Each of these domains does indeed fold in a zinc-dependent fashion, coordinating the metal in a tetrahedral manner through the sidechains of one histidine and three cysteine residues, and forming extremely thermostable structures. Analysis of CD spectra suggests an overall fold similar to that of the CCHH fingers, and indeed a point mutant of FOG-F1 in which the final cysteine residue is replaced by histidine remains capable of folding. However, the CCHC (as opposed to CCHH) motif is a prerequisite for GATA-1 binding activity, demonstrating that CCHC and CCHH topologies are not interchangeable. This demonstration that members of a structurally distinct subclass of genuine zinc finger domains are involved in the mediation of protein-protein interactions has implications for the prediction of protein function from nucleotide sequences.},\\n bibtype = {article},\\n author = {Matthews, J.M. and Kowalski, K. and Liew, C.K. and Sharpe, B.K. and Fox, A.H. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1046/j.1432-1327.2000.01095.x},\\n journal = {European Journal of Biochemistry},\\n number = {4}\\n}\",\"author_short\":[\"Matthews,  J.\",\"Kowalski,  K.\",\"Liew,  C.\",\"Sharpe,  B.\",\"Fox,  A.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"matthews-kowalski-liew-sharpe-fox-crossley-mackay-aclassofzincfingersinvolvedinproteinproteininteractions-2000\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG\",\"type\":\"article\",\"year\":\"1999\",\"pages\":\"249-262\",\"volume\":\"13\",\"id\":\"649a742c-0f2a-3975-8e3d-1abdb42ec917\",\"created\":\"2023-01-10T01:46:42.416Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:42.416Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted β-hairpins and a single α-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA- 1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.\",\"bibtype\":\"article\",\"author\":\"Kowalski, K. and Czolij, R. and King, G.F. and Crossley, M. and Mackay, J.P.\",\"doi\":\"10.1023/A:1008309602929\",\"journal\":\"Journal of Biomolecular NMR\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG},\\n type = {article},\\n year = {1999},\\n pages = {249-262},\\n volume = {13},\\n id = {649a742c-0f2a-3975-8e3d-1abdb42ec917},\\n created = {2023-01-10T01:46:42.416Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:42.416Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted β-hairpins and a single α-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA- 1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.},\\n bibtype = {article},\\n author = {Kowalski, K. and Czolij, R. and King, G.F. and Crossley, M. and Mackay, J.P.},\\n doi = {10.1023/A:1008309602929},\\n journal = {Journal of Biomolecular NMR},\\n number = {3}\\n}\",\"author_short\":[\"Kowalski,  K.\",\"Czolij,  R.\",\"King,  G.\",\"Crossley,  M.\",\"Mackay,  J.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"The dimerization and topological specificity functions of MinE reside in a structurally autonomous C-terminal domain\",\"type\":\"article\",\"year\":\"1999\",\"pages\":\"1161-1169\",\"volume\":\"31\",\"id\":\"cd93d5ad-9177-3466-a3fb-e5c4f562db3d\",\"created\":\"2023-01-10T01:46:43.487Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:43.487Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Correct placement of the division septum in Escherichia coli requires the co-ordinated action of three proteins, MinC, MinD and MinE. MinC and MinD interact to form a non-specific division inhibitor that blocks septation at all potential division sites. MinE is able to antagonize MinCD in a topologically sensitive manner, as it restricts MinCD activity to the unwanted division sites at the cell poles. Here, we show that the topological specificity function of MinE residues in a structurally autonomous, trypsin-resistant domain comprising residues 31-88. Nuclear magnetic resonance (NMR) and circular dichroic spectroscopy indicate that this domain includes both α and β secondary structure, while analytical ultracentrifugation reveals that it also contains a region responsible for MinE homodimerization. While trypsin digestion indicates that the anti-MinCD domain of MinE (residues 1-22) does not form a tightly folded structural domain, NMR analysis of a peptide corresponding to MinE1-22 indicates that this region forms a nascent helix in which the peptide rapidly interconverts between disordered (random coil) and α-helical conformations. This suggests that the N-terminal region of MinE may be poised to adopt an α-helical conformation when it interacts with the target of its anti-MinCD activity, presumably MinD.\",\"bibtype\":\"article\",\"author\":\"King, G.F. and Rowland, S.L. and Pan, B. and Mackay, J.P. and Mullen, G.P. and Rothfield, L.I.\",\"doi\":\"10.1046/j.1365-2958.1999.01256.x\",\"journal\":\"Molecular Microbiology\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {The dimerization and topological specificity functions of MinE reside in a structurally autonomous C-terminal domain},\\n type = {article},\\n year = {1999},\\n pages = {1161-1169},\\n volume = {31},\\n id = {cd93d5ad-9177-3466-a3fb-e5c4f562db3d},\\n created = {2023-01-10T01:46:43.487Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:43.487Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Correct placement of the division septum in Escherichia coli requires the co-ordinated action of three proteins, MinC, MinD and MinE. MinC and MinD interact to form a non-specific division inhibitor that blocks septation at all potential division sites. MinE is able to antagonize MinCD in a topologically sensitive manner, as it restricts MinCD activity to the unwanted division sites at the cell poles. Here, we show that the topological specificity function of MinE residues in a structurally autonomous, trypsin-resistant domain comprising residues 31-88. Nuclear magnetic resonance (NMR) and circular dichroic spectroscopy indicate that this domain includes both α and β secondary structure, while analytical ultracentrifugation reveals that it also contains a region responsible for MinE homodimerization. While trypsin digestion indicates that the anti-MinCD domain of MinE (residues 1-22) does not form a tightly folded structural domain, NMR analysis of a peptide corresponding to MinE1-22 indicates that this region forms a nascent helix in which the peptide rapidly interconverts between disordered (random coil) and α-helical conformations. This suggests that the N-terminal region of MinE may be poised to adopt an α-helical conformation when it interacts with the target of its anti-MinCD activity, presumably MinD.},\\n bibtype = {article},\\n author = {King, G.F. and Rowland, S.L. and Pan, B. and Mackay, J.P. and Mullen, G.P. and Rothfield, L.I.},\\n doi = {10.1046/j.1365-2958.1999.01256.x},\\n journal = {Molecular Microbiology},\\n number = {4}\\n}\",\"author_short\":[\"King,  G.\",\"Rowland,  S.\",\"Pan,  B.\",\"Mackay,  J.\",\"Mullen,  G.\",\"Rothfield,  L.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"king-rowland-pan-mackay-mullen-rothfield-thedimerizationandtopologicalspecificityfunctionsofmineresideinastructurallyautonomouscterminaldomain-1999\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Autonomous folding of a peptide corresponding to the N-terminal β- hairpin from ubiquitin\",\"type\":\"article\",\"year\":\"1999\",\"pages\":\"1320-1331\",\"volume\":\"8\",\"id\":\"2fa2cd79-9bc1-3f5c-8753-6943e5016588\",\"created\":\"2023-01-10T01:46:44.399Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:44.399Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The N-terminal 17 residues of ubiquitin have been shown by 1H NMR to fold autonomously into a β-hairpin structure in aqueous solution. This structure has a specific, native-like register, though side-chain contacts differ in detail from those observed in the intact protein. An autonomously folding hairpin has previously been identified in the case of streptococcal protein G, which is structurally homologous with ubiquitin, but remarkably, the two are not in topologically equivalent positions in the fold. This suggests that the organization of folding may be quite different for proteins sharing similar tertiary structures. Two smaller peptides have also been studied, corresponding to the isolated arms of the N-terminal hairpin of ubiquitin, and significant differences from simple random coil predictions observed in the spectra of these subfragments, suggestive of significant limitation of the backbone conformational space sampled, presumably as a consequence of the strongly β-structure favoring composition of the sequences. This illustrates the ability of local sequence elements to express a propensity for β-structure even in the absence of actual sheet formation. Attempts were made to estimate the population of the folded state of the hairpin, in terms of a simple two-state folding model. Using published 'random coil' values to model the unfolded state, and values derived from native ubiquitin for the putative unique, folded state, it was found that the apparent population varied widely for different residues and with different NMR parameters. Use of the spectra of the subfragment peptides to provide a more realistic model of the unfolded state led to better agreement in the estimates that could be obtained from chemical shift and coupling constant measurements, while making it clear that some other approaches to population estimation could not give meaningful results, because of the tendency to populate the β-region of conformational space even in the absence of the hairpin structure.\",\"bibtype\":\"article\",\"author\":\"Zerella, R. and Evans, P.A. and Ionides, J.M.C. and Packman, L.C. and Trotter, B.W. and Mackay, J.P. and Williams, D.H.\",\"doi\":\"10.1110/ps.8.6.1320\",\"journal\":\"Protein Science\",\"number\":\"6\",\"bibtex\":\"@article{\\n title = {Autonomous folding of a peptide corresponding to the N-terminal β- hairpin from ubiquitin},\\n type = {article},\\n year = {1999},\\n pages = {1320-1331},\\n volume = {8},\\n id = {2fa2cd79-9bc1-3f5c-8753-6943e5016588},\\n created = {2023-01-10T01:46:44.399Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:44.399Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The N-terminal 17 residues of ubiquitin have been shown by 1H NMR to fold autonomously into a β-hairpin structure in aqueous solution. This structure has a specific, native-like register, though side-chain contacts differ in detail from those observed in the intact protein. An autonomously folding hairpin has previously been identified in the case of streptococcal protein G, which is structurally homologous with ubiquitin, but remarkably, the two are not in topologically equivalent positions in the fold. This suggests that the organization of folding may be quite different for proteins sharing similar tertiary structures. Two smaller peptides have also been studied, corresponding to the isolated arms of the N-terminal hairpin of ubiquitin, and significant differences from simple random coil predictions observed in the spectra of these subfragments, suggestive of significant limitation of the backbone conformational space sampled, presumably as a consequence of the strongly β-structure favoring composition of the sequences. This illustrates the ability of local sequence elements to express a propensity for β-structure even in the absence of actual sheet formation. Attempts were made to estimate the population of the folded state of the hairpin, in terms of a simple two-state folding model. Using published 'random coil' values to model the unfolded state, and values derived from native ubiquitin for the putative unique, folded state, it was found that the apparent population varied widely for different residues and with different NMR parameters. Use of the spectra of the subfragment peptides to provide a more realistic model of the unfolded state led to better agreement in the estimates that could be obtained from chemical shift and coupling constant measurements, while making it clear that some other approaches to population estimation could not give meaningful results, because of the tendency to populate the β-region of conformational space even in the absence of the hairpin structure.},\\n bibtype = {article},\\n author = {Zerella, R. and Evans, P.A. and Ionides, J.M.C. and Packman, L.C. and Trotter, B.W. and Mackay, J.P. and Williams, D.H.},\\n doi = {10.1110/ps.8.6.1320},\\n journal = {Protein Science},\\n number = {6}\\n}\",\"author_short\":[\"Zerella,  R.\",\"Evans,  P.\",\"Ionides,  J.\",\"Packman,  L.\",\"Trotter,  B.\",\"Mackay,  J.\",\"Williams,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"zerella-evans-ionides-packman-trotter-mackay-williams-autonomousfoldingofapeptidecorrespondingtothenterminalhairpinfromubiquitin-1999\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Transcriptional cofactors of the FOG family interact with GATA proteins by means of multiple zinc fingers\",\"type\":\"article\",\"year\":\"1999\",\"pages\":\"2812-2822\",\"volume\":\"18\",\"id\":\"008d2a64-1e9c-3d79-90e8-1d4d6afcbc71\",\"created\":\"2023-01-10T01:46:45.293Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:45.293Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Friend of GATA-1 (FOG-1) is a zinc finger protein that has been shown to interact physically with the erythroid DNA-binding protein GATA-1 and modulate its transcriptional activity. Recently, two new members of the FOG family have been identified: a mammalian protein, FOG-2, that also associates with GATA-1 and other mammalian GATA factors; and U-shaped, a Drosophila protein that interacts with the Drosophila GATA protein Pannier. FOG proteins contain multiple zinc fingers and it has been shown previously that the sixth finger of FOG-1 interacts specifically with the N-finger but not the C-finger of GATA-1. Here we show that fingers 1, 5 and 9 of FOG-1 also interact with the N-finger of GATA-1 and that FOG-2 and U-shaped also contain multiple GATA-interacting fingers. We define the key contact residues and show that these residues are highly conserved in GATA-interacting fingers. We examine the effect of selectively mutating the four interacting fingers of FOG-1 and show that each contributes to FOG-1's ability to modulate GATA-1 activity. Finally, we show that FOG-1 can repress GATA-1-mediated activation and present evidence that this ability involves the recently described CtBP co-repressor proteins that recognize all known FOG proteins.\",\"bibtype\":\"article\",\"author\":\"Fox, A.H. and Liew, C. and Holmes, M. and Kowalski, K. and Mackay, J. and Crossley, M.\",\"doi\":\"10.1093/emboj/18.10.2812\",\"journal\":\"EMBO Journal\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {Transcriptional cofactors of the FOG family interact with GATA proteins by means of multiple zinc fingers},\\n type = {article},\\n year = {1999},\\n pages = {2812-2822},\\n volume = {18},\\n id = {008d2a64-1e9c-3d79-90e8-1d4d6afcbc71},\\n created = {2023-01-10T01:46:45.293Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:45.293Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Friend of GATA-1 (FOG-1) is a zinc finger protein that has been shown to interact physically with the erythroid DNA-binding protein GATA-1 and modulate its transcriptional activity. Recently, two new members of the FOG family have been identified: a mammalian protein, FOG-2, that also associates with GATA-1 and other mammalian GATA factors; and U-shaped, a Drosophila protein that interacts with the Drosophila GATA protein Pannier. FOG proteins contain multiple zinc fingers and it has been shown previously that the sixth finger of FOG-1 interacts specifically with the N-finger but not the C-finger of GATA-1. Here we show that fingers 1, 5 and 9 of FOG-1 also interact with the N-finger of GATA-1 and that FOG-2 and U-shaped also contain multiple GATA-interacting fingers. We define the key contact residues and show that these residues are highly conserved in GATA-interacting fingers. We examine the effect of selectively mutating the four interacting fingers of FOG-1 and show that each contributes to FOG-1's ability to modulate GATA-1 activity. Finally, we show that FOG-1 can repress GATA-1-mediated activation and present evidence that this ability involves the recently described CtBP co-repressor proteins that recognize all known FOG proteins.},\\n bibtype = {article},\\n author = {Fox, A.H. and Liew, C. and Holmes, M. and Kowalski, K. and Mackay, J. and Crossley, M.},\\n doi = {10.1093/emboj/18.10.2812},\\n journal = {EMBO Journal},\\n number = {10}\\n}\",\"author_short\":[\"Fox,  A.\",\"Liew,  C.\",\"Holmes,  M.\",\"Kowalski,  K.\",\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"fox-liew-holmes-kowalski-mackay-crossley-transcriptionalcofactorsofthefogfamilyinteractwithgataproteinsbymeansofmultiplezincfingers-1999\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Use of altered specificity mutants to probe a specific protein-protein interaction in differentiation: The GATA-1:FOG complex\",\"type\":\"article\",\"year\":\"1999\",\"pages\":\"219-228\",\"volume\":\"3\",\"id\":\"ed2ce0ef-2974-32de-ac4b-b652a7d61a3b\",\"created\":\"2023-01-10T01:46:46.200Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:46.200Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"GATA-1 and FOG (Friend of GATA-1) are each essential for erythroid and megakaryocyte development. FOG, a zinc finger protein, interacts with the amino (N) finger of GATA-1 and cooperates with GATA-1 to promote differentiation. To determine whether this interaction is critical for GATA- 1 action, we selected GATA-1 mutants in yeast that fail to interact with FOG but retain normal DNA binding, as well a compensatory FOG mutant that restores interaction. These novel GATA-1 mutants do not promote erythroid differentiation of GATA-1- erythroid cells. Differentiation is rescued by the second-site FOG mutant. Thus, interaction of FOG with GATA-1 is essential for the function of GATA-1 in erythroid differentiation. These findings provide a paradigm for dissecting protein-protein associations involved in mammalian development.\",\"bibtype\":\"article\",\"author\":\"Crispino, J.D. and Lodish, M.B. and MacKay, J.P. and Orkin, S.H.\",\"doi\":\"10.1016/S1097-2765(00)80312-3\",\"journal\":\"Molecular Cell\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Use of altered specificity mutants to probe a specific protein-protein interaction in differentiation: The GATA-1:FOG complex},\\n type = {article},\\n year = {1999},\\n pages = {219-228},\\n volume = {3},\\n id = {ed2ce0ef-2974-32de-ac4b-b652a7d61a3b},\\n created = {2023-01-10T01:46:46.200Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:46.200Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {GATA-1 and FOG (Friend of GATA-1) are each essential for erythroid and megakaryocyte development. FOG, a zinc finger protein, interacts with the amino (N) finger of GATA-1 and cooperates with GATA-1 to promote differentiation. To determine whether this interaction is critical for GATA- 1 action, we selected GATA-1 mutants in yeast that fail to interact with FOG but retain normal DNA binding, as well a compensatory FOG mutant that restores interaction. These novel GATA-1 mutants do not promote erythroid differentiation of GATA-1- erythroid cells. Differentiation is rescued by the second-site FOG mutant. Thus, interaction of FOG with GATA-1 is essential for the function of GATA-1 in erythroid differentiation. These findings provide a paradigm for dissecting protein-protein associations involved in mammalian development.},\\n bibtype = {article},\\n author = {Crispino, J.D. and Lodish, M.B. and MacKay, J.P. and Orkin, S.H.},\\n doi = {10.1016/S1097-2765(00)80312-3},\\n journal = {Molecular Cell},\\n number = {2}\\n}\",\"author_short\":[\"Crispino,  J.\",\"Lodish,  M.\",\"MacKay,  J.\",\"Orkin,  S.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"crispino-lodish-mackay-orkin-useofalteredspecificitymutantstoprobeaspecificproteinproteininteractionindifferentiationthegata1fogcomplex-1999\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The C-terminal region of the stalk domain of ubiquitous human kinesin heavy chain contains the binding site for kinesin light chain\",\"type\":\"article\",\"year\":\"1998\",\"pages\":\"16663-16670\",\"volume\":\"37\",\"id\":\"b73ee519-2d24-3501-8e20-4c8b4ead145e\",\"created\":\"2023-01-10T01:46:47.102Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:47.102Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The motor protein kinesin is a heterotetramer composed of two heavy chains of ~120 kDa and two light chains of ~6 kDa protein. Kinesin motor activity is dependent on the presence of ATP and microtubules. The kinesin light chain-binding site in human kinesin heavy chain was determined by reconstituting in vitro a complex of recombinant heavy and light chains. The proteins expressed in bacteria included oligohistidine-tagged fragments of human ubiquitous kinesin heavy chain, spanning most of the stalk and all of the tail domain (amino acids 555-963); and untagged, essentially full-length human kinesin light chain (4-569) along with N-terminal (4-363)and C-terminal (364-569) light chain fragments. Heavy chain fragments were attached to Ni2+-charged beads and incubated with untagged light chain fragments. Analysis of eluted complexes by SDS-PAGE and immunoblotting mapped the light chain-binding site in heavy chain to amino acids 771-813, a region close to the C-terminal end of the heavy chain stalk domain. In addition, only the full-length and N-terminal kinesin light chain fragments bound to this heavy chain region. Within this heavy chain region are four highly conserved contiguous heptad repeats (775-802) which are predicted to form a tight α- helical coiled-coil interaction with the heptad repeat-containing N-terminus of the light chain, in particular region 106-152 of human light chain. This predicted hydrophobic, α-helical coiled-coil interaction is supported by both circular dichroism spectroscopy of the recombinant kinesin heavy chain fragment 771-963, which displays an α-helical content of 70%, and the resistance of the heavy/light chain interdiction to high salt (0.5 M).\",\"bibtype\":\"article\",\"author\":\"Diefenbach, R.J. and Mackay, J.P. and Armati, P.J. and Cunningham, A.L.\",\"doi\":\"10.1021/bi981163r\",\"journal\":\"Biochemistry\",\"number\":\"47\",\"bibtex\":\"@article{\\n title = {The C-terminal region of the stalk domain of ubiquitous human kinesin heavy chain contains the binding site for kinesin light chain},\\n type = {article},\\n year = {1998},\\n pages = {16663-16670},\\n volume = {37},\\n id = {b73ee519-2d24-3501-8e20-4c8b4ead145e},\\n created = {2023-01-10T01:46:47.102Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:47.102Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The motor protein kinesin is a heterotetramer composed of two heavy chains of ~120 kDa and two light chains of ~6 kDa protein. Kinesin motor activity is dependent on the presence of ATP and microtubules. The kinesin light chain-binding site in human kinesin heavy chain was determined by reconstituting in vitro a complex of recombinant heavy and light chains. The proteins expressed in bacteria included oligohistidine-tagged fragments of human ubiquitous kinesin heavy chain, spanning most of the stalk and all of the tail domain (amino acids 555-963); and untagged, essentially full-length human kinesin light chain (4-569) along with N-terminal (4-363)and C-terminal (364-569) light chain fragments. Heavy chain fragments were attached to Ni2+-charged beads and incubated with untagged light chain fragments. Analysis of eluted complexes by SDS-PAGE and immunoblotting mapped the light chain-binding site in heavy chain to amino acids 771-813, a region close to the C-terminal end of the heavy chain stalk domain. In addition, only the full-length and N-terminal kinesin light chain fragments bound to this heavy chain region. Within this heavy chain region are four highly conserved contiguous heptad repeats (775-802) which are predicted to form a tight α- helical coiled-coil interaction with the heptad repeat-containing N-terminus of the light chain, in particular region 106-152 of human light chain. This predicted hydrophobic, α-helical coiled-coil interaction is supported by both circular dichroism spectroscopy of the recombinant kinesin heavy chain fragment 771-963, which displays an α-helical content of 70%, and the resistance of the heavy/light chain interdiction to high salt (0.5 M).},\\n bibtype = {article},\\n author = {Diefenbach, R.J. and Mackay, J.P. and Armati, P.J. and Cunningham, A.L.},\\n doi = {10.1021/bi981163r},\\n journal = {Biochemistry},\\n number = {47}\\n}\",\"author_short\":[\"Diefenbach,  R.\",\"Mackay,  J.\",\"Armati,  P.\",\"Cunningham,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"diefenbach-mackay-armati-cunningham-thecterminalregionofthestalkdomainofubiquitoushumankinesinheavychaincontainsthebindingsiteforkinesinlightchain-1998\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Key residues characteristic of GATA N-fingers are recognized by FOG\",\"type\":\"article\",\"year\":\"1998\",\"pages\":\"33595-33603\",\"volume\":\"273\",\"id\":\"e9e15c4c-0ebf-3abb-b74a-16640859e1ae\",\"created\":\"2023-01-10T01:46:48.015Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:48.015Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Protein-protein interactions play significant roles in the control of gene expression. These interactions often occur between small, discrete domains within different transcription factors. In particular, zinc fingers, usually regarded as DNA-binding domains, are now also known to be involved in mediating contacts between proteins. We have investigated the interaction between the erythroid transcription factor GATA-1 and its partner, the 9 zinc finger protein, FOG (Friend Of GATA). We demonstrate that this interaction represents a genuine finger-finger contact, which is dependent on zinc- coordinating residues within each protein. We map the contact domains to the core of the N-terminal zinc finger of GATA-1 and the 6th zinc finger of FOG. Using a scanning substitution strategy we identify key residues within the GATA-1 N-finger which are required for FOG binding. These residues are conserved in the N-fingers of all GATA proteins known to bind FOG, but are not found in the respective C-fingers. This observation may, therefore, account for the particular specificity of FOG for N-fingers. Interestingly, the key N-finger residues are seen to form a contiguous surface, when mapped onto the structure of the N-finger of GATA-1.\",\"bibtype\":\"article\",\"author\":\"Fox, A.H. and Kowalski, K. and King, G.F. and Mackay, J.P. and Crossley, M.\",\"doi\":\"10.1074/jbc.273.50.33595\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"50\",\"bibtex\":\"@article{\\n title = {Key residues characteristic of GATA N-fingers are recognized by FOG},\\n type = {article},\\n year = {1998},\\n pages = {33595-33603},\\n volume = {273},\\n id = {e9e15c4c-0ebf-3abb-b74a-16640859e1ae},\\n created = {2023-01-10T01:46:48.015Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:48.015Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Protein-protein interactions play significant roles in the control of gene expression. These interactions often occur between small, discrete domains within different transcription factors. In particular, zinc fingers, usually regarded as DNA-binding domains, are now also known to be involved in mediating contacts between proteins. We have investigated the interaction between the erythroid transcription factor GATA-1 and its partner, the 9 zinc finger protein, FOG (Friend Of GATA). We demonstrate that this interaction represents a genuine finger-finger contact, which is dependent on zinc- coordinating residues within each protein. We map the contact domains to the core of the N-terminal zinc finger of GATA-1 and the 6th zinc finger of FOG. Using a scanning substitution strategy we identify key residues within the GATA-1 N-finger which are required for FOG binding. These residues are conserved in the N-fingers of all GATA proteins known to bind FOG, but are not found in the respective C-fingers. This observation may, therefore, account for the particular specificity of FOG for N-fingers. Interestingly, the key N-finger residues are seen to form a contiguous surface, when mapped onto the structure of the N-finger of GATA-1.},\\n bibtype = {article},\\n author = {Fox, A.H. and Kowalski, K. and King, G.F. and Mackay, J.P. and Crossley, M.},\\n doi = {10.1074/jbc.273.50.33595},\\n journal = {Journal of Biological Chemistry},\\n number = {50}\\n}\",\"author_short\":[\"Fox,  A.\",\"Kowalski,  K.\",\"King,  G.\",\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"fox-kowalski-king-mackay-crossley-keyresiduescharacteristicofgatanfingersarerecognizedbyfog-1998\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Zinc fingers are sticking together\",\"type\":\"article\",\"year\":\"1998\",\"pages\":\"1-4\",\"volume\":\"23\",\"id\":\"76060f13-2fda-31b9-b159-68b649adeb20\",\"created\":\"2023-01-10T01:46:48.915Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:48.915Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Crossley, M.\",\"doi\":\"10.1016/S0968-0004(97)01168-7\",\"journal\":\"Trends in Biochemical Sciences\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Zinc fingers are sticking together},\\n type = {article},\\n year = {1998},\\n pages = {1-4},\\n volume = {23},\\n id = {76060f13-2fda-31b9-b159-68b649adeb20},\\n created = {2023-01-10T01:46:48.915Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:48.915Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Mackay, J.P. and Crossley, M.},\\n doi = {10.1016/S0968-0004(97)01168-7},\\n journal = {Trends in Biochemical Sciences},\\n number = {1}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-crossley-zincfingersarestickingtogether-1998\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Interactions of the Antitumor Agent Molybdocene Dichloride with Oligonucleotides\",\"type\":\"article\",\"year\":\"1998\",\"pages\":\"2432-2437\",\"volume\":\"37\",\"id\":\"3622d446-6dc4-3dca-890e-5982c68ebdd0\",\"created\":\"2023-01-10T01:46:49.823Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:49.823Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The interactions between the antitumor-active metallocene molybdocene dichloride (Cp2MoCl2) and four oligonucleotides have been studied by 1H and 31P NMR spectroscopy. In 50 mM salt solutions of molybdocene dichloride, hydrolysis of the halide ligands occurs to give a solution with pD 2, containing a species in which both Cp rings remain metal bound for 24 h. At pD 7, however, partial hydrolysis of the Cp rings (∼30%) occurs after 24 h. Addition of an aqueous solution of molybdocene dichloride in 50 mM salt to the self-complementary sequence d(CGCATATGCG)2, maintaining the pD at 6.0-7.0, showed no evidence for the formation of a metallocene-oligonucleotide complex, and only peaks arising from hydrolysis of molybdocene dichloride were detected. A similar result was obtained in titration experiments with the single-stranded sequence d(ATGGTA) at pD 6.5-7.0. However, at pD 3.0, new signals assigned to a molybdocene-oligonuleotide complex or complexes were detected in the 1H NMR spectrum. No change was observed in the 31P NMR spectrum. The complex or complexes formed between molybdocene dichloride and d(ATGGTA) are stable for hours at pD 3.0; at higher pD, the complex is destabilized and only peaks arising from hydrolysis of molybdocene dichloride are detected. Titration experiments at low pD with the dinucleotide dCpG showed a new set of signals in the 1H NMR spectrum, tentatively assigned to formation of a complex arising due to coordination of molybdenum to guanine N7 and/or cytosine N3. At pD 7.0, these signals disappeared. The results obtained show that stable oligonucleotide adducts are not formed in 50 mM salt at pD 7.0, and hence it is highly unlikely that formation of molybdocene - DNA adducts in vivo is the primary action that is responsible for the antitumor properties of molybdocene dichloride.\",\"bibtype\":\"article\",\"author\":\"Harding, M.M. and Mokdsi, G. and Mackay, J.P. and Prodigalidad, M. and Lucas, S.W.\",\"doi\":\"10.1021/ic971205k\",\"journal\":\"Inorganic Chemistry\",\"number\":\"10\",\"bibtex\":\"@article{\\n title = {Interactions of the Antitumor Agent Molybdocene Dichloride with Oligonucleotides},\\n type = {article},\\n year = {1998},\\n pages = {2432-2437},\\n volume = {37},\\n id = {3622d446-6dc4-3dca-890e-5982c68ebdd0},\\n created = {2023-01-10T01:46:49.823Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:49.823Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The interactions between the antitumor-active metallocene molybdocene dichloride (Cp2MoCl2) and four oligonucleotides have been studied by 1H and 31P NMR spectroscopy. In 50 mM salt solutions of molybdocene dichloride, hydrolysis of the halide ligands occurs to give a solution with pD 2, containing a species in which both Cp rings remain metal bound for 24 h. At pD 7, however, partial hydrolysis of the Cp rings (∼30%) occurs after 24 h. Addition of an aqueous solution of molybdocene dichloride in 50 mM salt to the self-complementary sequence d(CGCATATGCG)2, maintaining the pD at 6.0-7.0, showed no evidence for the formation of a metallocene-oligonucleotide complex, and only peaks arising from hydrolysis of molybdocene dichloride were detected. A similar result was obtained in titration experiments with the single-stranded sequence d(ATGGTA) at pD 6.5-7.0. However, at pD 3.0, new signals assigned to a molybdocene-oligonuleotide complex or complexes were detected in the 1H NMR spectrum. No change was observed in the 31P NMR spectrum. The complex or complexes formed between molybdocene dichloride and d(ATGGTA) are stable for hours at pD 3.0; at higher pD, the complex is destabilized and only peaks arising from hydrolysis of molybdocene dichloride are detected. Titration experiments at low pD with the dinucleotide dCpG showed a new set of signals in the 1H NMR spectrum, tentatively assigned to formation of a complex arising due to coordination of molybdenum to guanine N7 and/or cytosine N3. At pD 7.0, these signals disappeared. The results obtained show that stable oligonucleotide adducts are not formed in 50 mM salt at pD 7.0, and hence it is highly unlikely that formation of molybdocene - DNA adducts in vivo is the primary action that is responsible for the antitumor properties of molybdocene dichloride.},\\n bibtype = {article},\\n author = {Harding, M.M. and Mokdsi, G. and Mackay, J.P. and Prodigalidad, M. and Lucas, S.W.},\\n doi = {10.1021/ic971205k},\\n journal = {Inorganic Chemistry},\\n number = {10}\\n}\",\"author_short\":[\"Harding,  M.\",\"Mokdsi,  G.\",\"Mackay,  J.\",\"Prodigalidad,  M.\",\"Lucas,  S.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"harding-mokdsi-mackay-prodigalidad-lucas-interactionsoftheantitumoragentmolybdocenedichloridewitholigonucleotides-1998\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Involvement of the N-finger in the self-association of GATA-1\",\"type\":\"article\",\"year\":\"1998\",\"pages\":\"30560-30567\",\"volume\":\"273\",\"id\":\"4c338f22-0421-34a2-92f2-d97f844ae7db\",\"created\":\"2023-01-10T01:46:50.749Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:50.749Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Zinc fingers are recognized as small protein domains that bind to specific DNA sequences. Recently however, zinc fingers from a number of proteins, in particular the GATA family of transcription factors, have also been implicated in specific protein-protein interactions. The erythroid protein GATA-1 contains two zinc fingers: the C-finger, which is sufficient for sequence-specific DNA-binding, and the N-finger, which appears both to modulate DNA-binding and to interact with other transcription factors. We have expressed and purified the N-finger domain and investigated its involvement in the self-association of GATA-1. We demonstrate that this domain does not homodimerize but instead makes intermolecular contacts with the C-finger, suggesting that GATA dimers are maintained by reciprocal N- finger-C-finger contacts. Deletion analysis identifies a 25-residue region, C-terminal to the core N-finger domain, that is sufficient for interaction with intact GATA-1. A similar subdomain exists C-terminal to the C-finger, and we show that self-association is substantially reduced when both subdomains are disrupted by mutation. Moreover, mutations that impair GATA-1 self-association also interfere with its ability to activate transcription in transfection studies.\",\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Kowalski, K. and Fox, A.H. and Czolij, R. and King, G.F. and Crossley, M.\",\"doi\":\"10.1074/jbc.273.46.30560\",\"journal\":\"Journal of Biological Chemistry\",\"number\":\"46\",\"bibtex\":\"@article{\\n title = {Involvement of the N-finger in the self-association of GATA-1},\\n type = {article},\\n year = {1998},\\n pages = {30560-30567},\\n volume = {273},\\n id = {4c338f22-0421-34a2-92f2-d97f844ae7db},\\n created = {2023-01-10T01:46:50.749Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:50.749Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Zinc fingers are recognized as small protein domains that bind to specific DNA sequences. Recently however, zinc fingers from a number of proteins, in particular the GATA family of transcription factors, have also been implicated in specific protein-protein interactions. The erythroid protein GATA-1 contains two zinc fingers: the C-finger, which is sufficient for sequence-specific DNA-binding, and the N-finger, which appears both to modulate DNA-binding and to interact with other transcription factors. We have expressed and purified the N-finger domain and investigated its involvement in the self-association of GATA-1. We demonstrate that this domain does not homodimerize but instead makes intermolecular contacts with the C-finger, suggesting that GATA dimers are maintained by reciprocal N- finger-C-finger contacts. Deletion analysis identifies a 25-residue region, C-terminal to the core N-finger domain, that is sufficient for interaction with intact GATA-1. A similar subdomain exists C-terminal to the C-finger, and we show that self-association is substantially reduced when both subdomains are disrupted by mutation. Moreover, mutations that impair GATA-1 self-association also interfere with its ability to activate transcription in transfection studies.},\\n bibtype = {article},\\n author = {Mackay, J.P. and Kowalski, K. and Fox, A.H. and Czolij, R. and King, G.F. and Crossley, M.},\\n doi = {10.1074/jbc.273.46.30560},\\n journal = {Journal of Biological Chemistry},\\n number = {46}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Kowalski,  K.\",\"Fox,  A.\",\"Czolij,  R.\",\"King,  G.\",\"Crossley,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-kowalski-fox-czolij-king-crossley-involvementofthenfingerintheselfassociationofgata1-1998\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides\",\"type\":\"article\",\"year\":\"1997\",\"pages\":\"67-77\",\"volume\":\"4\",\"id\":\"e3e2a974-5c9b-3e6e-8d6b-ec39b861bc0a\",\"created\":\"2023-01-10T01:46:51.663Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:51.663Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides. © 1997 ESCOM Science Publishers B.V.\",\"bibtype\":\"article\",\"author\":\"Bains, N.P.S. and Wilce, J.A. and Heuer, K.H. and Tunstall, M. and Mackay, J.P. and Bennett, M.R. and Weiss, A.S. and King, G.F.\",\"doi\":\"10.1007/BF02443517\",\"journal\":\"International Journal of Peptide Research and Therapeutics\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides},\\n type = {article},\\n year = {1997},\\n pages = {67-77},\\n volume = {4},\\n id = {e3e2a974-5c9b-3e6e-8d6b-ec39b861bc0a},\\n created = {2023-01-10T01:46:51.663Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:51.663Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides. © 1997 ESCOM Science Publishers B.V.},\\n bibtype = {article},\\n author = {Bains, N.P.S. and Wilce, J.A. and Heuer, K.H. and Tunstall, M. and Mackay, J.P. and Bennett, M.R. and Weiss, A.S. and King, G.F.},\\n doi = {10.1007/BF02443517},\\n journal = {International Journal of Peptide Research and Therapeutics},\\n number = {2}\\n}\",\"author_short\":[\"Bains,  N.\",\"Wilce,  J.\",\"Heuer,  K.\",\"Tunstall,  M.\",\"Mackay,  J.\",\"Bennett,  M.\",\"Weiss,  A.\",\"King,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR\",\"type\":\"article\",\"year\":\"1997\",\"pages\":\"1-8\",\"volume\":\"10\",\"id\":\"53beac46-6091-3dfa-988a-801d2a1ac05f\",\"created\":\"2023-01-10T01:46:52.607Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:52.607Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.\",\"bibtype\":\"article\",\"author\":\"Dingley, A.J. and Mackay, J.P. and Shaw, G.L. and Hambly, B.D. and King, G.F.\",\"doi\":\"10.1023/A:1018339526108\",\"journal\":\"Journal of Biomolecular NMR\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR},\\n type = {article},\\n year = {1997},\\n pages = {1-8},\\n volume = {10},\\n id = {53beac46-6091-3dfa-988a-801d2a1ac05f},\\n created = {2023-01-10T01:46:52.607Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:52.607Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.},\\n bibtype = {article},\\n author = {Dingley, A.J. and Mackay, J.P. and Shaw, G.L. and Hambly, B.D. and King, G.F.},\\n doi = {10.1023/A:1018339526108},\\n journal = {Journal of Biomolecular NMR},\\n number = {1}\\n}\",\"author_short\":[\"Dingley,  A.\",\"Mackay,  J.\",\"Shaw,  G.\",\"Hambly,  B.\",\"King,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"The structure of versutoxin (δ-atracotoxin-Hv1) provides insights into the binding of site 3 neurotoxins to the voltage-gated sodium channel\",\"type\":\"article\",\"year\":\"1997\",\"pages\":\"1525-1535\",\"volume\":\"5\",\"id\":\"c641551b-c512-3935-9b0e-32d0d876b55e\",\"created\":\"2023-01-10T01:46:53.511Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:53.511Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background: Versutoxin (δ-ACTX-Hv1) is the major component of the venom of the Australian Blue Mountains funnel web spider, Hadronyche versuta. δ-ACTX-Hv1 produces potentially fatal neurotoxic symptoms in primates by slowing the inactivation of voltage-gated sodium channels; δ-ACTX-Hv1 is therefore a useful tool for studying sodium channel function. We have determined the three-dimensional structure of δ-ACTX-Hv1 as the first step towards understanding the molecular basis of its interaction with these channels. Results: The solution structure of δ-ACTX-Hv1, determined using NMR spectroscopy, comprises a core β region containing a triple-stranded antiparallel β sheet, a thumb-like extension protruding from the β region and a C-terminal 310 helix that is appended to the β domain by virtue of a disulphide bond. The β region contains a cystine knot motif similar to that seen in other neurotoxic polypeptides. The structure shows homology with μ-agatoxin-I, a spider toxin that also modifies the inactivation kinetics of vertebrate voltage-gated sodium channels. More surprisingly, δ-ACTX-Hv1 shows both sequence and structural homology with gurmarin, a plant polypeptide. This similarity leads us to suggest that the sweet-taste suppression elicited by gurmarin may result from an interaction with one of the downstream ion channels involved in sweet-taste transduction. Conclusions: δ-ACTX-Hv1 shows no structural homology with either sea anemone or α-scorpion toxins, both of which also modify the inactivation kinetics of voltage-gated sodium channels by interacting with channel recognition site 3. However, we have shown that δ-ACTX-Hv1 contains charged residues that are topologically related to those implicated in the binding of sea anemone and α-scorpion toxins to mammalian voltage-gated sodium channels, suggesting similarities in their mode of interaction with these channels.\",\"bibtype\":\"article\",\"author\":\"Fletcher, J.I. and Chapman, B.E. and Mackay, J.P. and Howden, M.E.H. and King, G.F.\",\"doi\":\"10.1016/S0969-2126(97)00301-8\",\"journal\":\"Structure\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {The structure of versutoxin (δ-atracotoxin-Hv1) provides insights into the binding of site 3 neurotoxins to the voltage-gated sodium channel},\\n type = {article},\\n year = {1997},\\n pages = {1525-1535},\\n volume = {5},\\n id = {c641551b-c512-3935-9b0e-32d0d876b55e},\\n created = {2023-01-10T01:46:53.511Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:53.511Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background: Versutoxin (δ-ACTX-Hv1) is the major component of the venom of the Australian Blue Mountains funnel web spider, Hadronyche versuta. δ-ACTX-Hv1 produces potentially fatal neurotoxic symptoms in primates by slowing the inactivation of voltage-gated sodium channels; δ-ACTX-Hv1 is therefore a useful tool for studying sodium channel function. We have determined the three-dimensional structure of δ-ACTX-Hv1 as the first step towards understanding the molecular basis of its interaction with these channels. Results: The solution structure of δ-ACTX-Hv1, determined using NMR spectroscopy, comprises a core β region containing a triple-stranded antiparallel β sheet, a thumb-like extension protruding from the β region and a C-terminal 310 helix that is appended to the β domain by virtue of a disulphide bond. The β region contains a cystine knot motif similar to that seen in other neurotoxic polypeptides. The structure shows homology with μ-agatoxin-I, a spider toxin that also modifies the inactivation kinetics of vertebrate voltage-gated sodium channels. More surprisingly, δ-ACTX-Hv1 shows both sequence and structural homology with gurmarin, a plant polypeptide. This similarity leads us to suggest that the sweet-taste suppression elicited by gurmarin may result from an interaction with one of the downstream ion channels involved in sweet-taste transduction. Conclusions: δ-ACTX-Hv1 shows no structural homology with either sea anemone or α-scorpion toxins, both of which also modify the inactivation kinetics of voltage-gated sodium channels by interacting with channel recognition site 3. However, we have shown that δ-ACTX-Hv1 contains charged residues that are topologically related to those implicated in the binding of sea anemone and α-scorpion toxins to mammalian voltage-gated sodium channels, suggesting similarities in their mode of interaction with these channels.},\\n bibtype = {article},\\n author = {Fletcher, J.I. and Chapman, B.E. and Mackay, J.P. and Howden, M.E.H. and King, G.F.},\\n doi = {10.1016/S0969-2126(97)00301-8},\\n journal = {Structure},\\n number = {11}\\n}\",\"author_short\":[\"Fletcher,  J.\",\"Chapman,  B.\",\"Mackay,  J.\",\"Howden,  M.\",\"King,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"fletcher-chapman-mackay-howden-king-thestructureofversutoxinatracotoxinhv1providesinsightsintothebindingofsite3neurotoxinstothevoltagegatedsodiumchannel-1997\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides\",\"type\":\"article\",\"year\":\"1997\",\"pages\":\"67-77\",\"volume\":\"4\",\"id\":\"424daf6d-00c1-3bcf-ad60-9abdb95c2e4b\",\"created\":\"2023-01-10T01:46:54.417Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:54.417Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides.\",\"bibtype\":\"article\",\"author\":\"Bains, N.P.S. and Wilce, J.A. and Heuer, K.H. and Tunstall, M. and Mackay, J.P. and Bennett, M.R. and Weiss, A.S. and King, G.F.\",\"doi\":\"10.1007/BF02443517\",\"journal\":\"Letters in Peptide Science\",\"number\":\"2\",\"bibtex\":\"@article{\\n title = {Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides},\\n type = {article},\\n year = {1997},\\n pages = {67-77},\\n volume = {4},\\n id = {424daf6d-00c1-3bcf-ad60-9abdb95c2e4b},\\n created = {2023-01-10T01:46:54.417Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:54.417Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides.},\\n bibtype = {article},\\n author = {Bains, N.P.S. and Wilce, J.A. and Heuer, K.H. and Tunstall, M. and Mackay, J.P. and Bennett, M.R. and Weiss, A.S. and King, G.F.},\\n doi = {10.1007/BF02443517},\\n journal = {Letters in Peptide Science},\\n number = {2}\\n}\",\"author_short\":[\"Bains,  N.\",\"Wilce,  J.\",\"Heuer,  K.\",\"Tunstall,  M.\",\"Mackay,  J.\",\"Bennett,  M.\",\"Weiss,  A.\",\"King,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"DMA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II\",\"type\":\"article\",\"year\":\"1997\",\"pages\":\"387-398\",\"volume\":\"42\",\"id\":\"964a530f-b34f-3e6c-917f-3ea08bb03126\",\"created\":\"2023-01-10T01:46:55.338Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:55.338Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley & Sons, Inc.\",\"bibtype\":\"article\",\"author\":\"Harding, M.M. and Krippner, G.Y. and Shelton, C.J. and Rodger, A. and Sanders, K.J. and Mackay, J.P. and Prakash, A.S.\",\"doi\":\"10.1002/(SICI)1097-0282(19971005)42:4<387::AID-BIP2>3.0.CO;2-M\",\"journal\":\"Biopolymers - Nucleic Acid Sciences Section\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {DMA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II},\\n type = {article},\\n year = {1997},\\n pages = {387-398},\\n volume = {42},\\n id = {964a530f-b34f-3e6c-917f-3ea08bb03126},\\n created = {2023-01-10T01:46:55.338Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:55.338Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley & Sons, Inc.},\\n bibtype = {article},\\n author = {Harding, M.M. and Krippner, G.Y. and Shelton, C.J. and Rodger, A. and Sanders, K.J. and Mackay, J.P. and Prakash, A.S.},\\n doi = {10.1002/(SICI)1097-0282(19971005)42:4<387::AID-BIP2>3.0.CO;2-M},\\n journal = {Biopolymers - Nucleic Acid Sciences Section},\\n number = {4}\\n}\",\"author_short\":[\"Harding,  M.\",\"Krippner,  G.\",\"Shelton,  C.\",\"Rodger,  A.\",\"Sanders,  K.\",\"Mackay,  J.\",\"Prakash,  A.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"harding-krippner-shelton-rodger-sanders-mackay-prakash-dmabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Δ-cis-α-[Ru(RR-picchxnMe<inf>2</inf>)(phen)]<sup>2+</sup> shows minor groove AT selectivity with oligonucleotides\",\"type\":\"article\",\"year\":\"1997\",\"pages\":\"1623-1624\",\"id\":\"9b3e2289-c499-39c6-9a25-43e57803b26c\",\"created\":\"2023-01-10T01:46:56.245Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:56.245Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"NMR studies show that the ternary octahedral Δ-cis-α-[Ru(RR-picchxnMe2)(phen)]2+ cation binds with AT selectivity in the minor groove of [d(CGCGATCGCG)2] and [d(ATATCGATAT)2] duplexes through a non-intercalative interaction.\",\"bibtype\":\"article\",\"author\":\"Proudfoot, E.M. and Mackay, J.P. and Vagg, R.S. and Vickery, K.A. and Williams, P.A. and Karuso, P.\",\"doi\":\"10.1039/a704001f\",\"journal\":\"Chemical Communications\",\"number\":\"17\",\"bibtex\":\"@article{\\n title = {Δ-cis-α-[Ru(RR-picchxnMe<inf>2</inf>)(phen)]<sup>2+</sup> shows minor groove AT selectivity with oligonucleotides},\\n type = {article},\\n year = {1997},\\n pages = {1623-1624},\\n id = {9b3e2289-c499-39c6-9a25-43e57803b26c},\\n created = {2023-01-10T01:46:56.245Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:56.245Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {NMR studies show that the ternary octahedral Δ-cis-α-[Ru(RR-picchxnMe2)(phen)]2+ cation binds with AT selectivity in the minor groove of [d(CGCGATCGCG)2] and [d(ATATCGATAT)2] duplexes through a non-intercalative interaction.},\\n bibtype = {article},\\n author = {Proudfoot, E.M. and Mackay, J.P. and Vagg, R.S. and Vickery, K.A. and Williams, P.A. and Karuso, P.},\\n doi = {10.1039/a704001f},\\n journal = {Chemical Communications},\\n number = {17}\\n}\",\"author_short\":[\"Proudfoot,  E.\",\"Mackay,  J.\",\"Vagg,  R.\",\"Vickery,  K.\",\"Williams,  P.\",\"Karuso,  P.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"proudfoot-mackay-vagg-vickery-williams-karuso-cisrurrpicchxnmeinf2infphensup2supshowsminorgrooveatselectivitywitholigonucleotides-1997\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Assignment of the <sup>1</sup>H NMR spectrum and solution conformation of the antitumour antibiotic ditrisarubicin B\",\"type\":\"article\",\"year\":\"1996\",\"pages\":\"5617-5624\",\"volume\":\"52\",\"id\":\"47129b2c-d23a-3485-8fba-32923cad98a3\",\"created\":\"2023-01-10T01:46:57.152Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:57.152Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Complete assignment of the 1H NMR spectrum of ditrisarubicin B, a member of the anthracycline antitumour antibiotics, in acetonitrile is reported. The two trisaccharide chains are highly structured and their conformation supports their role as preorganised DNA minor groove binders which bind to DNA on intercalation of the tetracyclic chromophore.\",\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Shelton, C.J. and Harding, M.M.\",\"doi\":\"10.1016/0040-4020(96)00198-6\",\"journal\":\"Tetrahedron\",\"number\":\"15\",\"bibtex\":\"@article{\\n title = {Assignment of the <sup>1</sup>H NMR spectrum and solution conformation of the antitumour antibiotic ditrisarubicin B},\\n type = {article},\\n year = {1996},\\n pages = {5617-5624},\\n volume = {52},\\n id = {47129b2c-d23a-3485-8fba-32923cad98a3},\\n created = {2023-01-10T01:46:57.152Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:57.152Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Complete assignment of the 1H NMR spectrum of ditrisarubicin B, a member of the anthracycline antitumour antibiotics, in acetonitrile is reported. The two trisaccharide chains are highly structured and their conformation supports their role as preorganised DNA minor groove binders which bind to DNA on intercalation of the tetracyclic chromophore.},\\n bibtype = {article},\\n author = {Mackay, J.P. and Shelton, C.J. and Harding, M.M.},\\n doi = {10.1016/0040-4020(96)00198-6},\\n journal = {Tetrahedron},\\n number = {15}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Shelton,  C.\",\"Harding,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-shelton-harding-assignmentofthesup1suphnmrspectrumandsolutionconformationoftheantitumourantibioticditrisarubicinb-1996\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Backbone dynamics of the c-Jun leucine zipper: <sup>15</sup>N NMR relaxation studies\",\"type\":\"article\",\"year\":\"1996\",\"pages\":\"4867-4877\",\"volume\":\"35\",\"id\":\"362dbcd0-e82e-357a-a5d5-97ec3d2ac62e\",\"created\":\"2023-01-10T01:46:58.102Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:58.102Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"The backbone dynamics of the coiled-coil leucine zipper domain of c-Jun have been studied using proton-detected two-dimensional 1H-15N NMR spectroscopy. Longitudinal (T1) and transverse (T2) 15N relaxation times, together with 1H15N NOEs, were measured and analyzed by considering the protein to approximate a prolate ellipsoid. An analysis of the T1/T2 ratios for residues in the well-structured section of the protein showed that a model for the spectral density function in which the protein is considered to reorient anisotropically fitted the data significantly better than an isotropic model. Order parameters (S2) in the range 0.7-0.9 were observed for most residues, with lower values near the C-terminus, consistent with fraying of the two helices comprising the coiled-coil. Because nearly all of the N-H vectors have small angles to the long axis of the molecule, there was some uncertainty in the value of the rotational diffusion coefficient Dpar, which describes rotation about the long axis. Thus, an alternative method was examined for its ability to provide independent estimates of Dpar and Dperp (the diffusion coefficient describing rotation about axes perpendicular to the long axis); the translational diffusion coefficient (Dt) of the protein was measured, and hydrodynamic calculations were used to predict Dpar and Dperp. However, the derived rotational diffusion coefficients proved to be very dependent on the hydrodynamic model used to relate Dt to Dpar and Dperp, and consequently the values obtained from the T1/T2 analysis were used in the order-parameter analysis. Although it has previously been reported that the side chain of a polar residue at the dimer interface, Asn22, undergoes a conformational exchange process and destabilizes the dimer, no evidence of increased backbone mobility in this region was detected, suggesting that this process is confined to the Asn side chain.\",\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Shaw, G.L. and King, G.F.\",\"doi\":\"10.1021/bi952761y\",\"journal\":\"Biochemistry\",\"number\":\"15\",\"bibtex\":\"@article{\\n title = {Backbone dynamics of the c-Jun leucine zipper: <sup>15</sup>N NMR relaxation studies},\\n type = {article},\\n year = {1996},\\n pages = {4867-4877},\\n volume = {35},\\n id = {362dbcd0-e82e-357a-a5d5-97ec3d2ac62e},\\n created = {2023-01-10T01:46:58.102Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:58.102Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {The backbone dynamics of the coiled-coil leucine zipper domain of c-Jun have been studied using proton-detected two-dimensional 1H-15N NMR spectroscopy. Longitudinal (T1) and transverse (T2) 15N relaxation times, together with 1H15N NOEs, were measured and analyzed by considering the protein to approximate a prolate ellipsoid. An analysis of the T1/T2 ratios for residues in the well-structured section of the protein showed that a model for the spectral density function in which the protein is considered to reorient anisotropically fitted the data significantly better than an isotropic model. Order parameters (S2) in the range 0.7-0.9 were observed for most residues, with lower values near the C-terminus, consistent with fraying of the two helices comprising the coiled-coil. Because nearly all of the N-H vectors have small angles to the long axis of the molecule, there was some uncertainty in the value of the rotational diffusion coefficient Dpar, which describes rotation about the long axis. Thus, an alternative method was examined for its ability to provide independent estimates of Dpar and Dperp (the diffusion coefficient describing rotation about axes perpendicular to the long axis); the translational diffusion coefficient (Dt) of the protein was measured, and hydrodynamic calculations were used to predict Dpar and Dperp. However, the derived rotational diffusion coefficients proved to be very dependent on the hydrodynamic model used to relate Dt to Dpar and Dperp, and consequently the values obtained from the T1/T2 analysis were used in the order-parameter analysis. Although it has previously been reported that the side chain of a polar residue at the dimer interface, Asn22, undergoes a conformational exchange process and destabilizes the dimer, no evidence of increased backbone mobility in this region was detected, suggesting that this process is confined to the Asn side chain.},\\n bibtype = {article},\\n author = {Mackay, J.P. and Shaw, G.L. and King, G.F.},\\n doi = {10.1021/bi952761y},\\n journal = {Biochemistry},\\n number = {15}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Shaw,  G.\",\"King,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-shaw-king-backbonedynamicsofthecjunleucinezippersup15supnnmrrelaxationstudies-1996\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Development of a sensitive peptide-based immunoassay: Application to detection of the Jun and Fos oncoproteins\",\"type\":\"article\",\"year\":\"1996\",\"pages\":\"9069-9075\",\"volume\":\"35\",\"id\":\"3ec35428-2577-35f5-8d93-7aae09b3b3bc\",\"created\":\"2023-01-10T01:46:59.013Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:46:59.013Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"c-Jun and c-Fos belong to the bZIP class of transcriptional activator proteins, many of which have been implicated in the neoplastic transformation of cells. We are interested in engineering dominant-negative leucine zipper (LZ) peptides as a means of sequestering these proteins in vivo in order to suppress their transcriptional regulatory activity. Toward this end, we have developed a novel immunoassay for measuring the dimerization affinities of dimeric Jun and Fos complexes. This peptide-based ELISA relies on the fact that Jun and Fos preferentially form heterodimers via their leucine zipper domains. Recombinant Jun leucine zipper peptides (either native JunLZ or a V36→E point mutant) were labeled with biotin and specifically bound through a leucine zipper interaction to a FosLZ-glutathione S-transferase fusion protein adsorbed onto the wells of an ELISA tray. Jun:Fos complexes were subsequently detected using a recently developed streptavidin-based amplification system known as enzyme complex amplification [Wilson, M. R., and Easterbrook-Smith, S. B. (1993) Anal. Biochem. 209, 183-187]. This ELISA system can detect subnanomolar concentrations of Jun and Fos, thus allowing determination of the dissociation constants for complex formation. The dissociation constant for formation of the native JunLZ:FosLZ heterodimer at 37 °C was determined to be 0.99 ± 0.30 nM, while that for JunLZ(V36E):FosLZ heterodimer was 0.90 ± 0.13 μM. These results demonstrate that the novel peptide-based ELISA described herein is simple and sensitive and can be used to rapidly screen for potential dominant-negative leucine zipper peptides.\",\"bibtype\":\"article\",\"author\":\"Heuer, K.H. and Mackay, J.P. and Podzebenko, P. and Bains, N.P.S. and Weiss, A.S. and King, G.F. and Easterbrook-Smith, S.B.\",\"doi\":\"10.1021/bi952817o\",\"journal\":\"Biochemistry\",\"number\":\"28\",\"bibtex\":\"@article{\\n title = {Development of a sensitive peptide-based immunoassay: Application to detection of the Jun and Fos oncoproteins},\\n type = {article},\\n year = {1996},\\n pages = {9069-9075},\\n volume = {35},\\n id = {3ec35428-2577-35f5-8d93-7aae09b3b3bc},\\n created = {2023-01-10T01:46:59.013Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:46:59.013Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {c-Jun and c-Fos belong to the bZIP class of transcriptional activator proteins, many of which have been implicated in the neoplastic transformation of cells. We are interested in engineering dominant-negative leucine zipper (LZ) peptides as a means of sequestering these proteins in vivo in order to suppress their transcriptional regulatory activity. Toward this end, we have developed a novel immunoassay for measuring the dimerization affinities of dimeric Jun and Fos complexes. This peptide-based ELISA relies on the fact that Jun and Fos preferentially form heterodimers via their leucine zipper domains. Recombinant Jun leucine zipper peptides (either native JunLZ or a V36→E point mutant) were labeled with biotin and specifically bound through a leucine zipper interaction to a FosLZ-glutathione S-transferase fusion protein adsorbed onto the wells of an ELISA tray. Jun:Fos complexes were subsequently detected using a recently developed streptavidin-based amplification system known as enzyme complex amplification [Wilson, M. R., and Easterbrook-Smith, S. B. (1993) Anal. Biochem. 209, 183-187]. This ELISA system can detect subnanomolar concentrations of Jun and Fos, thus allowing determination of the dissociation constants for complex formation. The dissociation constant for formation of the native JunLZ:FosLZ heterodimer at 37 °C was determined to be 0.99 ± 0.30 nM, while that for JunLZ(V36E):FosLZ heterodimer was 0.90 ± 0.13 μM. These results demonstrate that the novel peptide-based ELISA described herein is simple and sensitive and can be used to rapidly screen for potential dominant-negative leucine zipper peptides.},\\n bibtype = {article},\\n author = {Heuer, K.H. and Mackay, J.P. and Podzebenko, P. and Bains, N.P.S. and Weiss, A.S. and King, G.F. and Easterbrook-Smith, S.B.},\\n doi = {10.1021/bi952817o},\\n journal = {Biochemistry},\\n number = {28}\\n}\",\"author_short\":[\"Heuer,  K.\",\"Mackay,  J.\",\"Podzebenko,  P.\",\"Bains,  N.\",\"Weiss,  A.\",\"King,  G.\",\"Easterbrook-Smith,  S.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"heuer-mackay-podzebenko-bains-weiss-king-easterbrooksmith-developmentofasensitivepeptidebasedimmunoassayapplicationtodetectionofthejunandfosoncoproteins-1996\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Measuring protein self-association using pulsed-field-gradient NMR spectroscopy: Application to myosin light chain 2\",\"type\":\"article\",\"year\":\"1995\",\"pages\":\"321-328\",\"volume\":\"6\",\"id\":\"a162714d-04d2-3f71-8080-eeb26343a53a\",\"created\":\"2023-01-10T01:47:00.198Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:47:00.198Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"At the millimolar concentrations required for structural studies, NMR spectra of the calcium-binding protein myosin light chain 2 (MLC2) showed resonance line widths indicative of extensive self-association. Pulsed-field-gradient (PFG) NMR spectroscopy was used to examine whether MLC2 aggregation could be prevented by the zwitterionic bile salt derivative 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). PFG NMR measurements indicated that CHAPS was capable of preventing MLC2 self-association, but only at concentrations well above the critical micelle concentration of ∼7.5 mM. CHAPS was most effective at a concentration of 22.5 mM, where the apparent molecular mass of MLC2 correponded to a protein monomer plus seven molecules of bound detergent. The resolution and sensitivity of 2D 15N-1H HSQC spectra of MLC2 were markedly improved by the addition of 25 mM CHAPS, consistent with a reduction in aggregation following addition of the detergent. The average amide nitrogen T2 value for MLC2 increased from ∼30 ms in the absence of CHAPS to ∼56 ms in the presence of 25 mM CHAPS. The results of this study lead us to propose that PFG NMR spectroscopy can be used as a facile alternative to conventional techniques such as analytical ultracentrifugation for examining the self-association of biological macromolecules. © 1995 ESCOM Science Publishers B.V.\",\"bibtype\":\"article\",\"author\":\"Dingley, A.J. and Mackay, J.P. and Chapman, B.E. and Morris, M.B. and Kuchel, P.W. and Hambly, B.D. and King, G.F.\",\"doi\":\"10.1007/BF00197813\",\"journal\":\"Journal of Biomolecular NMR\",\"number\":\"3\",\"bibtex\":\"@article{\\n title = {Measuring protein self-association using pulsed-field-gradient NMR spectroscopy: Application to myosin light chain 2},\\n type = {article},\\n year = {1995},\\n pages = {321-328},\\n volume = {6},\\n id = {a162714d-04d2-3f71-8080-eeb26343a53a},\\n created = {2023-01-10T01:47:00.198Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:47:00.198Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {At the millimolar concentrations required for structural studies, NMR spectra of the calcium-binding protein myosin light chain 2 (MLC2) showed resonance line widths indicative of extensive self-association. Pulsed-field-gradient (PFG) NMR spectroscopy was used to examine whether MLC2 aggregation could be prevented by the zwitterionic bile salt derivative 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). PFG NMR measurements indicated that CHAPS was capable of preventing MLC2 self-association, but only at concentrations well above the critical micelle concentration of ∼7.5 mM. CHAPS was most effective at a concentration of 22.5 mM, where the apparent molecular mass of MLC2 correponded to a protein monomer plus seven molecules of bound detergent. The resolution and sensitivity of 2D 15N-1H HSQC spectra of MLC2 were markedly improved by the addition of 25 mM CHAPS, consistent with a reduction in aggregation following addition of the detergent. The average amide nitrogen T2 value for MLC2 increased from ∼30 ms in the absence of CHAPS to ∼56 ms in the presence of 25 mM CHAPS. The results of this study lead us to propose that PFG NMR spectroscopy can be used as a facile alternative to conventional techniques such as analytical ultracentrifugation for examining the self-association of biological macromolecules. © 1995 ESCOM Science Publishers B.V.},\\n bibtype = {article},\\n author = {Dingley, A.J. and Mackay, J.P. and Chapman, B.E. and Morris, M.B. and Kuchel, P.W. and Hambly, B.D. and King, G.F.},\\n doi = {10.1007/BF00197813},\\n journal = {Journal of Biomolecular NMR},\\n number = {3}\\n}\",\"author_short\":[\"Dingley,  A.\",\"Mackay,  J.\",\"Chapman,  B.\",\"Morris,  M.\",\"Kuchel,  P.\",\"Hambly,  B.\",\"King,  G.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"dingley-mackay-chapman-morris-kuchel-hambly-king-measuringproteinselfassociationusingpulsedfieldgradientnmrspectroscopyapplicationtomyosinlightchain2-1995\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Functional roles of natural products: The involvement of extended arrays of weak interactions in cooperative binding phenomena\",\"type\":\"article\",\"year\":\"1994\",\"pages\":\"1975-1982\",\"volume\":\"66\",\"id\":\"c10c940c-9f14-3cf0-888e-39c896da615c\",\"created\":\"2023-01-10T01:47:01.128Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:47:01.128Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"A factorisation of the free energy of binding into various “costs” and “benefits” has provided the basis for a semi-quantitation of some weak interactions in solution. It has become clear that the entropie cost of motional restriction on binding (A + B → A.B) increases with increasing exothermicity of the association; this exothermicity must of course reflect not only interactions at the interface between A and B, but also the change in bonding throughout B (if B represents the receptor). We illustrate cooperativity and anti-cooperativity by reference to effects of ligand binding (as models of classical agonists and antagonists) on the dimerisation of vancomycin-group antibiotics. Since dimerization of receptors (promoted by ligand binding) is a common theme in biological signalling, it must presumably have an advantage in natural selection. We suggest that concurrent demands of ligand binding and receptor dimerization may permit a more specific control of those ligand structures which can cause signal transmission. In this way, specificity in biological signalling might be aided. © 1994 IUPAC\",\"bibtype\":\"article\",\"author\":\"Williams, D.H. and Searle, M.S. and Groves, P. and Mackay, J.P. and Westwell, M.S. and Beauregard, D.A. and Cristofaro, M.F.\",\"doi\":\"10.1351/pac199466101975\",\"journal\":\"Pure and Applied Chemistry\",\"number\":\"10-11\",\"bibtex\":\"@article{\\n title = {Functional roles of natural products: The involvement of extended arrays of weak interactions in cooperative binding phenomena},\\n type = {article},\\n year = {1994},\\n pages = {1975-1982},\\n volume = {66},\\n id = {c10c940c-9f14-3cf0-888e-39c896da615c},\\n created = {2023-01-10T01:47:01.128Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:47:01.128Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {A factorisation of the free energy of binding into various “costs” and “benefits” has provided the basis for a semi-quantitation of some weak interactions in solution. It has become clear that the entropie cost of motional restriction on binding (A + B → A.B) increases with increasing exothermicity of the association; this exothermicity must of course reflect not only interactions at the interface between A and B, but also the change in bonding throughout B (if B represents the receptor). We illustrate cooperativity and anti-cooperativity by reference to effects of ligand binding (as models of classical agonists and antagonists) on the dimerisation of vancomycin-group antibiotics. Since dimerization of receptors (promoted by ligand binding) is a common theme in biological signalling, it must presumably have an advantage in natural selection. We suggest that concurrent demands of ligand binding and receptor dimerization may permit a more specific control of those ligand structures which can cause signal transmission. In this way, specificity in biological signalling might be aided. © 1994 IUPAC},\\n bibtype = {article},\\n author = {Williams, D.H. and Searle, M.S. and Groves, P. and Mackay, J.P. and Westwell, M.S. and Beauregard, D.A. and Cristofaro, M.F.},\\n doi = {10.1351/pac199466101975},\\n journal = {Pure and Applied Chemistry},\\n number = {10-11}\\n}\",\"author_short\":[\"Williams,  D.\",\"Searle,  M.\",\"Groves,  P.\",\"Mackay,  J.\",\"Westwell,  M.\",\"Beauregard,  D.\",\"Cristofaro,  M.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"williams-searle-groves-mackay-westwell-beauregard-cristofaro-functionalrolesofnaturalproductstheinvolvementofextendedarraysofweakinteractionsincooperativebindingphenomena-1994\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":0,\"html\":\"\"},{\"title\":\"Dissection of the Contributions toward Dimerization of Glycopeptide Antibiotics\",\"type\":\"article\",\"year\":\"1994\",\"pages\":\"4573-4580\",\"volume\":\"116\",\"id\":\"5793ec9e-c2d9-3eba-9500-f54fc95f52df\",\"created\":\"2023-01-10T01:47:02.045Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:47:02.045Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"A procedure for the determination of association constants in aqueous solution using hydrogen-deuterium exchange has been developed and used to measure the dimerization constant, Kam, for a number of strongly dimerizing glycopeptide antibiotics. These values provide further insight into the thermodynamic contributions of various structural epitopes to the dimerization of these antibiotics. Consideration of ligand binding affinities together with dimerization potentials provides evidence that dimerization is implicated in the physiological mode of action of these antibiotics. © 1994, American Chemical Society. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Gerhard, U. and Beauregard, D.A. and Maplestone, R.A. and Williams, D.H.\",\"doi\":\"10.1021/ja00090a005\",\"journal\":\"Journal of the American Chemical Society\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Dissection of the Contributions toward Dimerization of Glycopeptide Antibiotics},\\n type = {article},\\n year = {1994},\\n pages = {4573-4580},\\n volume = {116},\\n id = {5793ec9e-c2d9-3eba-9500-f54fc95f52df},\\n created = {2023-01-10T01:47:02.045Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:47:02.045Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {A procedure for the determination of association constants in aqueous solution using hydrogen-deuterium exchange has been developed and used to measure the dimerization constant, Kam, for a number of strongly dimerizing glycopeptide antibiotics. These values provide further insight into the thermodynamic contributions of various structural epitopes to the dimerization of these antibiotics. Consideration of ligand binding affinities together with dimerization potentials provides evidence that dimerization is implicated in the physiological mode of action of these antibiotics. © 1994, American Chemical Society. All rights reserved.},\\n bibtype = {article},\\n author = {Mackay, J.P. and Gerhard, U. and Beauregard, D.A. and Maplestone, R.A. and Williams, D.H.},\\n doi = {10.1021/ja00090a005},\\n journal = {Journal of the American Chemical Society},\\n number = {11}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Gerhard,  U.\",\"Beauregard,  D.\",\"Maplestone,  R.\",\"Williams,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-gerhard-beauregard-maplestone-williams-dissectionofthecontributionstowarddimerizationofglycopeptideantibiotics-1994\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":1,\"html\":\"\"},{\"title\":\"Glycopeptide Antibiotic Activity and the Possible Role of Dimerization: A Model for Biological Signaling\",\"type\":\"article\",\"year\":\"1994\",\"pages\":\"4581-4590\",\"volume\":\"116\",\"id\":\"71f945c5-5554-3649-bfe7-e37debf946b9\",\"created\":\"2023-01-10T01:47:02.960Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:47:02.960Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"It is demonstrated that the presence of bacterial cell wall analogues may either enhance or, in the case of ristocetin A, oppose dimerization of glycopeptide antibiotics. These observations may imply that dimerization plays a role in the mode of action of these antibiotics, and a mechanism is proposed to take account of this possibility. The glycopeptide dimers are also found to be formed more exothermically in the presence of cell wall analogues, and the nature of biological signaling events is discussed in this context. It is pointed out that binding enthalpy (rather than simply binding free energy, ΔG) may be an important quantity in signaling events. If this is so, then oligomers may be abundant in signaling processes partly because the extended aggregates they form are able to cooperatively amplify the conformational changes which are incurred on ligand binding, which occur through relatively small changes in free energy but larger opposing changes in enthalpy and entropy. © 1994, American Chemical Society. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Mackay, J.P. and Gerhard, U. and Beauregard, D.A. and Westwell, M.S. and Searle, M.S. and Williams, D.H.\",\"doi\":\"10.1021/ja00090a006\",\"journal\":\"Journal of the American Chemical Society\",\"number\":\"11\",\"bibtex\":\"@article{\\n title = {Glycopeptide Antibiotic Activity and the Possible Role of Dimerization: A Model for Biological Signaling},\\n type = {article},\\n year = {1994},\\n pages = {4581-4590},\\n volume = {116},\\n id = {71f945c5-5554-3649-bfe7-e37debf946b9},\\n created = {2023-01-10T01:47:02.960Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:47:02.960Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {It is demonstrated that the presence of bacterial cell wall analogues may either enhance or, in the case of ristocetin A, oppose dimerization of glycopeptide antibiotics. These observations may imply that dimerization plays a role in the mode of action of these antibiotics, and a mechanism is proposed to take account of this possibility. The glycopeptide dimers are also found to be formed more exothermically in the presence of cell wall analogues, and the nature of biological signaling events is discussed in this context. It is pointed out that binding enthalpy (rather than simply binding free energy, ΔG) may be an important quantity in signaling events. If this is so, then oligomers may be abundant in signaling processes partly because the extended aggregates they form are able to cooperatively amplify the conformational changes which are incurred on ligand binding, which occur through relatively small changes in free energy but larger opposing changes in enthalpy and entropy. © 1994, American Chemical Society. All rights reserved.},\\n bibtype = {article},\\n author = {Mackay, J.P. and Gerhard, U. and Beauregard, D.A. and Westwell, M.S. and Searle, M.S. and Williams, D.H.},\\n doi = {10.1021/ja00090a006},\\n journal = {Journal of the American Chemical Society},\\n number = {11}\\n}\",\"author_short\":[\"Mackay,  J.\",\"Gerhard,  U.\",\"Beauregard,  D.\",\"Westwell,  M.\",\"Searle,  M.\",\"Williams,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"mackay-gerhard-beauregard-westwell-searle-williams-glycopeptideantibioticactivityandthepossibleroleofdimerizationamodelforbiologicalsignaling-1994\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The structure of an asymmetric dimer relevant to the mode of action of the glycopeptide antibiotics\",\"type\":\"article\",\"year\":\"1994\",\"pages\":\"747-754\",\"volume\":\"2\",\"id\":\"19ebea8a-a41d-36ba-9081-a4f636b1f554\",\"created\":\"2023-01-10T01:47:03.895Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:47:03.895Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Background Glycopeptide antibiotics of the vancomycin group are of crucial clinical importance in the treatment of methicillin resistant Staphylococcus aureus (MRSA) - the often lethal 'super-bug ' - characterized by its resistance to a wide range of antibiotics in common use. The antibiotics exert their physiological action by blocking cell wall synthesis through recognition of nascent cell wall mucopeptides terminating in the sequence -D-Ala-D -Ala. Evidence suggests that the antibiotics are able to enhance their biological activity by the formation of homodimers, and this is supported by the observation that dimerization and peptide binding in vitro are cooperative phenomena. The basis of this enhancement is not understood at the molecular level. Results The first detailed structure of a dimeric glycopeptide antibiotic, that of eremomycin, is presented based upon solution NMR data. The overall structure of the dimer complex is asymmetric. The source of this asymmetry - a parallel alignment and mutual interaction of the disaccharides - appears to promote dimerization through specific sugar- sugar recognition. Conclusions A molecular basis for the observed cooperativity of cell wall peptide binding by eremomycin is evident from these studies of the dimer. The carboxylate anion of the cell wall component, which is crucial to binding, forms an amide-mediated ion-pair interaction to the alkylammonium ion of the ring 6 sugar in the other half of the dimer making the structure and positioning of this sugar important in mediating cooperativity. © 1994 Elsevier Science Ltd. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Groves, P. and Searle, M.S. and Mackay, J.P. and Williams, D.H.\",\"doi\":\"10.1016/S0969-2126(94)00075-1\",\"journal\":\"Structure\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {The structure of an asymmetric dimer relevant to the mode of action of the glycopeptide antibiotics},\\n type = {article},\\n year = {1994},\\n pages = {747-754},\\n volume = {2},\\n id = {19ebea8a-a41d-36ba-9081-a4f636b1f554},\\n created = {2023-01-10T01:47:03.895Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:47:03.895Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Background Glycopeptide antibiotics of the vancomycin group are of crucial clinical importance in the treatment of methicillin resistant Staphylococcus aureus (MRSA) - the often lethal 'super-bug ' - characterized by its resistance to a wide range of antibiotics in common use. The antibiotics exert their physiological action by blocking cell wall synthesis through recognition of nascent cell wall mucopeptides terminating in the sequence -D-Ala-D -Ala. Evidence suggests that the antibiotics are able to enhance their biological activity by the formation of homodimers, and this is supported by the observation that dimerization and peptide binding in vitro are cooperative phenomena. The basis of this enhancement is not understood at the molecular level. Results The first detailed structure of a dimeric glycopeptide antibiotic, that of eremomycin, is presented based upon solution NMR data. The overall structure of the dimer complex is asymmetric. The source of this asymmetry - a parallel alignment and mutual interaction of the disaccharides - appears to promote dimerization through specific sugar- sugar recognition. Conclusions A molecular basis for the observed cooperativity of cell wall peptide binding by eremomycin is evident from these studies of the dimer. The carboxylate anion of the cell wall component, which is crucial to binding, forms an amide-mediated ion-pair interaction to the alkylammonium ion of the ring 6 sugar in the other half of the dimer making the structure and positioning of this sugar important in mediating cooperativity. © 1994 Elsevier Science Ltd. All rights reserved.},\\n bibtype = {article},\\n author = {Groves, P. and Searle, M.S. and Mackay, J.P. and Williams, D.H.},\\n doi = {10.1016/S0969-2126(94)00075-1},\\n journal = {Structure},\\n number = {8}\\n}\",\"author_short\":[\"Groves,  P.\",\"Searle,  M.\",\"Mackay,  J.\",\"Williams,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"groves-searle-mackay-williams-thestructureofanasymmetricdimerrelevanttothemodeofactionoftheglycopeptideantibiotics-1994\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"The Role of the Sugar and Chlorine Substituents in the Dimerization of Vancomycin Antibiotics\",\"type\":\"article\",\"year\":\"1993\",\"pages\":\"232-237\",\"volume\":\"115\",\"id\":\"26477609-41c1-3bb1-b211-10fe3f017959\",\"created\":\"2023-01-10T01:47:04.826Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:47:04.826Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"Evidence is presented for the formation of dimers in aqueous solutions of the glycopeptide antibiotics eremomycin, A82846B, vancomycin, and eremomycin-ψ. The dimerization constant Kdim is determined by 1H NMR spectroscopy for the last two compounds and also for the related compound ristocetin-ψ, for which dimerization has previously been reported in mixed solvents. Values of Kdim are obtained for these compounds over a range of temperatures, and thus ΔHdim and ΔSdim are calculated. In addition, a lower limit for Kdim in the case of eremomycin is calculated (105 M−1). This is a remarkably large value, and it may be that dimerization is implicated in antibiotic action. The possibility that natural selection has led to adaptations which promote dimerization (such as the nature and sites of attachment of the sugars and a ring 2 chlorine atom) is discussed. © 1993, American Chemical Society. All rights reserved.\",\"bibtype\":\"article\",\"author\":\"Gerhard, U. and Mackay, J.P. and Maplestone, R.A. and Williams, D.H.\",\"doi\":\"10.1021/ja00054a033\",\"journal\":\"Journal of the American Chemical Society\",\"number\":\"1\",\"bibtex\":\"@article{\\n title = {The Role of the Sugar and Chlorine Substituents in the Dimerization of Vancomycin Antibiotics},\\n type = {article},\\n year = {1993},\\n pages = {232-237},\\n volume = {115},\\n id = {26477609-41c1-3bb1-b211-10fe3f017959},\\n created = {2023-01-10T01:47:04.826Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:47:04.826Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {Evidence is presented for the formation of dimers in aqueous solutions of the glycopeptide antibiotics eremomycin, A82846B, vancomycin, and eremomycin-ψ. The dimerization constant Kdim is determined by 1H NMR spectroscopy for the last two compounds and also for the related compound ristocetin-ψ, for which dimerization has previously been reported in mixed solvents. Values of Kdim are obtained for these compounds over a range of temperatures, and thus ΔHdim and ΔSdim are calculated. In addition, a lower limit for Kdim in the case of eremomycin is calculated (105 M−1). This is a remarkably large value, and it may be that dimerization is implicated in antibiotic action. The possibility that natural selection has led to adaptations which promote dimerization (such as the nature and sites of attachment of the sugars and a ring 2 chlorine atom) is discussed. © 1993, American Chemical Society. All rights reserved.},\\n bibtype = {article},\\n author = {Gerhard, U. and Mackay, J.P. and Maplestone, R.A. and Williams, D.H.},\\n doi = {10.1021/ja00054a033},\\n journal = {Journal of the American Chemical Society},\\n number = {1}\\n}\",\"author_short\":[\"Gerhard,  U.\",\"Mackay,  J.\",\"Maplestone,  R.\",\"Williams,  D.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"gerhard-mackay-maplestone-williams-theroleofthesugarandchlorinesubstituentsinthedimerizationofvancomycinantibiotics-1993\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"downloads\":3,\"html\":\"\"},{\"title\":\"Toward an estimation of binding constants in aqueous solution: Studies of associations of vancomycin group antibiotics\",\"type\":\"article\",\"year\":\"1993\",\"pages\":\"1172-1178\",\"volume\":\"90\",\"id\":\"6e99bb91-e44e-3b3e-a076-022d9dbbba8a\",\"created\":\"2023-01-10T01:47:05.775Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2023-01-10T01:47:05.775Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"private_publication\":false,\"abstract\":\"An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.\",\"bibtype\":\"article\",\"author\":\"Williams, D.H. and Searle, M.S. and Mackay, J.P. and Gerhard, U. and Maplestone, R.A.\",\"doi\":\"10.1073/pnas.90.4.1172\",\"journal\":\"Proceedings of the National Academy of Sciences of the United States of America\",\"number\":\"4\",\"bibtex\":\"@article{\\n title = {Toward an estimation of binding constants in aqueous solution: Studies of associations of vancomycin group antibiotics},\\n type = {article},\\n year = {1993},\\n pages = {1172-1178},\\n volume = {90},\\n id = {6e99bb91-e44e-3b3e-a076-022d9dbbba8a},\\n created = {2023-01-10T01:47:05.775Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2023-01-10T01:47:05.775Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n private_publication = {false},\\n abstract = {An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.},\\n bibtype = {article},\\n author = {Williams, D.H. and Searle, M.S. and Mackay, J.P. and Gerhard, U. and Maplestone, R.A.},\\n doi = {10.1073/pnas.90.4.1172},\\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\\n number = {4}\\n}\",\"author_short\":[\"Williams,  D.\",\"Searle,  M.\",\"Mackay,  J.\",\"Gerhard,  U.\",\"Maplestone,  R.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  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degraders},\\n type = {article},\\n year = {2025},\\n pages = {326-337},\\n volume = {6},\\n id = {30eeaf46-43ed-3d7a-9d00-edc3ea4d8b2e},\\n created = {2025-05-31T09:41:34.856Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2025-05-31T09:41:34.856Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n source_type = {Journal Article},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Jing, X and Mackay, J P and Passioura, T},\\n doi = {10.1039/D4CB00199K},\\n journal = {RSC Chem Biol},\\n number = {3}\\n}\",\"author_short\":[\"Jing,  X.\",\"Mackay,  J., P.\",\"Passioura,  T.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"jing-mackay-passioura-macrocyclicpeptidesasanewclassoftargetedproteindegraders-2025\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  j\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\"}},\"html\":\"\"},{\"title\":\"Rapid Elaboration of Fragments into Leads applied to BRD3-extra terminal domain\",\"type\":\"article\",\"year\":\"2023\",\"pages\":\"5859-5872\",\"volume\":\"66\",\"id\":\"8a395652-12ac-3c77-ab3f-a26469271e65\",\"created\":\"2025-05-31T09:41:35.965Z\",\"file_attached\":false,\"profile_id\":\"a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"group_id\":\"bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\",\"last_modified\":\"2025-05-31T09:41:35.965Z\",\"read\":false,\"starred\":false,\"authored\":false,\"confirmed\":false,\"hidden\":false,\"source_type\":\"Journal Article\",\"private_publication\":false,\"bibtype\":\"article\",\"author\":\"Adams, L A and Wilkinson-White, L E and Gunzburg, M J and Headey, S J and Mohanty, B and Scanlon, M J and Capuano, B and Mackay, J P and Doak, B C\",\"doi\":\"https://doi.org/10.1021/acs.jmedchem.3c00137\",\"journal\":\"Journal of Medicinal Chemistry\",\"number\":\"8\",\"bibtex\":\"@article{\\n title = {Rapid Elaboration of Fragments into Leads applied to BRD3-extra terminal domain},\\n type = {article},\\n year = {2023},\\n pages = {5859-5872},\\n volume = {66},\\n id = {8a395652-12ac-3c77-ab3f-a26469271e65},\\n created = {2025-05-31T09:41:35.965Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2025-05-31T09:41:35.965Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n source_type = {Journal Article},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Adams, L A and Wilkinson-White, L E and Gunzburg, M J and Headey, S J and Mohanty, B and Scanlon, M J and Capuano, B and Mackay, J P and Doak, B C},\\n doi = {https://doi.org/10.1021/acs.jmedchem.3c00137},\\n journal = {Journal of Medicinal Chemistry},\\n number = {8}\\n}\",\"author_short\":[\"Adams,  L., A.\",\"Wilkinson-White,  L., E.\",\"Gunzburg,  M., J.\",\"Headey,  S., J.\",\"Mohanty,  B.\",\"Scanlon,  M., J.\",\"Capuano,  B.\",\"Mackay,  J., P.\",\"Doak,  B., C.\"],\"biburl\":\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386\",\"bibbaseid\":\"adams-wilkinsonwhite-gunzburg-headey-mohanty-scanlon-capuano-mackay-etal-rapidelaborationoffragmentsintoleadsappliedtobrd3extraterminaldomain-2023\",\"role\":\"author\",\"urls\":{},\"metadata\":{\"authorlinks\":{\"mackay,  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K\",\"doi\":\"10.1073/pnas.2219418120\",\"journal\":\"Proceedings of the National Academy of Sciences, USA\",\"number\":\"17\",\"bibtex\":\"@article{\\n title = {Cross-linking mass spectrometry discovers, evaluates, and validates the experimental and predicted structural proteome},\\n type = {article},\\n year = {2023},\\n pages = {e2219418120},\\n volume = {120},\\n id = {9ef702f4-96aa-3165-9804-08012f491b9c},\\n created = {2025-05-31T09:41:37.182Z},\\n file_attached = {false},\\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\\n last_modified = {2025-05-31T09:41:37.182Z},\\n read = {false},\\n starred = {false},\\n authored = {false},\\n confirmed = {false},\\n hidden = {false},\\n source_type = {Journal Article},\\n private_publication = {false},\\n bibtype = {article},\\n author = {Bartolec, T K and Vazquez-Campos, X and Norman, A and Luong, C and Johnson, M and Payne, R J and Wilkins, M R and Mackay, J P and Low, J K 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id=\"bibbase_header\" class=\"dontprint\" style=\"\">\n\n    <ul class=\"nav nav-pills\" style=\"margin: 0px;\">\n\n      <li class=\"dropdown\" id=\"groupby_dropdown\">\n      <a class=\"dropdown-toggle\"\n         data-toggle=\"dropdown\"\n         href=\"#\">\n          Group by\n          <b class=\"caret\"></b>\n      </a>\n      <ul class=\"dropdown-menu\">\n        <li class=\"groupby year\"><a onclick=\"groupby('year')\">\n            Year</a>\n        </li>\n        <li class=\"groupby author\"><a onclick=\"groupby('author_short')\">\n            Author</a>\n        </li>\n        <li class=\"groupby type\"><a onclick=\"groupby('type')\">\n            Type</a>\n        </li>\n        <li class=\"groupby keyword\"><a onclick=\"groupby('keyword')\">\n            Keyword</a>\n        </li>\n        <li class=\"groupby downloads\"><a onclick=\"groupby('downloads')\">\n            Downloads</a>\n        </li>\n      </ul>\n      </li>\n\n\n      <li class=\"dropdown\" 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 type = {inbook},
 year = {2010},
 keywords = {RNA-binding domain,Zinc fingers,protein design,protein interaction domain},
 pages = {479-491},
 volume = {649},
 websites = {http://link.springer.com/10.1007/978-1-60761-753-2_29},
 id = {0f1743b7-0299-3ee3-8152-d528a8b42877},
 created = {2020-12-17T05:29:52.309Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:52.309Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Font2010},
 source_type = {ARTICLE},
 notes = {cited By 21},
 private_publication = {false},
 abstract = {Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.},
 bibtype = {inbook},
 author = {Font, Josep and Mackay, Joel P.},
 doi = {10.1007/978-1-60761-753-2_29},
 chapter = {Beyond DNA: Zinc Finger Domains as RNA-Binding Modules},
 title = {Methods in Molecular Biology}
}

@article{
 title = {1H, 13C and 15N resonance assignments of an N-terminal domain of CHD4},
 type = {article},
 year = {2014},
 keywords = {CHD4,Chromatin remodeling,N-terminal domain,NuRD complex,PAR-binding motif},
 pages = {137-139},
 volume = {8},
 websites = {http://link.springer.com/10.1007/s12104-013-9469-3},
 month = {4},
 day = {17},
 id = {6b604bfa-8cf1-39d7-9959-cdd1ad06e0c1},
 created = {2020-12-17T05:29:52.354Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:52.354Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Silva2014},
 source_type = {ARTICLE},
 notes = {cited By 1},
 private_publication = {false},
 abstract = {Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.},
 bibtype = {article},
 author = {Silva, Ana P. G. and Kwan, Ann H. and Mackay, Joel P.},
 doi = {10.1007/s12104-013-9469-3},
 journal = {Biomolecular NMR Assignments},
 number = {1}
}

@article{
 title = {The Hydrophobin EAS Is Largely Unstructured in Solution and Functions by Forming Amyloid-Like Structures},
 type = {article},
 year = {2001},
 keywords = {Amphipathic monolayer,EAS,Neurospora crassa,Self-assembly,hydrophobin},
 pages = {83-91},
 volume = {9},
 websites = {https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591},
 month = {2},
 id = {298718cc-0cb5-316f-89d4-dead241698cf},
 created = {2020-12-17T05:29:52.461Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:52.461Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Mackay2001},
 source_type = {ARTICLE},
 notes = {cited By 115},
 private_publication = {false},
 abstract = {Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.},
 bibtype = {article},
 author = {Mackay, Joel P and Matthews, Jacqueline M. and Winefield, Robert D. and Mackay, Lindsey G. and Haverkamp, Richard G. and Templeton, Matthew D.},
 doi = {10.1016/S0969-2126(00)00559-1},
 journal = {Structure},
 number = {2}
}

@article{
 title = {Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex},
 type = {article},
 year = {2017},
 keywords = {chromatin remodelling,co-immunoprecipitations,nucleosome remodelling and deacetylase (NuRD) comp,protein structure,protein–protein interactions},
 pages = {4216-4232},
 volume = {284},
 websites = {http://doi.wiley.com/10.1111/febs.14301},
 month = {12},
 id = {fa40e576-8ce6-3e7e-83ec-453c57f0abe1},
 created = {2020-12-17T05:29:52.498Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:52.498Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Torrado2017},
 source_type = {ARTICLE},
 notes = {cited By 4},
 private_publication = {false},
 abstract = {The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.},
 bibtype = {article},
 author = {Torrado, Mario and Low, Jason K. K. and Silva, Ana P. G. and Schmidberger, Jason W. and Sana, Maryam and Sharifi Tabar, Mehdi and Isilak, Musa E. and Winning, Courtney S. and Kwong, Cherry and Bedward, Max J. and Sperlazza, Mary J. and Williams, David C. and Shepherd, Nicholas E. and Mackay, Joel P.},
 doi = {10.1111/febs.14301},
 journal = {The FEBS Journal},
 number = {24}
}

@article{
 title = {Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean},
 type = {article},
 year = {2017},
 keywords = {CLE,CLE40,IAD,arabinosylation,glycopeptide,glycosylation,hormone,legume,root apical meristem,soybean},
 pages = {1347-1355.e7},
 volume = {24},
 websites = {https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124},
 month = {11},
 id = {9da321e9-ae64-34bc-8487-6b2f91bcf82c},
 created = {2020-12-17T05:29:52.604Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:52.604Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Corcilius2017},
 source_type = {ARTICLE},
 notes = {cited By 8},
 private_publication = {false},
 abstract = {Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.},
 bibtype = {article},
 author = {Corcilius, Leo and Hastwell, April H. and Zhang, Mengbai and Williams, James and Mackay, Joel P. and Gresshoff, Peter M. and Ferguson, Brett J. and Payne, Richard J.},
 doi = {10.1016/j.chembiol.2017.08.014},
 journal = {Cell Chemical Biology},
 number = {11}
}

@article{
 title = {The NuRD complex and macrocephaly associated neurodevelopmental disorders},
 type = {article},
 year = {2019},
 keywords = {CHD3,CHD4,GATAD2B,NuRD complex,macrocephaly},
 pages = {548-556},
 volume = {181},
 websites = {http://doi.org/10.1002/ajmg.c.31752},
 month = {12},
 day = {18},
 id = {786626bb-912b-3ca7-aa02-82a98a6afa89},
 created = {2020-12-17T05:29:52.824Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:52.824Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Pierson2019},
 source_type = {article},
 private_publication = {false},
 abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.},
 bibtype = {article},
 author = {Pierson, Tyler Mark and Otero, Maria G. and Grand, Katheryn and Choi, Andrew and Graham, John M. and Young, Juan I. and Mackay, Joel P.},
 doi = {10.1002/ajmg.c.31752},
 journal = {American Journal of Medical Genetics Part C: Seminars in Medical Genetics},
 number = {4}
}

@article{
 title = {Crystallization of a ZRANB2–RNA complex},
 type = {article},
 year = {2008},
 keywords = {RNA-binding proteins,RanBP2-type zinc fingers,Splicing factors},
 pages = {1175-1177},
 volume = {64},
 websites = {http://scripts.iucr.org/cgi-bin/paper?S1744309108036993},
 month = {12},
 day = {1},
 id = {7587f247-d4b3-314f-84e0-30baa59ddf19},
 created = {2020-12-17T05:29:52.864Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:52.864Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Loughlin2008},
 source_type = {ARTICLE},
 notes = {cited By 1},
 private_publication = {false},
 abstract = {ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.},
 bibtype = {article},
 author = {Loughlin, Fionna E. and Lee, Mihwa and Guss, J. Mitchell and Mackay, Joel P.},
 doi = {10.1107/S1744309108036993},
 journal = {Acta Crystallographica Section F Structural Biology and Crystallization Communications},
 number = {12}
}

@article{
 title = {The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain},
 type = {article},
 year = {2016},
 keywords = {C17orf64,DNA-binding domain,helical bundle,nucleosomes,transcription},
 pages = {4298-4314},
 volume = {428},
 websites = {https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485},
 month = {10},
 id = {851b1fbc-10b7-3289-b546-77e83ffb0c9d},
 created = {2020-12-17T05:29:53.207Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:53.207Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Mohanty2016},
 source_type = {ARTICLE},
 notes = {cited By 5},
 private_publication = {false},
 abstract = {The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.},
 bibtype = {article},
 author = {Mohanty, Biswaranjan and Helder, Stephanie and Silva, Ana P.G. and Mackay, Joel P. and Ryan, Daniel P.},
 doi = {10.1016/j.jmb.2016.08.028},
 journal = {Journal of Molecular Biology},
 number = {21}
}

@article{
 title = {GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder},
 type = {article},
 year = {2020},
 keywords = {GATAD2B,NuRD complex,apraxia of speech,chromati},
 pages = {878-888},
 volume = {22},
 websites = {http://doi.org/10.1038/s41436-019-0747-z},
 month = {5},
 day = {17},
 id = {6308af4d-95ba-3986-b5f1-b8362afa6d89},
 created = {2020-12-17T05:29:53.237Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:53.237Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Shieh2020},
 source_type = {article},
 private_publication = {false},
 abstract = {Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.},
 bibtype = {article},
 author = {Shieh, Christine and Jones, Natasha and Vanle, Brigitte and Au, Margaret and Huang, Alden Y. and Silva, Ana P. G. and Lee, Hane and Douine, Emilie D. and Otero, Maria G. and Choi, Andrew and Grand, Katheryn and Taff, Ingrid P. and Delgado, Mauricio R. and Hajianpour, M. J. and Seeley, Andrea and Rohena, Luis and Vernon, Hilary and Gripp, Karen W. and Vergano, Samantha A. and Mahida, Sonal and Naidu, Sakkubai and Sousa, Ana Berta and Wain, Karen E. and Challman, Thomas D. and Beek, Geoffrey and Basel, Donald and Ranells, Judith and Smith, Rosemarie and Yusupov, Roman and Freckmann, Mary-Louise and Ohden, Lisa and Davis-Keppen, Laura and Chitayat, David and Dowling, James J. and Finkel, Richard and Dauber, Andrew and Spillmann, Rebecca and Pena, Loren D. M. and Metcalfe, Kay and Splitt, Miranda and Lachlan, Katherine and McKee, Shane A. and Hurst, Jane and Fitzpatrick, David R. and Morton, Jenny E. V. and Cox, Helen and Venkateswaran, Sunita and Young, Juan I. and Marsh, Eric D. and Nelson, Stanley F. and Martinez, Julian A. and Graham, John M. and Kini, Usha and Mackay, Joel P. and Pierson, Tyler Mark},
 doi = {10.1038/s41436-019-0747-z},
 journal = {Genetics in Medicine},
 number = {5}
}

@article{
 title = {Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P},
 type = {article},
 year = {2018},
 keywords = {NLR [nucleotide-binding and oligomerization domain,crystal structure,effector-triggered immunity,flax rust (Melampsora lini) effector,nuclear localization,nucleotide-binding/leucine-rich repeat receptor],plant disease resistance,zinc finger},
 pages = {1196-1209},
 volume = {19},
 websites = {http://doi.wiley.com/10.1111/mpp.12597},
 month = {5},
 id = {67c03f3e-7a41-34e7-89cf-55716f13ac32},
 created = {2020-12-17T05:29:53.248Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:53.248Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Zhang2018},
 source_type = {ARTICLE},
 notes = {cited By 2},
 private_publication = {false},
 abstract = {The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.},
 bibtype = {article},
 author = {Zhang, Xiaoxiao and Farah, Nadya and Rolston, Laura and Ericsson, Daniel J. and Catanzariti, Ann-Maree and Bernoux, Maud and Ve, Thomas and Bendak, Katerina and Chen, Chunhong and Mackay, Joel P. and Lawrence, Gregory J. and Hardham, Adrienne and Ellis, Jeffrey G. and Williams, Simon J. and Dodds, Peter N. and Jones, David A. and Kobe, Bostjan},
 doi = {10.1111/mpp.12597},
 journal = {Molecular Plant Pathology},
 number = {5}
}

@article{
 title = {Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/Friend of GATA interaction},
 type = {article},
 year = {2005},
 keywords = {Factor,Gene expression,Transcription},
 pages = {583-588},
 volume = {102},
 websites = {http://www.ncbi.nlm.nih.gov/pubmed/15644435},
 month = {1},
 day = {18},
 id = {4fa93b4d-9743-397c-8bc8-7dec216c8aa7},
 created = {2020-12-17T05:29:53.577Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:53.577Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Liew2005},
 source_type = {ARTICLE},
 notes = {cited By 65},
 private_publication = {false},
 abstract = {GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.},
 bibtype = {article},
 author = {Liew, Chu Kong and Simpson, Raina J Y and Kwan, Ann H Y and Crofts, Linda A. and Loughlin, Fionna E. and Matthews, Jacqueline M. and Crossley, Merlin and Mackay, Joel P.},
 doi = {10.1073/pnas.0407511102},
 journal = {Proceedings of the National Academy of Sciences},
 number = {3}
}

@article{
 title = {DNA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II},
 type = {article},
 year = {1997},
 keywords = {CD,DNA chemical footprinting,Fotemiistine,Intercalation,Linear dichroism,RNA polymerase II,SPXX peptides,YSPTSPSY,β-tum},
 pages = {387-398},
 volume = {42},
 websites = {https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M},
 month = {10},
 day = {5},
 id = {987f67ee-49d8-3803-8df7-9251cdbaf6c9},
 created = {2020-12-17T05:29:53.650Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:53.650Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Harding1997},
 source_type = {ARTICLE},
 notes = {cited By 5},
 private_publication = {false},
 abstract = {The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley & Sons, Inc.},
 bibtype = {article},
 author = {Harding, Margaret M. and Krippner, Guy Y. and Shelton, Cathryn J. and Rodger, Alison and Sanders, Karen J. and Mackay, Joel P. and Prakash, Arungundrum S.},
 doi = {10.1002/(SICI)1097-0282(19971005)42:4<387::AID-BIP2>3.0.CO;2-M},
 journal = {Biopolymers},
 number = {4}
}

@article{
 title = {Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry},
 type = {article},
 year = {2012},
 keywords = {Mycobacteria,PIN-domain,RNA interferase,RNase,Toxin-antitoxin,VapC},
 pages = {1267-1278},
 volume = {18},
 websites = {http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111},
 month = {6},
 day = {1},
 id = {52a8f59e-942d-361e-8a56-5a3ea03f061e},
 created = {2020-12-17T05:29:53.840Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:53.840Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {McKenzie2012},
 source_type = {ARTICLE},
 notes = {cited By 31},
 private_publication = {false},
 abstract = {The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.},
 bibtype = {article},
 author = {McKenzie, J. L. and Duyvestyn, Johanna M. and Smith, Tony and Bendak, Katerina and MacKay, J. and Cursons, R. and Cook, Gregory M. and Arcus, Vickery L.},
 doi = {10.1261/rna.031229.111},
 journal = {RNA},
 number = {6}
}

@article{
 title = {Interdomain orientation of cardiac Troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state},
 type = {article},
 year = {2012},
 keywords = {Cardiac troponin C,Ensemble states,Paramagnetic relaxation enhancement,Site-directed spin labeling,Solution NMR},
 pages = {1376-1387},
 volume = {21},
 websites = {http://doi.wiley.com/10.1002/pro.2124},
 month = {9},
 id = {136eec2a-03b6-3cd9-bce5-f2eed2bdbc99},
 created = {2020-12-17T05:29:54.085Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:54.085Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Cordina2012},
 source_type = {ARTICLE},
 notes = {cited By 9},
 private_publication = {false},
 abstract = {Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.},
 bibtype = {article},
 author = {Cordina, Nicole M. and Liew, Chu Kong and Gell, David A. and Fajer, Piotr G. and Mackay, Joel P. and Brown, Louise J.},
 doi = {10.1002/pro.2124},
 journal = {Protein Science},
 number = {9}
}

@inbook{
 type = {inbook},
 year = {2010},
 keywords = {Coupled folding and binding,Hotspots,Post-translational modifications,Protein interfaces,Protein recognition},
 pages = {505-532},
 volume = {2},
 websites = {http://doi.wiley.com/10.1002/9783527631780.ch12},
 month = {12},
 publisher = {Wiley-VCH Verlag GmbH & Co. KGaA},
 day = {28},
 city = {Weinheim, Germany},
 id = {31798292-fed8-39f9-8349-947a9649059a},
 created = {2020-12-17T05:29:54.383Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:54.383Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Mansfield2010},
 source_type = {BOOK},
 notes = {cited By 0},
 private_publication = {false},
 bibtype = {inbook},
 author = {Mansfield, Robyn E. and Cross, Arwen J. and Matthews, Jacqueline M. and Mackay, Joel P.},
 doi = {10.1002/9783527631780.ch12},
 chapter = {Protein Recognition},
 title = {Amino Acids, Peptides and Proteins in Organic Chemistry}
}

@article{
 title = {Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications},
 type = {article},
 year = {2009},
 keywords = {Chromodomain helicase DNA-binding protein 4 (CHD4),Histone,Methylation,Plant homeodomain (PHD)},
 pages = {179-187},
 volume = {423},
 websites = {https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is},
 month = {10},
 day = {15},
 id = {4c2da44b-b1f6-3dcb-a479-0d561aabb87c},
 created = {2020-12-17T05:29:54.740Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:54.740Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Musselman2009},
 source_type = {ARTICLE},
 notes = {cited By 73},
 private_publication = {false},
 abstract = {CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation or methylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2–H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin.},
 bibtype = {article},
 author = {Musselman, Catherine A. and Mansfield, Robyn E. and Garske, Adam L. and Davrazou, Foteini and Kwan, Ann H. and Oliver, Samuel S. and O'Leary, Heather and Denu, John M. and Mackay, Joel P. and Kutateladze, Tatiana G.},
 doi = {10.1042/BJ20090870},
 journal = {Biochemical Journal},
 number = {2}
}

@article{
 title = {The structural analysis of proteinâ&#128;&#147;protein interactions by NMR spectroscopy},
 type = {article},
 year = {2009},
 keywords = {NMR spectroscopy,Protein-protein interactions,Structural biology,Technology},
 pages = {5224-5232},
 volume = {9},
 websites = {http://doi.wiley.com/10.1002/pmic.200900303},
 month = {12},
 id = {fc390463-14cb-327f-93bf-fbec6b3cd8fd},
 created = {2020-12-17T05:29:55.049Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:55.049Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {OConnell2009},
 source_type = {ARTICLE},
 notes = {cited By 44},
 private_publication = {false},
 abstract = {A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.},
 bibtype = {article},
 author = {O'Connell, Mitchell R. and Gamsjaeger, Roland and Mackay, Joel P.},
 doi = {10.1002/pmic.200900303},
 journal = {PROTEOMICS},
 number = {23}
}

@article{
 title = {Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana},
 type = {article},
 year = {2018},
 keywords = {DNA binding affinity,DOF zinc finger domain,Gel retardation assay,Microscale thermophoresis},
 pages = {167-176},
 volume = {8},
 websites = {http://bi.tbzmed.ac.ir/Abstract/bi-17551},
 month = {1},
 day = {28},
 id = {dd4675dd-58e2-3545-8030-ebe4c5b425a8},
 created = {2020-12-17T05:29:55.192Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:55.192Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {MoghaddasSani2018},
 source_type = {ARTICLE},
 notes = {cited By 1},
 private_publication = {false},
 abstract = {Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.},
 bibtype = {article},
 author = {Moghaddas Sani, Hakimeh and Hamzeh-Mivehroud, Maryam and Silva, Ana P. and Walshe, James L. and Mohammadi, S. Abolghasem and Rahbar-Shahrouziasl, Mahdyieh and Abbasi, Milad and Jamshidi, Omid and Low, Jason K.K. and Dastmalchi, Siavoush and Mackay, Joel P.},
 doi = {10.15171/bi.2018.19},
 journal = {BioImpacts},
 number = {3}
}

@article{
 title = {Presence of transient helical segments in the galanin-like peptide evident from 1H NMR, circular dichroism, and prediction studies},
 type = {article},
 year = {2004},
 keywords = {Circular dichroism,Galanin-like peptide,NMR spectroscopy,Nascent helix,Secondary structure prediction},
 pages = {261-271},
 volume = {146},
 websites = {http://www.ncbi.nlm.nih.gov/pubmed/15099568},
 month = {6},
 id = {ccd45448-027a-3ba1-94ee-88d2de266d92},
 created = {2020-12-17T05:29:55.417Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:55.417Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Dastmalchi2004},
 source_type = {ARTICLE},
 notes = {cited By 7},
 private_publication = {false},
 abstract = {Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods.},
 bibtype = {article},
 author = {Dastmalchi, Siavoush and Church, W.Bret and Morris, Michael B. and Iismaa, Tiina P. and Mackay, Joel P.},
 doi = {10.1016/j.jsb.2004.01.004},
 journal = {Journal of Structural Biology},
 number = {3}
}

@article{
 title = {Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1},
 type = {article},
 year = {2011},
 keywords = {Haematopoiesis,Protein-DNA interactions,Protein-protein interactions,Transcription factor complex},
 pages = {14443-14448},
 volume = {108},
 websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108},
 month = {8},
 day = {30},
 id = {a786b1a3-bd30-3c66-8aff-7ffb5cbbb0da},
 created = {2020-12-17T05:29:55.512Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:55.512Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Wilkinson-White2011},
 source_type = {ARTICLE},
 notes = {cited By 24},
 private_publication = {false},
 abstract = {The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (<50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.},
 bibtype = {article},
 author = {Wilkinson-White, Lorna and Gamsjaeger, Roland and Dastmalchi, Siavoush and Wienert, Beeke and Stokes, Philippa H. and Crossley, Merlin and Mackay, Joel P. and Matthews, Jacqueline M.},
 doi = {10.1073/pnas.1105898108},
 journal = {Proceedings of the National Academy of Sciences},
 number = {35}
}

@article{
 title = {Peppy: A virtual reality environment for exploring the principles of polypeptide structure},
 type = {article},
 year = {2020},
 keywords = {polypeptide,protein,secondary structure,teaching,undergraduate,virtual reality},
 pages = {157-168},
 volume = {29},
 websites = {http://www.ncbi.nlm.nih.gov/pubmed/31622516},
 month = {1},
 day = {11},
 id = {fe01e53b-d05d-3939-8395-e8da99c6004e},
 created = {2020-12-17T05:29:55.556Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:55.556Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Doak2020},
 source_type = {article},
 private_publication = {false},
 abstract = {A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.},
 bibtype = {article},
 author = {Doak, David G. and Denyer, Gareth S. and Gerrard, Juliet A. and Mackay, Joel P. and Allison, Jane R.},
 doi = {10.1002/pro.3752},
 journal = {Protein Science},
 number = {1}
}

@article{
 title = {Engineering Specificity Changes on a RanBP2 Zinc Finger that Binds Single-Stranded RNA},
 type = {article},
 year = {2014},
 keywords = {RNA binding,combinatorial chemistry,phage display,protein design,zinc fingers},
 pages = {7848-7852},
 volume = {53},
 websites = {http://doi.wiley.com/10.1002/anie.201402980},
 month = {7},
 day = {21},
 id = {5c546e1a-664d-3c49-b780-ca530f662b55},
 created = {2020-12-17T05:29:55.706Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:55.706Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Vandevenne2014},
 source_type = {ARTICLE},
 notes = {cited By 3},
 private_publication = {false},
 abstract = {The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.},
 bibtype = {article},
 author = {Vandevenne, Marylène and O'Connell, Mitchell R. and Helder, Stephanie and Shepherd, Nicholas E. and Matthews, Jacqueline M. and Kwan, Ann H. and Segal, David J. and Mackay, Joel P.},
 doi = {10.1002/anie.201402980},
 journal = {Angewandte Chemie International Edition},
 number = {30}
}

@article{
 title = {The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein},
 type = {article},
 year = {2016},
 keywords = {RNA binding,RNA polymerase,SELEX,Spt4/5,transcription elongation,transcription elongation factor},
 pages = {1710-1721},
 volume = {25},
 websites = {http://doi.wiley.com/10.1002/pro.2976},
 month = {9},
 id = {cb1f5362-a6b4-3c3e-a896-fd523c0b1404},
 created = {2020-12-17T05:29:56.143Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:56.143Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Blythe2016},
 source_type = {ARTICLE},
 notes = {cited By 2},
 private_publication = {false},
 abstract = {The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.},
 bibtype = {article},
 author = {Blythe, Amanda J. and Yazar-Klosinski, Berra and Webster, Michael W. and Chen, Eefei and Vandevenne, Marylène and Bendak, Katerina and Mackay, Joel P. and Hartzog, Grant A. and Vrielink, Alice},
 doi = {10.1002/pro.2976},
 journal = {Protein Science},
 number = {9}
}

@article{
 title = {Backbone and sidechain 1H, 13C and 15N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans},
 type = {article},
 year = {2012},
 keywords = {DewA,Functional amyloid,Hydrophobin,NMR assignment},
 pages = {83-86},
 volume = {6},
 websites = {http://link.springer.com/10.1007/s12104-011-9330-5},
 month = {4},
 day = {4},
 id = {852baf30-6704-30e4-8fdc-8cbe794bb64a},
 created = {2020-12-17T05:29:56.237Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:56.237Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Morris2012},
 source_type = {ARTICLE},
 notes = {cited By 4},
 private_publication = {false},
 abstract = {Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.},
 bibtype = {article},
 author = {Morris, Vanessa K. and Kwan, Ann H. and Mackay, Joel P. and Sunde, Margaret},
 doi = {10.1007/s12104-011-9330-5},
 journal = {Biomolecular NMR Assignments},
 number = {1}
}

@article{
 title = {Ikaros: a key regulator of haematopoiesis},
 type = {article},
 year = {2002},
 keywords = {Haematopoiesis,Ikaros,Pericentromeric-heterochromatin,Transcription},
 pages = {1304-1307},
 volume = {34},
 websites = {http://www.ncbi.nlm.nih.gov/pubmed/12127581},
 month = {10},
 id = {5eef5524-c104-30d5-8fd5-93d7f32bc139},
 created = {2020-12-17T05:29:56.237Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:56.237Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Westman2002},
 source_type = {ARTICLE},
 notes = {cited By 21},
 private_publication = {false},
 abstract = {Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function--unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription.},
 bibtype = {article},
 author = {Westman, Belinda J. and Mackay, Joel P. and Gell, David},
 doi = {10.1016/S1357-2725(02)00070-5},
 journal = {The International Journal of Biochemistry & Cell Biology},
 number = {10}
}

@article{
 title = {Solution structure of a tethered Lmo2 LIM2 /Ldb1 LID complex},
 type = {article},
 year = {2012},
 keywords = {Ldb1,Lmo2,Modular binding,NMR structure},
 pages = {1768-1774},
 volume = {21},
 websites = {http://doi.wiley.com/10.1002/pro.2153},
 month = {11},
 id = {574f81df-0620-3c4a-af49-474d48a6b829},
 created = {2020-12-17T05:29:56.294Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:56.294Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Dastmalchi2012},
 source_type = {ARTICLE},
 notes = {cited By 7},
 private_publication = {false},
 abstract = {LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.},
 bibtype = {article},
 author = {Dastmalchi, Siavoush and Wilkinson-White, Lorna and Kwan, Ann H. and Gamsjaeger, Roland and Mackay, Joel P. and Matthews, Jacqueline M.},
 doi = {10.1002/pro.2153},
 journal = {Protein Science},
 number = {11}
}

@article{
 title = {A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells},
 type = {article},
 year = {2015},
 keywords = {Affinity purification,Friend of GATA1 (FOG1),Multi-protein complex,Nucleosome remodeling and deacetylase complex pept},
 pages = {960-965},
 volume = {23},
 websites = {https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395},
 month = {3},
 id = {c7a33bb8-06cd-300f-9f85-c79fbd926928},
 created = {2020-12-17T05:29:56.497Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:56.497Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Saathoff2015},
 source_type = {ARTICLE},
 notes = {cited By 5},
 private_publication = {false},
 abstract = {We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).},
 bibtype = {article},
 author = {Saathoff, Hinnerk and Brofelth, Mattias and Trinh, Anne and Parker, Benjamin L. and Ryan, Daniel P. and Low, Jason K.K. and Webb, Sarah R. and Silva, Ana P.G. and Mackay, Joel P. and Shepherd, Nicholas E.},
 doi = {10.1016/j.bmc.2015.01.023},
 journal = {Bioorganic & Medicinal Chemistry},
 number = {5}
}

@article{
 title = {Grb7-SH2 domain dimerisation is affected by a single point mutation.},
 type = {article},
 year = {2005},
 keywords = {Analytical ultracentrifugation,Growth factor receptor bound protein 7,Protein engineering,Size-exclusion chromatography,Src homology 2 domain},
 pages = {454-60},
 volume = {34},
 websites = {http://www.ncbi.nlm.nih.gov/pubmed/15841400},
 month = {7},
 day = {20},
 id = {042bb7c3-136a-3d03-a241-4b3ebfa7ee45},
 created = {2020-12-17T05:29:56.590Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:56.590Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Porter2005},
 source_type = {ARTICLE},
 notes = {cited By 20},
 private_publication = {false},
 abstract = {Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 muM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation.},
 bibtype = {article},
 author = {Porter, Corrine J. and Wilce, Matthew C J and Mackay, Joel P. and Leedman, Peter and Wilce, Jackie A.},
 doi = {10.1007/s00249-005-0480-1},
 journal = {European biophysics journal : EBJ},
 number = {5}
}

@article{
 title = {1H, 13C and 15N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis},
 type = {article},
 year = {2010},
 keywords = {Bacillus subtilis,Bacterial signal transduction,Histidine kinase inhibition,KipI},
 pages = {167-169},
 volume = {4},
 websites = {http://link.springer.com/10.1007/s12104-010-9237-6},
 month = {10},
 day = {4},
 id = {48c78b62-9179-35cb-ae9d-83d16d7e9f3b},
 created = {2020-12-17T05:29:56.688Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:56.688Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Hynson2010},
 source_type = {ARTICLE},
 notes = {cited By 2},
 private_publication = {false},
 abstract = {KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.},
 bibtype = {article},
 author = {Hynson, Robert M. G. and Kwan, Ann H. and Jacques, David A. and Mackay, Joel P. and Trewhella, Jill},
 doi = {10.1007/s12104-010-9237-6},
 journal = {Biomolecular NMR Assignments},
 number = {2}
}

@article{
 title = {The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG.},
 type = {article},
 year = {1999},
 keywords = {GATA-1,Protein-protein interactions,Solution structure,Zinc finger},
 pages = {249-62},
 volume = {13},
 websites = {http://www.ncbi.nlm.nih.gov/pubmed/10212985},
 month = {3},
 id = {1bc4df8b-a093-3a22-ba88-996d8029dfd5},
 created = {2020-12-17T05:29:56.853Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:56.853Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Kowalski1999},
 source_type = {ARTICLE},
 notes = {cited By 48},
 private_publication = {false},
 abstract = {Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted beta-hairpins and a single alpha-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA-1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.},
 bibtype = {article},
 author = {Kowalski, K. and Czolij, R. and King, G. F. and Crossley, M. and Mackay, J. P.},
 doi = {https://doi.org/10.1023/a:1008309602929},
 journal = {Journal of Biomolecular NMR},
 number = {3}
}

@article{
 title = {Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes},
 type = {article},
 year = {2011},
 keywords = {Gene regulation,Hematopoiesis,Posttranslational modifications},
 pages = {E159-E168},
 volume = {108},
 websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108},
 month = {5},
 day = {31},
 id = {f5506c6c-a1c6-37df-8e6d-aee9893f4e46},
 created = {2020-12-17T05:29:57.064Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:57.064Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Lamonica2011},
 source_type = {ARTICLE},
 notes = {cited By 98},
 private_publication = {false},
 abstract = {Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 "reads" acetyl marks on nuclear factors to promote their stable association with chromatin.},
 bibtype = {article},
 author = {Lamonica, Janine M. and Deng, Wulan and Kadauke, Stephan and Campbell, Amy E. and Gamsjaeger, Roland and Wang, Hongxin and Cheng, Yong and Billin, Andrew N. and Hardison, Ross C. and Mackay, Joel P. and Blobel, Gerd A.},
 doi = {10.1073/pnas.1102140108},
 journal = {Proceedings of the National Academy of Sciences},
 number = {22}
}

@article{
 title = {Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1},
 type = {article},
 year = {2014},
 keywords = {Breast cancer,DEAF-1,Embryonic development,LMO4,Transcriptional complex},
 pages = {141-144},
 volume = {8},
 websites = {http://link.springer.com/10.1007/s12104-013-9470-x},
 month = {4},
 day = {16},
 id = {70636852-15f1-3af2-95ff-10b3cc12cad6},
 created = {2020-12-17T05:29:57.300Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:57.300Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Joseph2014a},
 source_type = {ARTICLE},
 notes = {cited By 2},
 private_publication = {false},
 abstract = {The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.},
 bibtype = {article},
 author = {Joseph, Soumya and Kwan, Ann H. Y. and Mackay, Joel P. and Cubeddu, Liza and Matthews, Jacqueline M.},
 doi = {10.1007/s12104-013-9470-x},
 journal = {Biomolecular NMR Assignments},
 number = {1}
}

@article{
 title = {Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding},
 type = {article},
 year = {2018},
 keywords = {P2RY12,blood platelet disorder,human,inherited,platelet dysfunction},
 pages = {44-53},
 volume = {16},
 websites = {http://doi.wiley.com/10.1111/jth.13900},
 month = {1},
 id = {6ba6e7ee-21d0-3efa-a162-8e61a57a2e8e},
 created = {2020-12-17T05:29:57.411Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:57.411Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Mundell2018},
 source_type = {ARTICLE},
 notes = {cited By 1},
 private_publication = {false},
 abstract = {Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.},
 bibtype = {article},
 author = {Mundell, S. J. and Rabbolini, D. and Gabrielli, S. and Chen, Q. and Aungraheeta, R. and Hutchinson, J. L. and Kilo, T. and Mackay, J. and Ward, C. M. and Stevenson, W. and Morel-Kopp, M.-C.},
 doi = {10.1111/jth.13900},
 journal = {Journal of Thrombosis and Haemostasis},
 number = {1}
}

@article{
 title = {Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR},
 type = {article},
 year = {1997},
 keywords = {Macromolecules,Pulsed-field-gradient NMR,Self-association,Solvent suppression,Translational diffusion coefficient},
 pages = {1-8},
 volume = {10},
 websites = {https://link.springer.com/article/10.1023/A:1018339526108},
 id = {c4559b9b-27f6-3507-a99a-884b885fa4fb},
 created = {2020-12-17T05:29:57.841Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:57.841Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Dingley1997},
 source_type = {ARTICLE},
 notes = {cited By 24},
 private_publication = {false},
 abstract = {We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.},
 bibtype = {article},
 author = {Dingley, Andrew J. and Mackay, Joel P. and Shaw, Graeme L. and Hambly, Brett D. and King, Glenn F.},
 doi = {https://doi.org/10.1023/A:1018339526108},
 journal = {Journal of Biomolecular NMR},
 number = {1}
}

@article{
 title = {The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G},
 type = {article},
 year = {2019},
 keywords = {IPR020489,NMR,PF13989,RNA recognition motif,aminoglycoside,elongation factor},
 pages = {699-705},
 volume = {87},
 websites = {http://doi.org/10.1002/prot.25687},
 month = {8},
 day = {22},
 id = {7fb760d5-9bf0-3ae6-b357-d8046463887d},
 created = {2020-12-17T05:29:57.892Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:57.892Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Mohanty2019},
 source_type = {article},
 private_publication = {false},
 abstract = {InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.},
 bibtype = {article},
 author = {Mohanty, Biswaranjan and Hanson‐Manful, Paulina and Finn, Thomas J. and Chambers, Cecilia R. and McKellar, James L. O. and Macindoe, Ingrid and Helder, Stephanie and Setiyaputra, Surya and Zhong, Yichen and Mackay, Joel P. and Patrick, Wayne M.},
 doi = {10.1002/prot.25687},
 journal = {Proteins: Structure, Function, and Bioinformatics},
 number = {8}
}

@article{
 title = {Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5},
 type = {article},
 year = {2014},
 keywords = {Expression and purification,KOW domain,Spt4/5,Ubiquitin tag,pHUE},
 pages = {54-60},
 volume = {100},
 websites = {https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989},
 month = {8},
 id = {c8758ecd-f201-3acf-9f44-56e1000a4f6a},
 created = {2020-12-17T05:29:57.964Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:57.964Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Blythe2014},
 source_type = {ARTICLE},
 notes = {cited By 1},
 private_publication = {false},
 abstract = {Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.},
 bibtype = {article},
 author = {Blythe, Amanda and Gunasekara, Sanjika and Walshe, James and Mackay, Joel P. and Hartzog, Grant A. and Vrielink, Alice},
 doi = {10.1016/j.pep.2014.05.005},
 journal = {Protein Expression and Purification}
}

@article{
 title = {Toward an estimation of binding constants in aqueous solution: studies of associations of vancomycin group antibiotics.},
 type = {article},
 year = {1993},
 keywords = {Amide-amide hydrogen bond strengths,Enthalpic barriers,Enthalpy,Entropy compensation,Hydrophobic effect,Rotor restrictions},
 pages = {1172-1178},
 volume = {90},
 websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172},
 month = {2},
 day = {15},
 id = {5ef1cb1b-6f04-35c7-955d-7c5ca2d128fe},
 created = {2020-12-17T05:29:58.125Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:58.125Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Williams1993},
 source_type = {ARTICLE},
 notes = {cited By 160},
 private_publication = {false},
 abstract = {An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.},
 bibtype = {article},
 author = {Williams, Dudley H. and Searle, Mark S. and Mackay, Joel P. and Gerhard, Ute and Maplestone, Rachel A.},
 doi = {10.1073/pnas.90.4.1172},
 journal = {Proceedings of the National Academy of Sciences},
 number = {4}
}

@inbook{
 type = {inbook},
 year = {2018},
 keywords = {Chemical exchange,Chemical shift perturbation,Cross-saturation,Dark states,Macromolecular NMR spectroscopy,Methyl-TROSY,Protein complexes,Protein-protein interactions},
 pages = {2099-2132},
 websites = {http://link.springer.com/10.1007/978-3-319-28388-3_121},
 publisher = {Springer International Publishing},
 city = {Cham},
 id = {49e6d065-5bfa-3734-ac4c-302065de5fd0},
 created = {2020-12-17T05:29:58.281Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:58.281Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Gell2018},
 source_type = {article},
 private_publication = {false},
 abstract = {Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.},
 bibtype = {inbook},
 author = {Gell, David A. and Kwan, Ann H. and Mackay, Joel P.},
 doi = {10.1007/978-3-319-28388-3_121},
 chapter = {NMR Spectroscopy in the Analysis of Protein-Protein Interactions},
 title = {Modern Magnetic Resonance}
}

@article{
 title = {Solution Structures of Two CCHC Zinc Fingers from the FOG Family Protein U-Shaped that Mediate Protein–Protein Interactions},
 type = {article},
 year = {2000},
 keywords = {GATA-1,Structure,Transcription,U-shaped,Zinc fingers},
 pages = {1157-1166},
 volume = {8},
 websites = {https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X},
 month = {11},
 id = {ee4bb2bf-b119-390f-a5c0-747836832737},
 created = {2020-12-17T05:29:58.497Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:58.497Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Liew2000},
 source_type = {ARTICLE},
 notes = {cited By 34},
 private_publication = {false},
 abstract = {Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.},
 bibtype = {article},
 author = {Liew, Chu Kong and Kowalski, Kasper and Fox, Archa H. and Newton, Anthea and Sharpe, Belinda K. and Crossley, Merlin and Mackay, Joel P.},
 doi = {10.1016/S0969-2126(00)00527-X},
 journal = {Structure},
 number = {11}
}

@article{
 title = {Structural basis for rodlet assembly in fungal hydrophobins},
 type = {article},
 year = {2006},
 keywords = {Amyloid,NMR,Polymer},
 pages = {3621-3626},
 volume = {103},
 websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103},
 month = {3},
 day = {7},
 id = {5c3390e3-fea1-3491-b3cb-82c763c2d1b5},
 created = {2020-12-17T05:29:58.566Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:58.566Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Kwan2006},
 source_type = {ARTICLE},
 notes = {cited By 150},
 private_publication = {false},
 abstract = {Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.},
 bibtype = {article},
 author = {Kwan, A. H. Y. and Winefield, R. D. and Sunde, M. and Matthews, J. M. and Haverkamp, R. G. and Templeton, M. D. and Mackay, J. P.},
 doi = {10.1073/pnas.0505704103},
 journal = {Proceedings of the National Academy of Sciences},
 number = {10}
}

@article{
 title = {A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP},
 type = {article},
 year = {2006},
 keywords = {NMR,haem,haem-binding protein,p22HBP,protein structure},
 pages = {287-297},
 volume = {362},
 websites = {https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679},
 month = {9},
 id = {6eee0455-9c97-35b4-ad59-54f7f318abd4},
 created = {2020-12-17T05:29:58.858Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:58.858Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Gell2006},
 source_type = {ARTICLE},
 notes = {cited By 6},
 private_publication = {false},
 abstract = {The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Gell, David A. and Westman, Belinda J. and Gorman, Daniel and Liew, ChuKong and Welch, John J. and Weiss, Mitchell J. and Mackay, Joel P.},
 doi = {10.1016/j.jmb.2006.07.010},
 journal = {Journal of Molecular Biology},
 number = {2}
}

@article{
 title = {GATA-1: One protein, many partners},
 type = {article},
 year = {2006},
 keywords = {Factor,Finger,Hematopoiesis,Transcription,Zinc},
 pages = {6-11},
 volume = {38},
 websites = {https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074},
 month = {1},
 id = {ef48217b-ded1-36e7-b8a7-8426ac4c459e},
 created = {2020-12-17T05:29:58.983Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:58.983Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Lowry2006},
 source_type = {ARTICLE},
 notes = {cited By 35},
 private_publication = {false},
 abstract = {GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Lowry, Jason A. and Mackay, Joel P.},
 doi = {10.1016/j.biocel.2005.06.017},
 journal = {The International Journal of Biochemistry & Cell Biology},
 number = {1}
}

@article{
 title = {The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7},
 type = {article},
 year = {2016},
 keywords = {MTA1,NuRD complex,RBBP4,chromatin,protein structure,transcription regulation},
 pages = {1472-1482},
 volume = {25},
 websites = {http://doi.wiley.com/10.1002/pro.2943},
 month = {8},
 id = {793f3c6b-8404-3b6e-9502-bf9b0357d1ca},
 created = {2020-12-17T05:29:59.209Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:59.209Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Schmidberger2016},
 source_type = {ARTICLE},
 notes = {cited By 4},
 private_publication = {false},
 abstract = {The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.},
 bibtype = {article},
 author = {Schmidberger, Jason W. and Sharifi Tabar, Mehdi and Torrado, Mario and Silva, Ana P. G. and Landsberg, Michael J. and Brillault, Lou and AlQarni, Saad and Zeng, Yi Cheng and Parker, Benjamin L. and Low, Jason K. K. and Mackay, Joel P.},
 doi = {10.1002/pro.2943},
 journal = {Protein Science},
 number = {8}
}

@article{
 title = {Two-state conformational equilibrium in the Par-4 leucine zipper domain},
 type = {article},
 year = {2010},
 keywords = {Circular dichroism,Leucine zipper,Prostate apoptosis response factor 4,Solution NMR spectroscopy},
 pages = {n/a-n/a},
 volume = {78},
 websites = {http://doi.wiley.com/10.1002/prot.22752},
 month = {4},
 day = {15},
 id = {90ab4173-c600-3c15-be8a-4dcbdfc4ed4d},
 created = {2020-12-17T05:29:59.280Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T05:29:59.280Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Schwalbe2010},
 source_type = {ARTICLE},
 notes = {cited By 5},
 private_publication = {false},
 abstract = {Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.},
 bibtype = {article},
 author = {Schwalbe, Martin and Dutta, Kaushik and Libich, David S. and Venugopal, Hariprasad and Claridge, Jolyon K. and Gell, David A. and Mackay, Joel P. and Edwards, Patrick J. B. and Pascal, Steven M.},
 doi = {10.1002/prot.22752},
 journal = {Proteins: Structure, Function, and Bioinformatics},
 number = {11}
}

@article{
 title = {Cyclic peptides can engage a single binding pocket through highly divergent modes},
 type = {article},
 year = {2020},
 keywords = {BET bromodomain inhibition,BRD3,BRD4,De novo cyclic peptides,Structural biology},
 volume = {117},
 id = {6636c6a0-e2f0-3aea-af8c-7865a5f17fcf},
 created = {2020-12-17T07:26:41.435Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2020-12-17T07:26:41.435Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {© 2020 National Academy of Sciences. All rights reserved. Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.},
 bibtype = {article},
 author = {Patel, K. and Walport, L.J. and Walshe, J.L. and Solomon, P.D. and Low, J.K.K. and Tran, D.H. and Mouradian, K.S. and Silva, A.P.G. and Wilkinson-White, L. and Norman, A. and Franck, C. and Matthews, J.M. and Mitchell Guss, J. and Payne, R.J. and Passioura, T. and Suga, H. and Mackay, J.P.},
 doi = {10.1073/pnas.2003086117},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {43}
}

@article{
 title = {Site-selective photocatalytic functionalization of peptides and proteins at selenocysteine},
 type = {article},
 year = {2022},
 volume = {13},
 id = {659f1a34-764d-3452-8843-3347bb2c9170},
 created = {2023-01-10T01:43:41.173Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:41.173Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The importance of modified peptides and proteins for applications in drug discovery, and for illuminating biological processes at the molecular level, is fueling a demand for efficient methods that facilitate the precise modification of these biomolecules. Herein, we describe the development of a photocatalytic method for the rapid and efficient dimerization and site-specific functionalization of peptide and protein diselenides. This methodology, dubbed the photocatalytic diselenide contraction, involves irradiation at 450 nm in the presence of an iridium photocatalyst and a phosphine and results in rapid and clean conversion of diselenides to reductively stable selenoethers. A mechanism for this photocatalytic transformation is proposed, which is supported by photoluminescence spectroscopy and density functional theory calculations. The utility of the photocatalytic diselenide contraction transformation is highlighted through the dimerization of selenopeptides, and by the generation of two families of protein conjugates via the site-selective modification of calmodulin containing the 21st amino acid selenocysteine, and the C-terminal modification of a ubiquitin diselenide.},
 bibtype = {article},
 author = {Dowman, L.J. and Kulkarni, S.S. and Alegre-Requena, J.V. and Giltrap, A.M. and Norman, A.R. and Sharma, A. and Gallegos, L.C. and Mackay, A.S. and Welegedara, A.P. and Watson, E.E. and Paton, R.S. and Payne, R.J.},
 doi = {10.1038/s41467-022-34530-z},
 journal = {Nature Communications},
 number = {1}
}

@article{
 title = {The role of auxiliary domains in modulating CHD4 activity suggests mechanistic commonality between enzyme families},
 type = {article},
 year = {2022},
 volume = {13},
 id = {76602a66-c88f-3ef1-8f5b-ae0a77f5e423},
 created = {2023-01-10T01:43:42.100Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:42.100Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {CHD4 is an essential, widely conserved ATP-dependent translocase that is also a broad tumour dependency. In common with other SF2-family chromatin remodelling enzymes, it alters chromatin accessibility by repositioning histone octamers. Besides the helicase and adjacent tandem chromodomains and PHD domains, CHD4 features 1000 residues of N- and C-terminal sequence with unknown structure and function. We demonstrate that these regions regulate CHD4 activity through different mechanisms. An N-terminal intrinsically disordered region (IDR) promotes remodelling integrity in a manner that depends on the composition but not sequence of the IDR. The C-terminal region harbours an auto-inhibitory region that contacts the helicase domain. Auto-inhibition is relieved by a previously unrecognized C-terminal SANT-SLIDE domain split by ~150 residues of disordered sequence, most likely by binding of this domain to substrate DNA. Our data shed light on CHD4 regulation and reveal strong mechanistic commonality between CHD family members, as well as with ISWI-family remodellers.},
 bibtype = {article},
 author = {Zhong, Y. and Moghaddas Sani, H. and Paudel, B.P. and Low, J.K.K. and Silva, A.P.G. and Mueller, S. and Deshpande, C. and Panjikar, S. and Reid, X.J. and Bedward, M.J. and van Oijen, A.M. and Mackay, J.P.},
 doi = {10.1038/s41467-022-35002-0},
 journal = {Nature Communications},
 number = {1}
}

@article{
 title = {Mucosal TLR2-activating protein-based vaccination induces potent pulmonary immunity and protection against SARS-CoV-2 in mice},
 type = {article},
 year = {2022},
 volume = {13},
 id = {817b23fa-ab01-305f-8861-84807b7e3327},
 created = {2023-01-10T01:43:43.018Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:43.018Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Current vaccines against SARS-CoV-2 substantially reduce mortality, but protection against infection is less effective. Enhancing immunity in the respiratory tract, via mucosal vaccination, may provide protection against infection and minimise viral spread. Here, we report testing of a subunit vaccine in mice, consisting of SARS-CoV-2 Spike protein with a TLR2-stimulating adjuvant (Pam2Cys), delivered to mice parenterally or mucosally. Both routes of vaccination induce substantial neutralising antibody (nAb) titres, however, mucosal vaccination uniquely generates anti-Spike IgA, increases nAb in the serum and airways, and increases lung CD4+ T-cell responses. TLR2 is expressed by respiratory epithelia and immune cells. Using TLR2 deficient chimeric mice, we determine that TLR2 expression in either compartment facilitates early innate responses to mucosal vaccination. By contrast, TLR2 on hematopoietic cells is essential for optimal lung-localised, antigen-specific responses. In K18-hACE2 mice, vaccination provides complete protection against disease and sterilising lung immunity against SARS-CoV-2, with a short-term non-specific protective effect from mucosal Pam2Cys alone. These data support mucosal vaccination as a strategy to improve protection in the respiratory tract against SARS-CoV-2 and other respiratory viruses.},
 bibtype = {article},
 author = {Ashhurst, A.S. and Johansen, M.D. and Maxwell, J.W.C. and Stockdale, S. and Ashley, C.L. and Aggarwal, A. and Siddiquee, R. and Miemczyk, S. and Nguyen, D.H. and Mackay, J.P. and Payne, R.J. and Britton, W.J.},
 doi = {10.1038/s41467-022-34297-3},
 journal = {Nature Communications},
 number = {1}
}

@article{
 title = {RNA inhibits dMi-2/CHD4 chromatin binding and nucleosome remodeling},
 type = {article},
 year = {2022},
 volume = {39},
 id = {d99d4ee9-901c-3e7f-9a93-d47a33ffacef},
 created = {2023-01-10T01:43:43.942Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:43.942Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The ATP-dependent nucleosome remodeler Mi-2/CHD4 broadly modulates chromatin landscapes to repress transcription and to maintain genome integrity. Here we use individual nucleotide resolution crosslinking and immunoprecipitation (iCLIP) to show that Drosophila Mi-2 associates with thousands of mRNA molecules in vivo. Biochemical data reveal that recombinant dMi-2 preferentially binds to G-rich RNA molecules using two intrinsically disordered regions of unclear function. Pharmacological inhibition of transcription and RNase digestion approaches establish that RNA inhibits the association of dMi-2 with chromatin. We also show that RNA inhibits dMi-2-mediated nucleosome mobilization by competing with the nucleosome substrate. Importantly, this activity is shared by CHD4, the human homolog of dMi-2, strongly suggesting that RNA-mediated regulation of remodeler activity is an evolutionary conserved mechanism. Our data support a model in which RNA serves to protect actively transcribed regions of the genome from dMi-2/CHD4-mediated establishment of repressive chromatin structures.},
 bibtype = {article},
 author = {Ullah, I. and Thölken, C. and Zhong, Y. and John, M. and Rossbach, O. and Lenz, J. and Gößringer, M. and Nist, A. and Albert, L. and Stiewe, T. and Mackay, J.P. and Brehm, A.},
 doi = {10.1016/j.celrep.2022.110895},
 journal = {Cell Reports},
 number = {9}
}

@article{
 title = {Mechanism of Bloom syndrome complex assembly required for double Holliday junction dissolution and genome stability},
 type = {article},
 year = {2022},
 volume = {119},
 id = {eea6affc-fd8d-3936-9453-941dacf51a3d},
 created = {2023-01-10T01:43:44.899Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:44.899Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The RecQ-like helicase BLM cooperates with topoisomerase IIIα, RMI1, and RMI2 in a heterotetrameric complex (the “Bloom syndrome complex”) for dissolution of double Holliday junctions, key intermediates in homologous recombination. Mutations in any component of the Bloom syndrome complex can cause genome instability and a highly cancer-prone disorder called Bloom syndrome. Some heterozygous carriers are also predisposed to breast cancer. To understand how the activities of BLM helicase and topoisomerase IIIα are coupled, we purified the active four-subunit complex. Chemical cross-linking and mass spectrometry revealed a unique architecture that links the helicase and topoisomerase domains. Using biochemical experiments, we demonstrated dimerization mediated by the N terminus of BLM with a 2:2:2:2 stoichiometry within the Bloom syndrome complex. We identified mutations that independently abrogate dimerization or association of BLM with RMI1, and we show that both are dysfunctional for dissolution using in vitro assays and cause genome instability and synthetic lethal interactions with GEN1/MUS81 in cells. Truncated BLM can also inhibit the activity of full-length BLM in mixed dimers, suggesting a putative mechanism of dominant-negative action in carriers of BLM truncation alleles. Our results identify critical molecular determinants of Bloom syndrome complex assembly required for double Holliday junction dissolution and maintenance of genome stability.},
 bibtype = {article},
 author = {Hodson, C. and Low, J.K.K. and van Twest, S. and Jones, S.E. and Swuec, P. and Murphy, V. and Tsukada, K. and Fawkes, M. and Bythell-Douglas, R. and Davies, A. and Costa, A. and Deans, A.J.},
 doi = {10.1073/pnas.2109093119},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {6}
}

@article{
 title = {Molecular architecture of nucleosome remodeling and deacetylase sub-complexes by integrative structure determination},
 type = {article},
 year = {2022},
 volume = {31},
 id = {eef4f63c-8daa-3623-b17b-a5cf28db9aee},
 created = {2023-01-10T01:43:45.810Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:45.810Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a chromatin-modifying assembly that regulates gene expression and DNA damage repair. Despite its importance, limited structural information describing the complete NuRD complex is available and a detailed understanding of its mechanism is therefore lacking. Drawing on information from SEC-MALLS, DIA-MS, XLMS, negative-stain EM, X-ray crystallography, NMR spectroscopy, secondary structure predictions, and homology models, we applied Bayesian integrative structure determination to investigate the molecular architecture of three NuRD sub-complexes: MTA1-HDAC1-RBBP4, MTA1N-HDAC1-MBD3GATAD2CC, and MTA1-HDAC1-RBBP4-MBD3-GATAD2A [nucleosome deacetylase (NuDe)]. The integrative structures were corroborated by examining independent crosslinks, cryo-EM maps, biochemical assays, known cancer-associated mutations, and structure predictions from AlphaFold. The robustness of the models was assessed by jack-knifing. Localization of the full-length MBD3, which connects the deacetylase and chromatin remodeling modules in NuRD, has not previously been possible; our models indicate two different locations for MBD3, suggesting a mechanism by which MBD3 in the presence of GATAD2A asymmetrically bridges the two modules in NuRD. Further, our models uncovered three previously unrecognized subunit interfaces in NuDe: HDAC1C-MTA1BAH, MTA1BAH-MBD3MBD, and HDAC160–100-MBD3MBD. Our approach also allowed us to localize regions of unknown structure, such as HDAC1C and MBD3IDR, thereby resulting in the most complete and robustly cross-validated structural characterization of these NuRD sub-complexes so far.},
 bibtype = {article},
 author = {Arvindekar, S. and Jackman, M.J. and Low, J.K.K. and Landsberg, M.J. and Mackay, J.P. and Viswanath, S.},
 doi = {10.1002/pro.4387},
 journal = {Protein Science},
 number = {9}
}

@article{
 title = {TNP Analogues Inhibit the Virulence Promoting IP<inf>3-4</inf> Kinase Arg1 in the Fungal Pathogen Cryptococcus neoformans},
 type = {article},
 year = {2022},
 volume = {12},
 id = {9cbc3fce-c9c1-35f8-ba6d-09bac2420d29},
 created = {2023-01-10T01:43:46.835Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:46.835Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {New antifungals with unique modes of action are urgently needed to treat the increasing global burden of invasive fungal infections. The fungal inositol polyphosphate kinase (IPK) pathway, comprised of IPKs that convert IP3 to IP8, provides a promising new target due to its impact on multiple, critical cellular functions and, unlike in mammalian cells, its lack of redundancy. Nearly all IPKs in the fungal pathway are essential for virulence, with IP3-4 kinase (IP3-4K) the most critical. The dibenzylaminopurine compound, N2-(m-trifluorobenzylamino)-N6-(p-nitrobenzylamino)purine (TNP), is a commercially available inhibitor of mammalian IPKs. The ability of TNP to be adapted as an inhibitor of fungal IP3-4K has not been investigated. We purified IP3-4K from the human pathogens, Cryptococcus neoformans and Candida albicans, and optimised enzyme and surface plasmon resonance (SPR) assays to determine the half inhibitory concentration (IC50) and binding affinity (KD), respectively, of TNP and 38 analogues. A novel chemical route was developed to efficiently prepare TNP analogues. TNP and its analogues demonstrated inhibition of recombinant IP3-4K from C. neoformans (CnArg1) at low µM IC50s, but not IP3-4K from C. albicans (CaIpk2) and many analogues exhibited selectivity for CnArg1 over the human equivalent, HsIPMK. Our results provide a foundation for improving potency and selectivity of the TNP series for fungal IP3-4K.},
 bibtype = {article},
 author = {Desmarini, D. and Truong, D. and Wilkinson-White, L. and Desphande, C. and Torrado, M. and Mackay, J.P. and Matthews, J.M. and Sorrell, T.C. and Lev, S. and Thompson, P.E. and Thompson, P.E. and Djordjevic, J.T.},
 doi = {10.3390/biom12101526},
 journal = {Biomolecules},
 number = {10}
}

@article{
 title = {Unique protein interaction networks define the chromatin remodelling module of the NuRD complex},
 type = {article},
 year = {2022},
 pages = {199-214},
 volume = {289},
 id = {a6268275-0bc8-3002-bce9-6d7a08deb3b1},
 created = {2023-01-10T01:43:47.765Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:47.765Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The combination of four proteins and their paralogues including MBD2/3, GATAD2A/B, CDK2AP1 and CHD3/4/5, which we refer to as the MGCC module, form the chromatin remodelling module of the nucleosome remodelling and deacetylase (NuRD) complex. To date, mechanisms by which the MGCC module acquires paralogue-specific function and specificity have not been addressed. Understanding the protein–protein interaction (PPI) network of the MGCC subunits is essential for defining underlying mechanisms of gene regulation. Therefore, using pulldown followed by mass spectrometry analysis (PD-MS), we report a proteome-wide interaction network of the MGCC module in a paralogue-specific manner. Our data also demonstrate that the disordered C-terminal region of CHD3/4/5 is a gateway to incorporate remodelling activity into both ChAHP (CHD4, ADNP, HP1γ) and NuRD complexes in a mutually exclusive manner. We define a short aggregation-prone region (APR) within the C-terminal segment of GATAD2B that is essential for the interaction of CHD4 and CDK2AP1 with the NuRD complex. Finally, we also report an association of CDK2AP1 with the nuclear receptor co-repressor (NCOR) complex. Overall, this study provides insight into the possible mechanisms through which the MGCC module can achieve specificity and diverse biological functions.},
 bibtype = {article},
 author = {Sharifi Tabar, M. and Giardina, C. and Feng, Y. and Francis, H. and Moghaddas Sani, H. and Low, J.K.K. and Mackay, J.P. and Bailey, C.G. and Rasko, J.E.J.},
 doi = {10.1111/febs.16112},
 journal = {FEBS Journal},
 number = {1}
}

@article{
 title = {BET-Family Bromodomains Can Recognize Diacetylated Sequences from Transcription Factors Using a Conserved Mechanism},
 type = {article},
 year = {2021},
 pages = {648-662},
 volume = {60},
 id = {50cd9f3e-3d6d-35ea-b47c-1090ed52dff5},
 created = {2023-01-10T01:43:48.713Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:48.713Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Almost all eukaryotic proteins receive diverse post-translational modifications (PTMs) that modulate protein activity. Many histone PTMs are well characterized, heavily influence gene regulation, and are often predictors of distinct transcriptional programs. Although our understanding of the histone PTM network has matured, much is yet to be understood about the roles of transcription factor (TF) PTMs, which might well represent a similarly complex and dynamic network of functional regulation. Members of the bromodomain and extra-terminal domain (BET) family of proteins recognize acetyllysine residues and relay the signals encoded by these modifications. Here, we have investigated the acetylation dependence of several functionally relevant BET-TF interactions in vitro using surface plasmon resonance, nuclear magnetic resonance, and X-ray crystallography. We show that motifs known to be acetylated in TFs E2F1 and MyoD1 can interact with all bromodomains of BRD2, BRD3, and BRD4. The interactions are dependent on diacetylation of the motifs and show a preference for the first BET bromodomain. Structural mapping of the interactions confirms a conserved mode of binding for the two TFs to the acetyllysine binding pocket of the BET bromodomains, mimicking that of other already established functionally important histone- and TF-BET interactions. We also examined a motif from the TF RelA that is known to be acetylated but were unable to observe any interaction, regardless of the acetylation state of the sequence. Our findings overall advance our understanding of BET-TF interactions and suggest a physical link between the important diacetylated motifs found in E2F1 and MyoD1 and the BET-family proteins.},
 bibtype = {article},
 author = {Patel, K. and Solomon, P.D. and Walshe, J.L. and Ford, D.J. and Wilkinson-White, L. and Payne, R.J. and Low, J.K.K. and Mackay, J.P.},
 doi = {10.1021/acs.biochem.0c00816},
 journal = {Biochemistry},
 number = {9}
}

@article{
 title = {The characterization of protein interactions-what, how and how much?},
 type = {article},
 year = {2021},
 pages = {12292-12307},
 volume = {50},
 id = {2d324f7d-025f-3a4a-af9f-b3ca4892be6b},
 created = {2023-01-10T01:43:49.626Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:49.626Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Protein interactions underlie most molecular events in biology. Many methods have been developed to identify protein partners, to measure the affinity with which these biomolecules interact and to characterise the structures of the complexes. Each approach has its own advantages and limitations, and it can be difficult for the newcomer to determine which methodology would best suit their system. This review provides an overview of many of the techniques most widely used to identify protein partners, assess stoichiometry and binding affinity, and determine low-resolution models for complexes. Key methods covered include: yeast two-hybrid analysis, affinity purification mass spectrometry and proximity labelling to identify partners; size-exclusion chromatography, scattering methods, native mass spectrometry and analytical ultracentrifugation to estimate stoichiometry; isothermal titration calorimetry, biosensors and fluorometric methods (including microscale thermophoresis, anisotropy/polarisation, resonance energy transfer, AlphaScreen, and differential scanning fluorimetry) to measure binding affinity; and crosslinking and hydrogen-deuterium exchange mass spectrometry to probe the structure of complexes. This journal is},
 bibtype = {article},
 author = {Walport, L.J. and Low, J.K.K. and Matthews, J.M. and MacKay, J.P.},
 doi = {10.1039/d1cs00548k},
 journal = {Chemical Society Reviews},
 number = {22}
}

@article{
 title = {The bromodomains of BET family proteins can recognize diacetylated histone H2A.Z},
 type = {article},
 year = {2021},
 pages = {464-476},
 volume = {30},
 id = {b502f967-797b-3fb0-8bb3-8923a5fa0bbd},
 created = {2023-01-10T01:43:50.588Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:50.588Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Chemical modifications of histone tails influence genome accessibility and the transcriptional state of eukaryotic cells. Lysine acetylation is one of the most common modifications and acetyllysine-binding bromodomains (BDs) provide a means for acetyllysine marks to be translated into meaningful cellular responses. Here, we have investigated the mechanism underlying the reported association between the Bromodomain and Extra Terminal (BET) family of BD proteins and the essential histone variant H2A.Z. We use NMR spectroscopy to demonstrate a physical interaction between the N-terminal tail of H2A.Z and the BDs of BRD2, BRD3, and BRD4, and show that the interaction is dependent on lysine acetylation in H2A.Z. The BDs preferentially engage a diacetylated H2A.Z-K4acK7ac motif that is reminiscent of sequences found in other biologically important BET BD target proteins, including histones and transcription factors. A H2A.Z-K7acK11ac motif can also bind BET BDs—with a preference for the second BD of each protein. Chemical shift perturbation mapping of the interactions, together with an X-ray crystal structure of BRD2-BD1 bound to H2A.Z-K4acK7ac, shows that H2A.Z binds the canonical AcK binding pocket of the BDs. This mechanism mirrors the conserved binding mode that is unique to the BET BDs, in which two acetylation marks are read simultaneously by a single BD. Our findings provide structural corroboration of biochemical and cell biological data that link H2A.Z and BET-family proteins, suggesting that the function of H2A.Z is enacted through interactions with these chromatin readers.},
 bibtype = {article},
 author = {Patel, K. and Solomon, P.D. and Walshe, J.L. and Low, J.K.K. and Mackay, J.P.},
 doi = {10.1002/pro.4006},
 journal = {Protein Science},
 number = {2}
}

@article{
 title = {Late-stage modification of peptides and proteins at cysteine with diaryliodonium salts},
 type = {article},
 year = {2021},
 pages = {14159-14166},
 volume = {12},
 id = {fe1847bf-a481-3fe2-9167-b70bf15b8ea0},
 created = {2023-01-10T01:43:51.522Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:51.522Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The modification of peptides and proteins has emerged as a powerful means to efficiently prepare high value bioconjugates for a range of applications in chemical biology and for the development of next-generation therapeutics. Herein, we report a novel method for the chemoselective late-stage modification of peptides and proteins at cysteine in aqueous buffer with suitably functionalised diaryliodonium salts, furnishing stable thioether-linked synthetic conjugates. The power of this new platform is showcased through the late-stage modification of the affibody zEGFR and the histone protein H2A.},
 bibtype = {article},
 author = {Byrne, S.A. and Bedding, M.J. and Corcilius, L. and Ford, D.J. and Zhong, Y. and Franck, C. and Larance, M. and Mackay, J.P. and Payne, R.J.},
 doi = {10.1039/d1sc03127a},
 journal = {Chemical Science},
 number = {42}
}

@article{
 title = {A single dose, BCG-adjuvanted COVID-19 vaccine provides sterilising immunity against SARS-CoV-2 infection},
 type = {article},
 year = {2021},
 volume = {6},
 id = {5f7017bd-f4d1-30e2-95a1-458fa2083a65},
 created = {2023-01-10T01:43:52.472Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:52.472Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Global control of COVID-19 requires broadly accessible vaccines that are effective against SARS-CoV-2 variants. In this report, we exploit the immunostimulatory properties of bacille Calmette-Guérin (BCG), the existing tuberculosis vaccine, to deliver a vaccination regimen with potent SARS-CoV-2-specific protective immunity. Combination of BCG with a stabilised, trimeric form of SARS-CoV-2 spike antigen promoted rapid development of virus-specific IgG antibodies in the blood of vaccinated mice, that was further augmented by the addition of alum. This vaccine formulation, BCG:CoVac, induced high-titre SARS-CoV-2 neutralising antibodies (NAbs) and Th1-biased cytokine release by vaccine-specific T cells, which correlated with the early emergence of T follicular helper cells in local lymph nodes and heightened levels of antigen-specific plasma B cells after vaccination. Vaccination of K18-hACE2 mice with a single dose of BCG:CoVac almost completely abrogated disease after SARS-CoV-2 challenge, with minimal inflammation and no detectable virus in the lungs of infected animals. Boosting BCG:CoVac-primed mice with a heterologous vaccine further increased SARS-CoV-2-specific antibody responses, which effectively neutralised B.1.1.7 and B.1.351 SARS-CoV-2 variants of concern. These findings demonstrate the potential for BCG-based vaccination to protect against major SARS-CoV-2 variants circulating globally.},
 bibtype = {article},
 author = {Counoupas, C. and Johansen, M.D. and Stella, A.O. and Nguyen, D.H. and Ferguson, A.L. and Aggarwal, A. and Bhattacharyya, N.D. and Grey, A. and Hutchings, O. and Patel, K. and Hansbro, P.M. and Triccas, J.A.},
 doi = {10.1038/s41541-021-00406-4},
 journal = {npj Vaccines},
 number = {1}
}

@article{
 title = {Purification of an insect juvenile hormone receptor complex enables insights into its post-translational phosphorylation},
 type = {article},
 year = {2021},
 volume = {297},
 id = {f039ef2e-e6b3-3564-a85c-0dc612c27e87},
 created = {2023-01-10T01:43:53.398Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:53.398Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Juvenile hormone (JH) plays vital roles in insect reproduction, development, and in many aspects of physiology. JH primarily acts at the gene-regulatory level through interaction with an intracellular receptor (JH receptor [JHR]), a ligand-activated complex of transcription factors consisting of the JH-binding protein methoprene-tolerant (MET) and its partner taiman (TAI). Initial studies indicated significance of post-transcriptional phosphorylation, subunit assembly, and nucleocytoplasmic transport of JHR in JH signaling. However, our knowledge of JHR regulation at the protein level remains rudimentary, partly because of the difficulty of obtaining purified and functional JHR proteins. Here, we present a method for high-yield expression and purification of JHR complexes from two insect species, the beetle T. castaneum and the mosquito Aedes aegypti. Recombinant JHR subunits from each species were coexpressed in an insect cell line using a baculovirus system. MET–TAI complexes were purified through affinity chromatography and anion exchange columns to yield proteins capable of binding both the hormonal ligand (JH III) and DNA bearing cognate JH-response elements. We further examined the beetle JHR complex in greater detail. Biochemical analyses and MS confirmed that T. castaneum JHR was a 1:1 heterodimer consisting of MET and Taiman proteins, stabilized by the JHR agonist ligand methoprene. Phosphoproteomics uncovered multiple phosphorylation sites in the MET protein, some of which were induced by methoprene treatment. Finally, we report a functional bipartite nuclear localization signal, straddled by phosphorylated residues, within the disordered C-terminal region of MET. Our present characterization of the recombinant JHR is an initial step toward understanding JHR structure and function.},
 bibtype = {article},
 author = {Jindra, M. and McKinstry, W.J. and Nebl, T. and Bittova, L. and Ren, B. and Shaw, J. and Phan, T. and Lu, L. and Low, J.K.K. and Mackay, J.P. and Lovrecz, G.O. and Hill, R.J.},
 doi = {10.1016/j.jbc.2021.101387},
 journal = {Journal of Biological Chemistry},
 number = {6}
}

@article{
 title = {Discovery of Cyclic Peptide Ligands to the SARS-CoV-2 Spike Protein Using mRNA Display},
 type = {article},
 year = {2021},
 pages = {1001-1008},
 volume = {7},
 id = {278be5c6-73f1-3344-9bd1-51ffab0169e6},
 created = {2023-01-10T01:43:54.314Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:54.314Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The COVID-19 pandemic, caused by SARS-CoV-2, has led to substantial morbidity, mortality, and disruption globally. Cellular entry of SARS-CoV-2 is mediated by the viral spike protein, and affinity ligands to this surface protein have the potential for applications as antivirals and diagnostic reagents. Here, we describe the affinity selection of cyclic peptide ligands to the SARS-CoV-2 spike protein receptor binding domain (RBD) from three distinct libraries (in excess of a trillion molecules each) by mRNA display. We identified six high affinity molecules with dissociation constants (KD) in the nanomolar range (15-550 nM) to the RBD. The highest affinity ligand could be used as an affinity reagent to detect the spike protein in solution by ELISA, and the cocrystal structure of this molecule bound to the RBD demonstrated that it binds to a cryptic binding site, displacing a β-strand near the C-terminus. Our findings provide key mechanistic insight into the binding of peptide ligands to the SARS-CoV-2 spike RBD, and the ligands discovered in this work may find future use as reagents for diagnostic applications.},
 bibtype = {article},
 author = {Norman, A. and Franck, C. and Christie, M. and Hawkins, P.M.E. and Patel, K. and Ashhurst, A.S. and Aggarwal, A. and Low, J.K.K. and Siddiquee, R. and Ashley, C.L. and Passioura, T. and Payne, R.J.},
 doi = {10.1021/acscentsci.0c01708},
 journal = {ACS Central Science},
 number = {6}
}

@article{
 title = {GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder},
 type = {article},
 year = {2020},
 pages = {878-888},
 volume = {22},
 id = {85308366-aa04-3a25-93ec-e3edc0ad78cc},
 created = {2023-01-10T01:43:55.280Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:55.280Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.},
 bibtype = {article},
 author = {Shieh, C. and Jones, N. and Vanle, B. and Au, M. and Huang, A.Y. and Silva, A.P.G. and Lee, H. and Douine, E.D. and Otero, M.G. and Choi, A. and Mackay, J.P. and Pierson, T.M.},
 doi = {10.1038/s41436-019-0747-z},
 journal = {Genetics in Medicine},
 number = {5}
}

@article{
 title = {Correction: Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system.(Applied Optics(2020)295(1898–1914)Doi: 10.1074/jbc.RA119.010679)},
 type = {article},
 year = {2020},
 pages = {6251},
 volume = {295},
 id = {9fc40b26-4df6-3126-8c8e-92fc8825b167},
 created = {2023-01-10T01:43:56.242Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:56.242Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Yichen Zhong's name was misspelled. The correct spelling is shown above.},
 bibtype = {article},
 author = {Werner, M.T. and Wang, H. and Hamagami, N. and Hsu, S.C. and Yano, J.A. and Stonestrom, A.J. and Behera, V. and Zhong, Y. and MacKay, J.P. and Blobel, G.A.},
 doi = {10.1074/jbc.AAC120.013771},
 journal = {Journal of Biological Chemistry},
 number = {18}
}

@article{
 title = {Cyclic peptides can engage a single binding pocket through highly divergent modes},
 type = {article},
 year = {2020},
 pages = {26728-26738},
 volume = {117},
 id = {9d68a322-e2a2-3b30-b21a-47d62f211339},
 created = {2023-01-10T01:43:57.164Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:57.164Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.},
 bibtype = {article},
 author = {Patel, K. and Walport, L.J. and Walshe, J.L. and Solomon, P.D. and Low, J.K.K. and Tran, D.H. and Mouradian, K.S. and Silva, A.P.G. and Wilkinson-White, L. and Norman, A. and Suga, H. and Mackay, J.P.},
 doi = {10.1073/pnas.2003086117},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {43}
}

@article{
 title = {Semisynthesis of an evasin from tick saliva reveals a critical role of tyrosine sulfation for chemokine binding and inhibition},
 type = {article},
 year = {2020},
 pages = {12657-12664},
 volume = {117},
 id = {b8c84666-fc07-3ad8-aaf9-d251ef3b0ee8},
 created = {2023-01-10T01:43:58.083Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:58.083Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Blood-feeding arthropods produce antiinflammatory salivary proteins called evasins that function through inhibition of chemokine-receptor signaling in the host. Herein, we show that the evasin ACA-01 from the Amblyomma cajennense tick can be posttranslationally sulfated at two tyrosine residues, albeit as a mixture of sulfated variants. Homogenously sulfated variants of the proteins were efficiently assembled via a semisynthetic native chemical ligation strategy. Sulfation significantly improved the binding affinity of ACA-01 for a range of proinflammatory chemokines and enhanced the ability of ACA-01 to inhibit chemokine signaling through cognate receptors. Comparisons of evasin sequences and structural data suggest that tyrosine sulfation serves as a receptor mimetic strategy for recognizing and suppressing the proinflammatory activity of a wide variety of mammalian chemokines. As such, the incorporation of this posttranslational modification (PTM) or mimics thereof into evasins may provide a strategy to optimize tick salivary proteins for antiinflammatory applications.},
 bibtype = {article},
 author = {Franck, C. and Foster, S.R. and Johansen-Leete, J. and Chowdhury, S. and Cielesh, M. and Bhusal, R.P. and Mackay, J.P. and Larance, M. and Stone, M.J. and Payne, R.J.},
 doi = {10.1073/pnas.2000605117},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {23}
}

@article{
 title = {A Novel Purification Procedure for Active Recombinant Human DPP4 and the Inability of DPP4 to Bind SARS-CoV-2},
 type = {article},
 year = {2020},
 volume = {25},
 id = {016baf4e-9b3e-39c6-87c2-be2e11c5cbaa},
 created = {2023-01-10T01:43:59.005Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:59.005Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Proteases catalyse irreversible posttranslational modifications that often alter a biological function of the substrate. The protease dipeptidyl peptidase 4 (DPP4) is a pharmacological target in type 2 diabetes therapy primarily because it inactivates glucagon-like protein-1. DPP4 also has roles in steatosis, insulin resistance, cancers and inflammatory and fibrotic diseases. In addition, DPP4 binds to the spike protein of the MERS virus, causing it to be the human cell surface receptor for that virus. DPP4 has been identified as a potential binding target of SARS-CoV-2 spike protein, so this question requires experimental investigation. Understanding protein structure and function requires reliable protocols for production and purification. We developed such strategies for baculovirus generated soluble recombinant human DPP4 (residues 29–766) produced in insect cells. Purification used differential ammonium sulphate precipitation, hydrophobic interaction chromatography, dye affinity chromatography in series with immobilised metal affinity chromatography, and ion-exchange chromatography. The binding affinities of DPP4 to the SARS-CoV-2 full-length spike protein and its receptor-binding domain (RBD) were measured using surface plasmon resonance and ELISA. This optimised DPP4 purification procedure yielded 1 to 1.8 mg of pure fully active soluble DPP4 protein per litre of insect cell culture with specific activity >30 U/mg, indicative of high purity. No specific binding between DPP4 and CoV-2 spike protein was detected by surface plasmon resonance or ELISA. In summary, a procedure for high purity high yield soluble human DPP4 was achieved and used to show that, unlike MERS, SARS-CoV-2 does not bind human DPP4.},
 bibtype = {article},
 author = {Xi, C.R. and Di Fazio, A. and Nadvi, N.A. and Patel, K. and Xiang, M.S.W. and Zhang, H.E. and Deshpande, C. and Low, J.K.K. and Wang, X.T. and Chen, Y. and Church, W.B. and Gorrell, M.D.},
 doi = {10.3390/MOLECULES25225392},
 journal = {Molecules},
 number = {22}
}

@article{
 title = {CHD4 slides nucleosomes by decoupling entry- and exit-side DNA translocation},
 type = {article},
 year = {2020},
 volume = {11},
 id = {5dc9783b-933a-3c63-9cd9-b4ae4e6fd62e},
 created = {2023-01-10T01:43:59.935Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:43:59.935Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Chromatin remodellers hydrolyse ATP to move nucleosomal DNA against histone octamers. The mechanism, however, is only partially resolved, and it is unclear if it is conserved among the four remodeller families. Here we use single-molecule assays to examine the mechanism of action of CHD4, which is part of the least well understood family. We demonstrate that the binding energy for CHD4-nucleosome complex formation—even in the absence of nucleotide—triggers significant conformational changes in DNA at the entry side, effectively priming the system for remodelling. During remodelling, flanking DNA enters the nucleosome in a continuous, gradual manner but exits in concerted 4–6 base-pair steps. This decoupling of entry- and exit-side translocation suggests that ATP-driven movement of entry-side DNA builds up strain inside the nucleosome that is subsequently released at the exit side by DNA expulsion. Based on our work and previous studies, we propose a mechanism for nucleosome sliding.},
 bibtype = {article},
 author = {Zhong, Y. and Paudel, B.P. and Ryan, D.P. and Low, J.K.K. and Franck, C. and Patel, K. and Bedward, M.J. and Torrado, M. and Payne, R.J. and van Oijen, A.M. and van Oijen, A.M. and Mackay, J.P.},
 doi = {10.1038/s41467-020-15183-2},
 journal = {Nature Communications},
 number = {1}
}

@article{
 title = {A heme-binding protein produced by Haemophilus haemolyticus inhibits non-typeable Haemophilus influenzae},
 type = {article},
 year = {2020},
 pages = {381-398},
 volume = {113},
 id = {9a9059a4-c040-3b6b-8ed2-e2bf25cedaa5},
 created = {2023-01-10T01:44:00.856Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:00.856Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Commensal bacteria serve as an important line of defense against colonisation by opportunisitic pathogens, but the underlying molecular mechanisms remain poorly explored. Here, we show that strains of a commensal bacterium, Haemophilus haemolyticus, make hemophilin, a heme-binding protein that inhibits growth of the opportunistic pathogen, non-typeable Haemophilus influenzae (NTHi) in culture. We purified the NTHi-inhibitory protein from H. haemolyticus and identified the hemophilin gene using proteomics and a gene knockout. An x-ray crystal structure of recombinant hemophilin shows that the protein does not belong to any of the known heme-binding protein folds, suggesting that it evolved independently. Biochemical characterisation shows that heme can be captured in the ferrous or ferric state, and with a variety of small heme-ligands bound, suggesting that hemophilin could function under a range of physiological conditions. Hemophilin knockout bacteria show a limited capacity to utilise free heme for growth. Our data suggest that hemophilin is a hemophore and that inhibition of NTHi occurs by heme starvation, raising the possibility that competition from hemophilin-producing H. haemolyticus could antagonise NTHi colonisation in the respiratory tract.},
 bibtype = {article},
 author = {Latham, R.D. and Torrado, M. and Atto, B. and Walshe, J.L. and Wilson, R. and Guss, J.M. and Mackay, J.P. and Tristram, S. and Gell, D.A.},
 doi = {10.1111/mmi.14426},
 journal = {Molecular Microbiology},
 number = {2}
}

@article{
 title = {Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system},
 type = {article},
 year = {2020},
 pages = {1898-1914},
 volume = {295},
 id = {53a92f38-e8f3-3c7c-9fcf-6b8b0bb9521a},
 created = {2023-01-10T01:44:01.787Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:01.787Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The widely expressed bromodomain and extraterminal motif (BET) proteins bromodomain-containing protein 2 (BRD2), BRD3, and BRD4 are multifunctional transcriptional regulators that bind acetylated chromatin via their conserved tandem bromodomains. Small molecules that target BET bromodomains are being tested for various diseases but typically do not discern between BET family members. Genomic distributions and protein partners of BET proteins have been described, but the basis for differences in BET protein function within a given lineage remains unclear. By establishing a gene knockout-rescue system in a Brd2-null erythroblast cell line, here we compared a series of mutant and chimeric BET proteins for their ability to modulate cell growth, differentiation, and gene expression. We found that the BET N-terminal halves bearing the bromodomains convey marked differences in protein stability but do not account for specificity in BET protein function. Instead, when BET proteins were expressed at comparable levels, their specificity was largely determined by the C-terminal half. Remarkably, a chimeric BET protein comprising the N-terminal half of the structurally similar short BRD4 isoform (BRD4S) and the C-terminal half of BRD2 functioned similarly to intact BRD2. We traced part of the BRD2-specific activity to a previously uncharacterized short segment predicted to harbor a coiled-coil (CC) domain. Deleting the CC segment impaired BRD2's ability to restore growth and differentiation, and the CC region functioned in conjunction with the adjacent ET domain to impart BRD2-like activity onto BRD4S. In summary, our results identify distinct BET protein domains that regulate protein turnover and biological activities.},
 bibtype = {article},
 author = {Werner, M.T. and Wang, H. and Hamagami, N. and Hsu, S.C. and Yano, J.A. and Stonestrom, A.J. and Behera, V. and Zong, Y. and Mackay, J.P. and Blobel, G.A.},
 doi = {10.1074/jbc.RA119.010679},
 journal = {Journal of Biological Chemistry},
 number = {7}
}

@article{
 title = {Thermostable small-molecule inhibitor of angiogenesis and vascular permeability that suppresses a pERK-FosB/ΔFosB-VCAM-1 axis},
 type = {article},
 year = {2020},
 volume = {6},
 id = {f819966b-5abf-3066-bb8c-9914a970f868},
 created = {2023-01-10T01:44:02.733Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:02.733Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Vascular permeability and angiogenesis underpin neovascular age-related macular degeneration and diabetic retinopathy. While anti-VEGF therapies are widely used clinically, many patients do not respond optimally, or at all, and small-molecule therapies are lacking. Here, we identified a dibenzoxazepinone BT2 that inhibits endothelial cell proliferation, migration, wound repair in vitro, network formation, and angiogenesis in mice bearing Matrigel plugs. BT2 interacts with MEK1 and inhibits ERK phosphorylation and the expression of FosB/ΔFosB, VCAM-1, and many genes involved in proliferation, migration, angiogenesis, and inflammation. BT2 reduced retinal vascular leakage following rat choroidal laser trauma and rabbit intravitreal VEGF-A165 administration. BT2 suppressed retinal CD31, pERK, VCAM-1, and VEGF-A165 expression. BT2 reduced retinal leakage in rats at least as effectively as aflibercept, a first-line therapy for nAMD/DR. BT2 withstands boiling or autoclaving and several months' storage at 22°C. BT2 is a new small-molecule inhibitor of vascular permeability and angiogenesis.},
 bibtype = {article},
 author = {Li, Y. and Alhendi, A.M.N. and Yeh, M.-C. and Elahy, M. and Santiago, F.S. and Deshpande, N.P. and Wu, B. and Chan, E. and Inam, S. and Prado-Lourenco, L. and Marcuccio, S.M. and Khachigian, L.M.},
 doi = {10.1126/sciadv.aaz7815},
 journal = {Science Advances},
 number = {31}
}

@article{
 title = {The Nucleosome Remodeling and Deacetylase Complex Has an Asymmetric, Dynamic, and Modular Architecture},
 type = {article},
 year = {2020},
 volume = {33},
 id = {a494d066-7bf4-3d91-b39e-2c81a3c17fb6},
 created = {2023-01-10T01:44:03.714Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:03.714Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Low et al. examine the architecture of the nucleosome remodeling and deacetylase complex. They define its stoichiometry, use cross-linking mass spectrometry to define subunit locations, and use electron microscopy to reveal large-scale dynamics. They also demonstrate that PWWP2A competes with MBD3 to sequester the HDAC-MTA-RBBP module from NuRD.SCOPUS_ABS_SEPARATORThe nucleosome remodeling and deacetylase (NuRD) complex is essential for metazoan development but has been refractory to biochemical analysis. We present an integrated analysis of the native mammalian NuRD complex, combining quantitative mass spectrometry, cross-linking, protein biochemistry, and electron microscopy to define the architecture of the complex. NuRD is built from a 2:2:4 (MTA, HDAC, and RBBP) deacetylase module and a 1:1:1 (MBD, GATAD2, and Chromodomain-Helicase-DNA-binding [CHD]) remodeling module, and the complex displays considerable structural dynamics. The enigmatic GATAD2 controls the asymmetry of the complex and directly recruits the CHD remodeler. The MTA-MBD interaction acts as a point of functional switching, with the transcriptional regulator PWWP2A competing with MBD for binding to the MTA-HDAC-RBBP subcomplex. Overall, our data address the long-running controversy over NuRD stoichiometry, provide imaging of the mammalian NuRD complex, and establish the biochemical mechanism by which PWWP2A can regulate NuRD composition.},
 bibtype = {article},
 author = {Low, J.K.K. and Silva, A.P.G. and Sharifi Tabar, M. and Torrado, M. and Webb, S.R. and Parker, B.L. and Sana, M. and Smits, C. and Schmidberger, J.W. and Brillault, L. and Landsberg, M.J. and Mackay, J.P.},
 doi = {10.1016/j.celrep.2020.108450},
 journal = {Cell Reports},
 number = {9}
}

@article{
 title = {Peppy: A virtual reality environment for exploring the principles of polypeptide structure},
 type = {article},
 year = {2020},
 pages = {157-168},
 volume = {29},
 id = {4e51d150-054e-3635-9211-b383f4822b0e},
 created = {2023-01-10T01:44:04.638Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:04.638Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.},
 bibtype = {article},
 author = {Doak, D.G. and Denyer, G.S. and Gerrard, J.A. and Mackay, J.P. and Allison, J.R.},
 doi = {10.1002/pro.3752},
 journal = {Protein Science},
 number = {1}
}

@article{
 title = {Correction: GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder (Genetics in Medicine, (2020), 10.1038/s41436-019-0747-z)},
 type = {article},
 year = {2020},
 pages = {822},
 volume = {22},
 id = {7ee81c43-2642-362b-af4d-d6af97c63161},
 created = {2023-01-10T01:44:05.579Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:05.579Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {An amendment to this paper has been published and can be accessed via a link at the top of the paper.},
 bibtype = {article},
 author = {Shieh, C. and Jones, N. and Vanle, B. and Au, M. and Huang, A.Y. and Silva, A.P.G. and Lee, H. and Douine, E.D. and Otero, M.G. and Choi, A. and Mackay, J.P. and Pierson, T.M.},
 doi = {10.1038/s41436-020-0760-2},
 journal = {Genetics in Medicine},
 number = {4}
}

@article{
 title = {The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G},
 type = {article},
 year = {2019},
 pages = {699-705},
 volume = {87},
 id = {eef77526-79b2-36df-bdf3-98887024df8f},
 created = {2023-01-10T01:44:06.506Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:06.506Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.},
 bibtype = {article},
 author = {Mohanty, B. and Hanson-Manful, P. and Finn, T.J. and Chambers, C.R. and McKellar, J.L.O. and Macindoe, I. and Helder, S. and Setiyaputra, S. and Zhong, Y. and Mackay, J.P. and Mackay, J.P. and Patrick, W.M.},
 doi = {10.1002/prot.25687},
 journal = {Proteins: Structure, Function and Bioinformatics},
 number = {8}
}

@article{
 title = {The stoichiometry and interactome of the Nucleosome Remodeling and Deacetylase (NuRD) complex are conserved across multiple cell lines},
 type = {article},
 year = {2019},
 pages = {2043-2061},
 volume = {286},
 id = {51b93072-bfe2-37ed-9321-35234e2dd662},
 created = {2023-01-10T01:44:07.415Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:07.415Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The nucleosome remodelling and deacetylase complex (NuRD) is a widely conserved regulator of gene expression. The determination of the subunit composition of the complex and identification of its binding partners are important steps towards understanding its architecture and function. The question of how these properties of the complex vary across different cell types has not been addressed in detail to date. Here, we set up a two-step purification protocol coupled to liquid chromatography-tandem mass spectrometry to assess NuRD composition and interaction partners in three different cancer cell lines, using label-free intensity-based absolute quantification (iBAQ). Our data indicate that the stoichiometry of the NuRD complex is preserved across our three different cancer cell lines. In addition, our interactome data suggest ZNF219 and SLC25A5 as possible interaction partners of the complex. To corroborate this latter finding, in vitro and cell-based pull-down experiments were carried out. These experiments indicated that ZNF219 can interact with RBBP4, GATAD2A/B and chromodomain helicase DNA binding 4, whereas SLC25A5 might interact with MTA2 and GATAD2A.},
 bibtype = {article},
 author = {Sharifi Tabar, M. and Mackay, J.P. and Low, J.K.K.},
 doi = {10.1111/febs.14800},
 journal = {FEBS Journal},
 number = {11}
}

@article{
 title = {Exploring the suitability of RanBP2-type Zinc Fingers for RNA-binding protein design},
 type = {article},
 year = {2019},
 volume = {9},
 id = {221c72ab-8395-34b9-89e2-2cc6f2d798f7},
 created = {2023-01-10T01:44:08.343Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:08.343Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Transcriptomes consist of several classes of RNA that have wide-ranging but often poorly described functions and the deregulation of which leads to numerous diseases. Engineering of functionalized RNA-binding proteins (RBPs) could therefore have many applications. Our previous studies suggested that the RanBP2-type Zinc Finger (ZF) domain is a suitable scaffold to investigate the design of single-stranded RBPs. In the present work, we have analyzed the natural sequence specificity of various members of the RanBP2-type ZF family and characterized the interaction with their target RNA. Surprisingly, our data showed that natural RanBP2-type ZFs with different RNA-binding residues exhibit a similar sequence specificity and therefore no simple recognition code can be established. Despite this finding, different discriminative abilities were observed within the family. In addition, in order to target a long RNA sequence and therefore gain in specificity, we generated a 6-ZF array by combining ZFs from the RanBP2-type family but also from different families, in an effort to achieve a wider target sequence repertoire. We showed that this chimeric protein recognizes its target sequence (20 nucleotides), both in vitro and in living cells. Altogether, our results indicate that the use of ZFs in RBP design remains attractive even though engineering of specificity changes is challenging.},
 bibtype = {article},
 author = {De Franco, S. and Vandenameele, J. and Brans, A. and Verlaine, O. and Bendak, K. and Damblon, C. and Matagne, A. and Segal, D.J. and Galleni, M. and Mackay, J.P. and Mackay, J.P. and Vandevenne, M.},
 doi = {10.1038/s41598-019-38655-y},
 journal = {Scientific Reports},
 number = {1}
}

@article{
 title = {The NuRD complex and macrocephaly associated neurodevelopmental disorders},
 type = {article},
 year = {2019},
 pages = {548-556},
 volume = {181},
 id = {03edfe18-463d-3f00-9f0b-586a1515ccc8},
 created = {2023-01-10T01:44:09.267Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:09.267Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.},
 bibtype = {article},
 author = {Pierson, T.M. and Otero, M.G. and Grand, K. and Choi, A. and Graham, J.M. and Young, J.I. and Mackay, J.P.},
 doi = {10.1002/ajmg.c.31752},
 journal = {American Journal of Medical Genetics, Part C: Seminars in Medical Genetics},
 number = {4}
}

@article{
 title = {Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding},
 type = {article},
 year = {2018},
 pages = {44-53},
 volume = {16},
 id = {086665b6-9995-388c-8525-8bf8e6dfb235},
 created = {2023-01-10T01:44:10.193Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:10.193Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.},
 bibtype = {article},
 author = {Mundell, S.J. and Rabbolini, D. and Gabrielli, S. and Chen, Q. and Aungraheeta, R. and Hutchinson, J.L. and Kilo, T. and Mackay, J. and Ward, C.M. and Stevenson, W. and Stevenson, W. and Morel-Kopp, M.-C.},
 doi = {10.1111/jth.13900},
 journal = {Journal of Thrombosis and Haemostasis},
 number = {1}
}

@article{
 title = {Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana},
 type = {article},
 year = {2018},
 pages = {167-176},
 volume = {8},
 id = {dd057f52-1171-342a-ba86-9f93ba6db989},
 created = {2023-01-10T01:44:11.110Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:11.110Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.},
 bibtype = {article},
 author = {Sani, H.M. and Hamzeh-Mivehroud, M. and Silva, A.P. and Walshe, J.L. and Abolghasem Mohammadi, S. and Rahbar-Shahrouziasl, M. and Abbasi, M. and Jamshidi, O. and Low, J.K.K. and Dastmalchi, S. and Dastmalchi, S. and Mackay, J.P.},
 doi = {10.15171/bi.2018.19},
 journal = {BioImpacts},
 number = {3}
}

@article{
 title = {PWWP2A binds distinct chromatin moieties and interacts with an MTA1-specific core NuRD complex},
 type = {article},
 year = {2018},
 volume = {9},
 id = {3ac8e87c-41a6-3ef1-859c-d226ac51ccaa},
 created = {2023-01-10T01:44:12.035Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:12.035Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Chromatin structure and function is regulated by reader proteins recognizing histone modifications and/or histone variants. We recently identified that PWWP2A tightly binds to H2A.Z-containing nucleosomes and is involved in mitotic progression and cranial–facial development. Here, using in vitro assays, we show that distinct domains of PWWP2A mediate binding to free linker DNA as well as H3K36me3 nucleosomes. In vivo, PWWP2A strongly recognizes H2A.Z-containing regulatory regions and weakly binds H3K36me3-containing gene bodies. Further, PWWP2A binds to an MTA1-specific subcomplex of the NuRD complex (M1HR), which consists solely of MTA1, HDAC1, and RBBP4/7, and excludes CHD, GATAD2 and MBD proteins. Depletion of PWWP2A leads to an increase of acetylation levels on H3K27 as well as H2A.Z, presumably by impaired chromatin recruitment of M1HR. Thus, this study identifies PWWP2A as a complex chromatin-binding protein that serves to direct the deacetylase complex M1HR to H2A.Z-containing chromatin, thereby promoting changes in histone acetylation levels.},
 bibtype = {article},
 author = {Link, S. and Spitzer, R.M.M. and Sana, M. and Torrado, M. and Völker-Albert, M.C. and Keilhauer, E.C. and Burgold, T. and Pünzeler, S. and Low, J.K.K. and Lindström, I. and Bartkuhn, M. and Hake, S.B.},
 doi = {10.1038/s41467-018-06665-5},
 journal = {Nature Communications},
 number = {1}
}

@book{
 title = {NMR spectroscopy in the analysis of protein-protein interactions},
 type = {book},
 year = {2018},
 source = {Modern Magnetic Resonance},
 pages = {2099-2132},
 id = {8844e241-63ec-3ea2-bdbe-1d62e7545c91},
 created = {2023-01-10T01:44:12.957Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:12.957Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.},
 bibtype = {book},
 author = {Gell, D.A. and Kwan, A.H. and Mackay, J.P.},
 doi = {10.1007/978-3-319-28388-3_121}
}

@article{
 title = {The BRD3 ET domain recognizes a short peptide motif through a mechanism that is conserved across chromatin remodelers and transcriptional regulators},
 type = {article},
 year = {2018},
 pages = {7160-7175},
 volume = {293},
 id = {312373ac-f639-35cd-8df1-cd3bdb1cbb6f},
 created = {2023-01-10T01:44:13.874Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:13.874Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Members of the bromodomain and extra-terminal domain (BET) family of proteins (bromodomain-containing (BRD) 2, 3, 4, and T) are widely expressed and highly conserved regulators of gene expression in eukaryotes. These proteins have been intimately linked to human disease, and more than a dozen clinical trials are currently underway to test BET-protein inhibitors as modulators of cancer. However, although it is clear that these proteins use their bromodomains to bind both histones and transcription factors bearing acetylated lysine residues, the molecular mechanisms by which BET family proteins regulate gene expression are not well defined. In particular, the functions of the other domains such as the ET domain have been less extensively studied. Here, we examine the properties of the ET domain of BRD3 as a protein/protein interaction module. Using a combination of pulldown and biophysical assays, we demonstrate that BRD3 binds to a range of chromatin-remodeling complexes, including the NuRD, BAF, and INO80 complexes, via a short linear “KIKL” motif in one of the complex subunits. NMR-based structural analysis revealed that, surprisingly, this mode of interaction is shared by the AF9 and ENL transcriptional coregulators that contain an acetyl-lysine-binding YEATS domain and regulate transcriptional elongation. This observation establishes a functional commonality between these two families of cancer-related transcriptional regulators. In summary, our data provide insight into the mechanisms by which BET family proteins might link chromatin acetylation to transcriptional outcomes and uncover an unexpected functional similarity between BET and YEATS family proteins.},
 bibtype = {article},
 author = {Wai, D.C.C. and Szyszka, T.N. and Campbell, A.E. and Kwong, C. and Lorna, E.W.-W. and Silva, A.P.G. and Low, J.K.K. and Kwan, A.H. and Gamsjaeger, R. and Chalmers, J.D. and Blobel, G.A. and Mackay, J.P.},
 doi = {10.1074/jbc.RA117.000678},
 journal = {Journal of Biological Chemistry},
 number = {19}
}

@article{
 title = {A Transcription Factor Addiction in Leukemia Imposed by the MLL Promoter Sequence},
 type = {article},
 year = {2018},
 pages = {970-981.e8},
 volume = {34},
 id = {cdabe452-f0bc-3b9a-a76b-3912d34763ca},
 created = {2023-01-10T01:44:14.809Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:14.809Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The Mixed Lineage Leukemia gene (MLL) is altered in leukemia by chromosomal translocations to produce oncoproteins composed of the MLL N-terminus fused to the C-terminus of a partner protein. Here, we used domain-focused CRISPR screening to identify ZFP64 as an essential transcription factor in MLL-rearranged leukemia. We show that the critical function of ZFP64 in leukemia is to maintain MLL expression via binding to the MLL promoter, which is the most enriched location of ZFP64 occupancy in the human genome. The specificity of ZFP64 for MLL is accounted for by an exceptional density of ZFP64 motifs embedded within the MLL promoter. These findings demonstrate how a sequence anomaly of an oncogene promoter can impose a transcriptional addiction in cancer.SCOPUS_ABS_SEPARATORLu et al. show that MLL-rearranged leukemia is addicted to the transcription factor ZFP64 due to direct regulation of MLL by ZFP64. The MLL promoter has an unusually high number of ZFP64 binding motifs and is the most enriched location of ZFP64 occupancy in the human genome.},
 bibtype = {article},
 author = {Lu, B. and Klingbeil, O. and Tarumoto, Y. and Somerville, T.D.D. and Huang, Y.-H. and Wei, Y. and Wai, D.C. and Low, J.K.K. and Milazzo, J.P. and Wu, X.S. and Shi, J. and Vakoc, C.R.},
 doi = {10.1016/j.ccell.2018.10.015},
 journal = {Cancer Cell},
 number = {6}
}

@article{
 title = {Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P},
 type = {article},
 year = {2018},
 pages = {1196-1209},
 volume = {19},
 id = {baa2b413-e43a-312a-9a1b-c88fce658272},
 created = {2023-01-10T01:44:15.759Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:15.759Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.},
 bibtype = {article},
 author = {Zhang, X. and Farah, N. and Rolston, L. and Ericsson, D.J. and Catanzariti, A.-M. and Bernoux, M. and Ve, T. and Bendak, K. and Chen, C. and Mackay, J.P. and Jones, D.A. and Kobe, B.},
 doi = {10.1111/mpp.12597},
 journal = {Molecular Plant Pathology},
 number = {5}
}

@article{
 title = {Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex},
 type = {article},
 year = {2017},
 pages = {4216-4232},
 volume = {284},
 id = {125673fd-8a9b-3e40-83fe-795ba612fa34},
 created = {2023-01-10T01:44:16.686Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:16.686Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.},
 bibtype = {article},
 author = {Torrado, M. and Low, J.K.K. and Silva, A.P.G. and Schmidberger, J.W. and Sana, M. and Sharifi Tabar, M. and Isilak, M.E. and Winning, C.S. and Kwong, C. and Bedward, M.J. and Shepherd, N.E. and Mackay, J.P.},
 doi = {10.1111/febs.14301},
 journal = {FEBS Journal},
 number = {24}
}

@article{
 title = {Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean},
 type = {article},
 year = {2017},
 pages = {1347-1355.e7},
 volume = {24},
 id = {f5f29eda-8c79-33fb-a24e-f144305413fb},
 created = {2023-01-10T01:44:17.671Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:17.671Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.},
 bibtype = {article},
 author = {Corcilius, L. and Hastwell, A.H. and Zhang, M. and Williams, J. and Mackay, J.P. and Gresshoff, P.M. and Ferguson, B.J. and Payne, R.J.},
 doi = {10.1016/j.chembiol.2017.08.014},
 journal = {Cell Chemical Biology},
 number = {11}
}

@article{
 title = {DCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression},
 type = {article},
 year = {2017},
 pages = {9901-9916},
 volume = {45},
 id = {5eedcada-99ed-3068-9c49-72ae47c1066a},
 created = {2023-01-10T01:44:18.596Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:18.596Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Distinct epigenomic profiles of histone marks have been associated with gene expression, but questions regarding the causal relationship remain. Here we investigated the activity of a broad collection of genomically targeted epigenetic regulators that could write epigenetic marks associated with a repressed chromatin state (G9A, SUV39H1, Krüppel-associated box (KRAB), DNMT3A as well as the first targetable versions of Ezh2 and Friend of GATA-1 (FOG1)). dCas9 fusions produced target gene repression over a range of 0- to 10-fold that varied by locus and cell type. dCpf1 fusions were unable to repress gene expression. The most persistent gene repression required the action of several effector domains; however, KRAB-dCas9 did not contribute to persistence in contrast to previous reports. A 'direct tethering' strategy attaching the Ezh2 methyltransferase enzyme to dCas9, as well as a 'recruitment' strategy attaching the N-terminal 45 residues of FOG1 to dCas9 to recruit the endogenous nucleosome remodeling and deacetylase complex, were both successful in targeted deposition of H3K27me3. Surprisingly, however, repression was not correlated with deposition of either H3K9me3 or H3K27me3. Our results suggest that so-called repressive histone modifications are not sufficient for gene repression. The easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation.},
 bibtype = {article},
 author = {O'Geen, H. and Ren, C. and Nicolet, C.M. and Perez, A.A. and Halmai, J. and Le, V.M. and MacKay, J.P. and Farnham, P.J. and Segal, D.J.},
 doi = {10.1093/nar/gkx578},
 journal = {Nucleic Acids Research},
 number = {17}
}

@article{
 title = {Promiscuous DNA-binding of a mutant zinc finger protein corrupts the transcriptome and diminishes cell viability},
 type = {article},
 year = {2017},
 pages = {1130-1143},
 volume = {45},
 id = {fc32d854-4e6d-3528-9963-88946b3e8087},
 created = {2023-01-10T01:44:19.547Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:19.547Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The rules of engagement between zinc finger transcription factors and DNA have been partly defined by in vitro DNA-binding and structural studies, but less is known about how these rules apply in vivo. Here, we demonstrate how a missense mutation in the second zinc finger of Krüppel-like factor-1 (KLF1) leads to degenerate DNA-binding specificity in vivo, resulting in ectopic transcription and anemia in the Nanmouse model. We employed ChIP-seq and 4sU-RNA-seq to identify aberrant DNA-binding events genome wide and ectopic transcriptional consequences of this binding. We confirmed novel sequence specificity of the mutant recombinant zinc finger domain by performing biophysical measurements ofinvitroDNA-binding affinity. Together, these results shed new light on the mechanisms by which missense mutations in DNA-binding domains of transcription factors can lead to autosomal dominant diseases.},
 bibtype = {article},
 author = {Gillinder, K.R. and Ilsley, M.D. and Nébor, D. and Sachidanandam, R. and Lajoie, M. and Magor, G.W. and Tallack, M.R. and Bailey, T. and Landsberg, M.J. and Mackay, J.P. and Bieker, J.J. and Perkins, A.C.},
 doi = {10.1093/nar/gkw1014},
 journal = {Nucleic Acids Research},
 number = {3}
}

@article{
 title = {IP<inf>3-4</inf> kinase Arg1 regulates cell wall homeostasis and surface architecture to promote clearance of Cryptococcus neoformans infection in a mouse model},
 type = {article},
 year = {2017},
 pages = {1833-1848},
 volume = {8},
 id = {2ce6221b-c29d-38b5-826d-bcfd3df72bba},
 created = {2023-01-10T01:44:20.463Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:20.463Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {We previously identified a series of inositol polyphosphate kinases (IPKs), Arg1, Ipk1, Kcs1 and Asp1, in the opportunistic fungal pathogen Cryptococcus neoformans. Using gene deletion analysis, we characterized Arg1, Ipk1 and Kcs1 and showed that they act sequentially to convert IP3 to PP-IP5 (IP7), a key metabolite promoting stress tolerance, metabolic adaptation and fungal dissemination to the brain. We have now directly characterized the enzymatic activity of Arg1, demonstrating that it is a dual specificity (IP3/IP4) kinase producing IP5. We showed previously that IP5 is further phosphorylated by Ipk1 to produce IP6, which is a substrate for the synthesis of PP-IP5 by Kcs1. Phenotypic comparison of the arg1Δ and kcs1Δ deletion mutants (both PP-IP5-deficient) reveals that arg1Δ has the most deleterious phenotype: while PP-IP5 is essential for metabolic and stress adaptation in both mutant strains, PP-IP5 is dispensable for virulence-associated functions such as capsule production, cell wall organization, and normal N-linked mannosylation of the virulence factor, phospholipase B1, as these phenotypes were defective only in arg1Δ. The more deleterious arg1Δ phenotype correlated with a higher rate of arg1Δ phagocytosis by human peripheral blood monocytes and rapid arg1Δ clearance from lung in a mouse model. This observation is in contrast to kcs1Δ, which we previously reported establishes a chronic, confined lung infection. In summary, we show that Arg1 is the most crucial IPK for cryptococcal virulence, conveying PP-IP5–dependent and novel PP-IP5–independent functions.},
 bibtype = {article},
 author = {Li, C. and Lev, S. and Desmarini, D. and Kaufman-Francis, K. and Saiardi, A. and Silva, A.P.G. and Mackay, J.P. and Thompson, P.E. and Sorrell, T.C. and Djordjevic, J.T.},
 doi = {10.1080/21505594.2017.1385692},
 journal = {Virulence},
 number = {8}
}

@article{
 title = {Whaddaya Know: A Guide to Uncertainty and Subjectivity in Structural Biology},
 type = {article},
 year = {2017},
 pages = {155-167},
 volume = {42},
 id = {cea96057-95a9-35d5-851d-39b4311c3896},
 created = {2023-01-10T01:44:21.385Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:21.385Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The methods of structural biology, while powerful, are technically complex. Although the Protein Data Bank (PDB) provides a repository that allows anyone to download any structure, many users would not appreciate the caveats that should be considered when examining a structure. Here, we describe several key uncertainties associated with the application of X-ray crystallography, NMR spectroscopy, single-particle electron microscopy (SPEM), and small-angle scattering (SAS) to biological macromolecules. The take-home message is that structures are not absolute truths – they are models that fit the experimental data and therefore have uncertainty and subjectivity associated with them. These uncertainties must be appreciated – careful reading of the associated paper, and any validation report provided by the structure database, is highly recommended.},
 bibtype = {article},
 author = {Mackay, J.P. and Landsberg, M.J. and Whitten, A.E. and Bond, C.S.},
 doi = {10.1016/j.tibs.2016.11.002},
 journal = {Trends in Biochemical Sciences},
 number = {2}
}

@article{
 title = {The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain},
 type = {article},
 year = {2016},
 pages = {4298-4314},
 volume = {428},
 id = {77987d3c-291d-39e9-97bc-2d692f6cd0c0},
 created = {2023-01-10T01:44:22.294Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:22.294Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.},
 bibtype = {article},
 author = {Mohanty, B. and Helder, S. and Silva, A.P.G. and Mackay, J.P. and Ryan, D.P.},
 doi = {10.1016/j.jmb.2016.08.028},
 journal = {Journal of Molecular Biology},
 number = {21}
}

@article{
 title = {Site-specific phosphorylation of tau inhibits amyloid-β toxicity in Alzheimer's mice},
 type = {article},
 year = {2016},
 pages = {904-908},
 volume = {354},
 id = {569c0c3a-417c-3254-a9b2-e49317df57d6},
 created = {2023-01-10T01:44:23.226Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:23.226Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Amyloid-β (Aβ) toxicity in Alzheimer's disease (AD) is considered to be mediated by phosphorylated tau protein. In contrast, we found that, at least in early disease, site-specific phosphorylation of tau inhibited Aβ toxicity. This specific tau phosphorylation was mediated by the neuronal p38 mitogen-activated protein kinase p38γ and interfered with postsynaptic excitotoxic signaling complexes engaged by Aβ. Accordingly, depletion of p38γ exacerbated neuronal circuit aberrations, cognitive deficits, and premature lethality in a mousemodel of AD, whereas increasing the activity of p38γ abolished these deficits. Furthermore, mimicking site-specific tau phosphorylation alleviated Aβ-induced neuronal death and offered protection from excitotoxicity. Our work provides insights into postsynaptic processes in AD pathogenesis and challenges a purely pathogenic role of tau phosphorylation in neuronal toxicity.},
 bibtype = {article},
 author = {Ittner, A. and Chua, S.W. and Bertz, J. and Volkerling, A. and Van Der Hoven, J. and Gladbach, A. and Przybyla, M. and Bi, M. and Van Hummel, A. and Stevens, C.H. and Ke, Y.D. and Ittner, L.M.},
 doi = {10.1126/science.aah6205},
 journal = {Science},
 number = {6314}
}

@article{
 title = {The zinc fingers of YY1 bind single-stranded RNA with low sequence specificity},
 type = {article},
 year = {2016},
 pages = {9153-9165},
 volume = {44},
 id = {448ec1cd-9b90-31ca-8df8-2d37eb36ba76},
 created = {2023-01-10T01:44:24.149Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:24.149Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Classical zinc fingers (ZFs) are traditionally considered to act as sequence-specific DNA-binding domains. More recently, classical ZFs have been recognised as potential RNA-binding modules, raising the intriguing possibility that classical-ZF transcription factors are involved in post-transcriptional gene regulation via direct RNA binding. To date, however, only one classical ZF-RNA complex, that involving TFIIIA, has been structurally characterised. Yin Yang-1 (YY1) is a multi-functional transcription factor involved in many regulatory processes, and binds DNA via four classical ZFs. Recent evidence suggests that YY1 also interacts with RNA, but the molecular nature of the interaction remains unknown. In the present work, we directly assess the ability of YY1 to bind RNA using in vitro assays. Systematic Evolution of Ligands by EXponential enrichment (SELEX) was used to identify preferred RNA sequences bound by the YY1 ZFs from a randomised library over multiple rounds of selection. However, a strong motif was not consistently recovered, suggesting that the RNA sequence selectivity of these domains is modest. YY1 ZF residues involved in binding to single-stranded RNA were identified by NMR spectroscopy and found to be largely distinct from the set of residues involved in DNA binding, suggesting that interactions between YY1 and ssRNA constitute a separate mode of nucleic acid binding. Our data are consistent with recent reports that YY1 can bind to RNA in a low-specificity, yet physiologically relevant manner.},
 bibtype = {article},
 author = {Wai, D.C.C. and Shihab, M. and Low, J.K.K. and Mackay, J.P.},
 doi = {10.1093/nar/gkw590},
 journal = {Nucleic Acids Research},
 number = {19}
}

@article{
 title = {Determinants of affinity and specificity in RNA-binding proteins},
 type = {article},
 year = {2016},
 pages = {83-91},
 volume = {38},
 id = {61fe4e83-b401-35f1-9433-78ccbe3d486b},
 created = {2023-01-10T01:44:25.084Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:25.084Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Emerging data suggest that the mechanisms by which RNA-binding proteins (RBPs) interact with RNA and the rules governing specificity might be substantially more complex than those underlying their DNA-binding counterparts. Even our knowledge of what constitutes the RNA-bound proteome is contentious; recent studies suggest that 10-30% of RBPs contain no known RNA-binding domain. Adding to this situation is a growing disconnect between the avalanche of identified interactions between proteins and long noncoding RNAs and the absence of biophysical data on these interactions. RNA-protein interactions are also at the centre of what might emerge as one of the biggest shifts in thinking about cell and molecular biology this century, following from recent reports of ribonucleoprotein complexes that drive reversible membrane-free phase separation events within the cell. Unexpectedly, low-complexity motifs are important in the formation of these structures. Here we briefly survey recent advances in our understanding of the specificity of RBPs.},
 bibtype = {article},
 author = {Helder, S. and Blythe, A.J. and Bond, C.S. and Mackay, J.P.},
 doi = {10.1016/j.sbi.2016.05.005},
 journal = {Current Opinion in Structural Biology}
}

@article{
 title = {The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7},
 type = {article},
 year = {2016},
 pages = {1472-1482},
 id = {a9a74730-2adb-3cee-87b3-9643769fab42},
 created = {2023-01-10T01:44:26.953Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:26.953Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.},
 bibtype = {article},
 author = {Schmidberger, J.W. and Sharifi Tabar, M. and Torrado, M. and Silva, A.P.G. and Landsberg, M.J. and Brillault, L. and AlQarni, S. and Zeng, Y.C. and Parker, B.L. and Low, J.K.K. and Low, J.K.K. and Mackay, J.P.},
 doi = {10.1002/pro.2943},
 journal = {Protein Science}
}

@article{
 title = {The N-terminal region of chromodomain helicase DNA-binding protein 4 (CHD4) is essential for activity and contains a high mobility group (HMG) box-like-domain that can bind poly(ADP-ribose)},
 type = {article},
 year = {2016},
 pages = {924-938},
 volume = {291},
 id = {ecf7907f-b86b-3542-ab23-e2a9a7ff5cc9},
 created = {2023-01-10T01:44:27.879Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:27.879Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Chromodomain Helicase DNA-binding protein 4 (CHD4) is a chromatin-remodeling enzyme that has been reported to regulate DNA-damage responses through its N-terminal region in a poly(ADP-ribose) polymerase-dependent manner. We have identified and determined the structure of a stable domain (CHD4-N) in this N-terminal region. The-fold consists of a four-α-helix bundle with structural similarity to the high mobility group box, a domain that is well known as a DNA binding module. We show that the CHD4-N domain binds with higher affinity to poly(ADP-ribose) than to DNA. We also show that the N-terminal region of CHD4, although not CHD4-N alone, is essential for full nucleosome remodeling activity and is important for localizing CHD4 to sites of DNA damage. Overall, these data build on our understanding of how CHD4-NuRD acts to regulate gene expression and participates in the DNA-damage response.},
 bibtype = {article},
 author = {Silva, A.P.G. and Ryan, D.P. and Galanty, Y. and Low, J.K.K. and Vandevenne, M. and Jackson, S.P. and Mackay, J.P.},
 doi = {10.1074/jbc.M115.683227},
 journal = {Journal of Biological Chemistry},
 number = {2}
}

@article{
 title = {CHD4 is a peripheral component of the nucleosome remodeling and deacetylase complex},
 type = {article},
 year = {2016},
 pages = {15853-15866},
 volume = {291},
 id = {7b57ef93-0acb-3843-a769-694abe573e25},
 created = {2023-01-10T01:44:28.938Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:28.938Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Chromatin remodeling enzymes act to dynamically regulate gene accessibility. In many cases, these enzymes function as large multicomponent complexes that in general comprise a central ATP-dependent Snf2 family helicase that is decorated with a variable number of regulatory subunits. The nucleosome remodeling and deacetylase (NuRD) complex, which is essential for normal development in higher organisms, is one such macromolecular machine. The NuRD complex comprises ∼10 subunits, including the histone deacetylases 1 and 2 (HDAC1 and HDAC2), and is defined by the presence of a CHD family remodeling enzyme, most commonly CHD4 (chromodomain helicase DNA-binding protein 4). The existing paradigm holds that CHD4 acts as the central hub upon which the complex is built. We show here that this paradigm does not, in fact, hold and that CHD4 is a peripheral component of the NuRD complex. A complex lacking CHD4 that has HDAC activity can exist as a stable species. The addition of recombinant CHD4 to this nucleosome deacetylase complex reconstitutes a NuRD complex with nucleosome remodeling activity. These data contribute to our understanding of the architecture of the NuRD complex.},
 bibtype = {article},
 author = {Low, J.K.K. and Webb, S.R. and Silva, A.P.G. and Saathoff, H. and Ryan, D.P. and Torrado, M. and Brofelth, M. and Parker, B.L. and Shepherd, N.E. and Mackay, J.P.},
 doi = {10.1074/jbc.M115.707018},
 journal = {Journal of Biological Chemistry},
 number = {30}
}

@article{
 title = {The binding of syndapin SH3 domain to Dynamin proline-rich domain involves short and long distance elements},
 type = {article},
 year = {2016},
 pages = {9411-9424},
 volume = {291},
 id = {34917dfc-3a3c-379e-a481-47c891a62429},
 created = {2023-01-10T01:44:29.922Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:29.922Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Dynamin is a GTPase that mediates vesicle fission during synaptic vesicle endocytosis. Its long C-terminal proline-rich domain contains 13 PXXP motifs, which orchestrate its interactions with multiple proteins. The SH3 domains of syndapin and endophilin bind the PXXP motifs called Site 2 and 3 (Pro-786-Pro-793) at the N-terminal end of the proline-rich domain, whereas the amphiphysin SH3 binds Site 9 (Pro-833-Pro-836) toward the C-terminal end. In some proteins, SH3/peptide interactions also involve short distance elements, which are 5-15 amino acid extensions flanking the central PXXP motif for high affinity binding. Here we found two previously unrecognized elements in the central and the C-terminal end of the dynamin proline-rich domain that account for a significant increase in syndapin binding affinity compared with a previously reported Site 2 and Site 3 PXXP peptide alone. The first new element (Gly-807-Gly-811) is short distance element on the C-terminal side of Site 2 PXXP, which might contact a groove identified under the RT loop of the SH3 domain. The second element (Arg-838-Pro-844) is located about 50 amino acids downstream of Site 2. These two elements provide additional specificity tothe syndapin SH3 domain outsideofthe well described polyproline-binding groove. Thus, the dynamin/syndapin interaction is mediated via a network of multiple contacts outside the core PXXP motif over a previously unrecognized extended region of the proline-rich domain. To our knowledge this is the first example among known SH3 interactions to involve spatially separated and extended long-range elements that combine to provide a higher affinity interaction. 7copy;2016 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Luo, L. and Xue, J. and Kwan, A. and Gamsjaeger, R. and Wielens, J. and Von Kleist, L. and Cubeddu, L. and Guo, Z. and Stow, J.L. and Parker, M.W. and Mackay, J.P. and Robinson, P.J.},
 doi = {10.1074/jbc.M115.703108},
 journal = {Journal of Biological Chemistry},
 number = {18}
}

@article{
 title = {<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of a C-terminal domain of human CHD1},
 type = {article},
 year = {2016},
 pages = {31-34},
 volume = {10},
 id = {b856848d-bf8e-37fe-813e-40d403bbd998},
 created = {2023-01-10T01:44:30.848Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:30.848Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Chromatin remodelling proteins are an essential family of eukaryotic proteins. They harness the energy from ATP hydrolysis and apply it to alter chromatin structure in order to regulate all aspects of genome biology. Chromodomain helicase DNA-binding protein 1 (CHD1) is one such remodelling protein that has specialised nucleosome organising abilities and is conserved across eukaryotes. CHD1 possesses a pair of tandem chromodomains that directly precede the core catalytic Snf2 helicase-like domain, and a C-terminal SANT-SLIDE DNA-binding domain. We have identified an additional conserved domain in the C-terminal region of CHD1. Here, we report the backbone and side chain resonance assignments for this domain from human CHD1 at pH 6.5 and 25 °C (BMRB No. 25638).},
 bibtype = {article},
 author = {Mohanty, B. and Silva, A.P.G. and Mackay, J.P. and Ryan, D.P.},
 doi = {10.1007/s12104-015-9631-1},
 journal = {Biomolecular NMR Assignments},
 number = {1}
}

@article{
 title = {The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein},
 type = {article},
 year = {2016},
 pages = {1710-1721},
 id = {7411c703-cf65-3456-a534-495980102c40},
 created = {2023-01-10T01:44:31.837Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:31.837Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.},
 bibtype = {article},
 author = {Blythe, A.J. and Yazar-Klosinski, B. and Webster, M.W. and Chen, E. and Vandevenne, M. and Bendak, K. and Mackay, J.P. and Hartzog, G.A. and Vrielink, A.},
 doi = {10.1002/pro.2976},
 journal = {Protein Science}
}

@article{
 title = {Paris-Trousseau thrombocytopenia is phenocopied by the autosomal recessive inheritance of a DNA-binding domain mutation in FLI1},
 type = {article},
 year = {2015},
 pages = {2027-2030},
 volume = {126},
 id = {9411fc17-2fe7-33f7-b1a3-3b0944dfec36},
 created = {2023-01-10T01:44:32.853Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:32.853Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Hemizygous deletion of a variable regionon chromosome 11q containing FLI1 causes an inherited platelet-related bleeding disorder in Paris-Trousseau thrombocytopenia and Jacobsen syndrome. These multisystem disorders are also characterized by heart anomalies, changes in facial structure, and intellectual disability. We have identified a consanguineous family with autosomal recessive inheritance of a bleeding disorder that mimics Paris-Trousseau thrombocytopenia but has no other features of the 11q23 deletion syndrome. Affected individuals inthis family have moderate thrombocytopenia; absent collagen-induced platelet aggregation; and large, fused α-granules in1%to 5% of circulating platelets. This phenotype was caused by a FLI1 homozygous c.970C>T-point mutation that predicts an arginine-to-tryptophan substitution in the conserved ETS DNA-binding domain of FLI1. This mutation caused a transcription defect at the promoter of known FLI1 target genes GP6, GP9, and ITGA2B, as measured by luciferase assay in HEK293 cells, and decreased the expression of these target proteins in affected members of the family as measured by Western blotting of platelet lysates. This kindred suggests abnormalitiesin FLI1 as causative of Paris-Trousseau thrombocytopenia and confirms the important role of FLI1 in normal platelet development.},
 bibtype = {article},
 author = {Stevenson, W.S. and Rabbolini, D.J. and Beutler, L. and Chen, Q. and Gabrielli, S. and Mackay, J.P. and Brighton, T.A. and Ward, C.M. and Morel-Kopp, M.-C.},
 doi = {10.1182/blood-2015-06-650887},
 journal = {Blood},
 number = {17}
}

@article{
 title = {Site directed nitroxide spin labeling of oligonucleotides for NMR and EPR studies},
 type = {article},
 year = {2015},
 pages = {813-819},
 volume = {71},
 id = {2ba001b8-f1dd-3136-90e2-56100f2c35ff},
 created = {2023-01-10T01:44:33.864Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:33.864Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Nitroxide labels are useful probes of biomolecule structure that can be detected by NMR and EPR spectroscopy. Although many methods exist for labeling oligonucleotides with nitroxides, most require reagents that are expensive or laborious to prepare. A simpler approach is described herein using commercially available phosphorothioate oligonucleotides and 2-(3-iodoacetamidomethyl)-PROXYL. We describe semi-optimized conditions for labeling DNA and RNA oligonucleotides and methodology for purifying and identifying these reagents by MALDI-MS. The nitroxide label showed some propensity to hydrolyze at high temperature and over prolonged periods at room temperature. Nitroxide-labeled DNA oligonucleotides gave characteristic EPR spectra and caused the disappearance of bound protein signals in 15N HSQC spectra consistent with the paramagnetic relaxation enhancement (PRE) effect.},
 bibtype = {article},
 author = {Shepherd, N.E. and Gamsjaeger, R. and Vandevenne, M. and Cubeddu, L. and Mackay, J.P.},
 doi = {10.1016/j.tet.2014.12.056},
 journal = {Tetrahedron},
 number = {5}
}

@article{
 title = {A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells},
 type = {article},
 year = {2015},
 pages = {960-965},
 volume = {23},
 id = {4e5561f0-05d0-3b6a-8a77-9e05d0b36591},
 created = {2023-01-10T01:44:34.787Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:34.787Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).},
 bibtype = {article},
 author = {Saathoff, H. and Brofelth, M. and Trinh, A. and Parker, B.L. and Ryan, D.P. and Low, J.K.K. and Webb, S.R. and Silva, A.P.G. and Mackay, J.P. and Shepherd, N.E.},
 doi = {10.1016/j.bmc.2015.01.023},
 journal = {Bioorganic and Medicinal Chemistry},
 number = {5}
}

@article{
 title = {The identification and structure of an N-terminal PR domain show that FOG1 is a member of the PRDM family of proteins},
 type = {article},
 year = {2014},
 volume = {9},
 id = {d4bd4375-43f0-3b26-8cf7-d8fffdd6c864},
 created = {2023-01-10T01:44:35.718Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:35.718Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {FOG1 is a transcriptional regulator that acts in concert with the hematopoietic master regulator GATA1 to coordinate the differentiation of platelets and erythrocytes. Despite considerable effort, however, the mechanisms through which FOG1 regulates gene expression are only partially understood. Here we report the discovery of a previously unrecognized domain in FOG1: a PR (PRD-BF1 and RIZ) domain that is distantly related in sequence to the SET domains that are found in many histone methyltransferases. We have used NMR spectroscopy to determine the solution structure of this domain, revealing that the domain shares close structural similarity with SET domains. Titration with S-adenosyl-L-homocysteine, the cofactor product synonymous with SET domain methyltransferase activity, indicated that the FOG PR domain is not, however, likely to function as a methyltransferase in the same fashion. We also sought to define the function of this domain using both pulldown experiments and gel shift assays. However, neither pulldowns from mammalian nuclear extracts nor yeast two-hybrid assays reproducibly revealed binding partners, and we were unable to detect nucleic-acid-binding activity in this domain using our high-diversity Pentaprobe oligonucleotides. Overall, our data demonstrate that FOG1 is a member of the PRDM (PR domain containing proteins, with zinc fingers) family of transcriptional regulators. The function of many PR domains, however, remains somewhat enigmatic for the time being. © 2014 Clifton et al.},
 bibtype = {article},
 author = {Clifton, M.K. and Westman, B.J. and Thong, S.Y. and O'Connell, M.R. and Webster, M.W. and Shepherd, N.E. and Quinlan, K.G. and Crossley, M. and Blobel, G.A. and Mackay, J.P.},
 doi = {10.1371/journal.pone.0106011},
 journal = {PLoS ONE},
 number = {8}
}

@article{
 title = {Ca2+-induced PRE-NMR changes in the troponin complex reveal the possessive nature of the cardiac isoform for its regulatory switch},
 type = {article},
 year = {2014},
 volume = {9},
 id = {c3b7b413-fdec-325f-9bcd-b242670e45a7},
 created = {2023-01-10T01:44:36.628Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:36.628Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The interaction between myosin and actin in cardiac muscle, modulated by the calcium (Ca2+) sensor Troponin complex (Tn), is a complex process which is yet to be fully resolved at the molecular level. Our understanding of how the binding of Ca2+ triggers conformational changes within Tn that are subsequently propagated through the contractile apparatus to initiate muscle activation is hampered by a lack of an atomic structure for the Ca2+-free state of the cardiac isoform. We have used paramagnetic relaxation enhancement (PRE)-NMR to obtain a description of the Ca2+-free state of cardiac Tn by describing the movement of key regions of the troponin I (cTnI) subunit upon the release of Ca2+ from Troponin C (cTnC). Site-directed spin-labeling was used to position paramagnetic spin labels in cTnI and the changes in the interaction between cTnI and cTnC subunits were then mapped by PRE-NMR. The functionally important regions of cTnI targeted in this study included the cTnC-binding N-region (cTnI57), the inhibitory region (cTnI143), and two sites on the regulatory switch region (cTnI151 and cTnI159). Comparison of 1H-15N-TROSY spectra of Ca2+-bound and free states for the spin labeled cTnCcTnI binary constructs demonstrated the release and modest movement of the cTnI switch region (,10 A ) away from the hydrophobic N-lobe of troponin C (cTnC) upon the removal of Ca2+. Our data supports a model where the non-bound regulatory switch region of cTnI is highly flexible in the absence of Ca2+ but remains in close vicinity to cTnC. We speculate that the close proximity of TnI to TnC in the cardiac complex is favourable for increasing the frequency of collisions between the N-lobe of cTnC and the regulatory switch region, counterbalancing the reduction in collision probability that results from the incomplete opening of the N-lobe of TnC that is unique to the cardiac isoform.},
 bibtype = {article},
 author = {Cordina, N.M. and Liew, C.K. and Potluri, P.R. and Curmi, P.M. and Fajer, P.G. and Logan, T.M. and Mackay, J.P. and Brown, L.J.},
 doi = {10.1371/journal.pone.0112976},
 journal = {PLoS ONE},
 number = {11}
}

@article{
 title = {Engineering specificity changes on a RanBP2 zinc finger that binds single-stranded RNA},
 type = {article},
 year = {2014},
 pages = {7848-7852},
 volume = {53},
 id = {efff0e09-c7d0-32a8-9607-835d587fd71a},
 created = {2023-01-10T01:44:37.599Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:37.599Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.},
 bibtype = {article},
 author = {Vandevenne, M. and O'Connell, M.R. and Helder, S. and Shepherd, N.E. and Matthews, J.M. and Kwan, A.H. and Segal, D.J. and Mackay, J.P.},
 doi = {10.1002/anie.201402980},
 journal = {Angewandte Chemie - International Edition},
 number = {30}
}

@article{
 title = {The structure of an LIM-Only protein 4 (LMO4) and Deformed Epidermal Autoregulatory Factor-1 (DEAF1) complex reveals a common mode of binding to LMO4},
 type = {article},
 year = {2014},
 volume = {9},
 id = {5973cfa4-e9ae-3e89-b66f-cf09132c9ab9},
 created = {2023-01-10T01:44:38.507Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:38.507Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {LIM-domain only protein 4 (LMO4) is a widely expressed protein with important roles in embryonic development and breast cancer. It has been reported to bind many partners, including the transcription factor Deformed epidermal autoregulatory factor-1 (DEAF1), with which LMO4 shares many biological parallels. We used yeast two-hybrid assays to show that DEAF1 binds both LIM domains of LMO4 and that DEAF1 binds the same face on LMO4 as two other LMO4-binding partners, namely LIM domain binding protein 1 (LDB1) and C-terminal binding protein interacting protein (CtIP/RBBP8). Mutagenic screening analysed by the same method, indicates that the key residues in the interaction lie in LMO4LIM2and the N-terminal half of the LMO4-binding domain in DEAF1. We generated a stable LMO4LIM2-DEAF1 complex and determined the solution structure of that complex. Although the LMO4-binding domain from DEAF1 is intrinsically disordered, it becomes structured on binding. The structure confirms that LDB1, CtIP and DEAF1 all bind to the same face on LMO4. LMO4 appears to form a hub in protein-protein interaction networks, linking numerous pathways within cells. Competitive binding for LMO4 therefore most likely provides a level of regulation between those different pathways.},
 bibtype = {article},
 author = {Joseph, S. and Kwan, A.H. and Stokes, P.H. and Mackay, J.P. and Cubeddu, L. and Matthews, J.M.},
 doi = {10.1371/journal.pone.0109108},
 journal = {PLoS ONE},
 number = {10}
}

@article{
 title = {Insight into the architecture of the NuRD complex: Structure of the RbAp48-MTA1 subcomplex},
 type = {article},
 year = {2014},
 pages = {21844-21855},
 volume = {289},
 id = {a0afaf4d-99c7-3fea-9a0a-a55fcb967c4a},
 created = {2023-01-10T01:44:39.545Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:39.545Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background: The NuRD complex controls gene expression through altering chromatin structure. Results: The MTA1-RbAp48 structure shows how the RbAp46/p48 histone chaperones are recruited to NuRD. Conclusion: The MTA subunits act as scaffolds for NuRD complex assembly. Significance: The MTA/RbAp48 interaction prevents binding of histone H4, which is crucial for understanding the role of the RbAp46/p48 chaperones in the complex. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Alqarni, S.S.M. and Murthy, A. and Zhang, W. and Przewloka, M.R. and Silva, A.P.G. and Watson, A.A. and Lejon, S. and Pei, X.Y. and Smits, A.H. and Kloet, S.L. and Mackay, J.P. and Laue, E.D.},
 doi = {10.1074/jbc.M114.558940},
 journal = {Journal of Biological Chemistry},
 number = {32}
}

@article{
 title = {Structure of the hemoglobin-isdh complex reveals the molecular basis of iron capture by staphylococcus aureus},
 type = {article},
 year = {2014},
 pages = {6728-6738},
 volume = {289},
 id = {8082b276-9693-3b97-b289-93ddb85c7c38},
 created = {2023-01-10T01:44:40.488Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:40.488Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background: IsdB and IsdH proteins from Staphylococcus aureus strip heme iron from human hemoglobin. Results: The IsdH·hemoglobin complex shows how globin-binding and heme-binding NEAT domains of IsdH cooperate to remove heme from both chains of hemoglobin. Conclusion: The supradomain architecture of IsdH confers activity by precisely positioning the heme acceptor domain. Significance: Multiple IsdH·hemoglobin interfaces may be targets for new antibiotics. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Dickson, C.F. and Kumar, K.K. and Jacques, D.A. and Malmirchegini, G.R. and Spirig, T. and Mackay, J.P. and Clubb, R.T. and Guss, J.M. and Gell, D.A.},
 doi = {10.1074/jbc.M113.545566},
 journal = {Journal of Biological Chemistry},
 number = {10}
}

@article{
 title = {Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5},
 type = {article},
 year = {2014},
 pages = {54-60},
 volume = {100},
 id = {79bfac53-430c-3f33-98e9-37db63509339},
 created = {2023-01-10T01:44:41.424Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:41.424Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.},
 bibtype = {article},
 author = {Blythe, A. and Gunasekara, S. and Walshe, J. and Mackay, J.P. and Hartzog, G.A. and Vrielink, A.},
 doi = {10.1016/j.pep.2014.05.005},
 journal = {Protein Expression and Purification}
}

@article{
 title = {Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1},
 type = {article},
 year = {2014},
 pages = {141-144},
 volume = {8},
 id = {a64344fa-3b32-37f0-b1e1-f7b7ad8cbfd8},
 created = {2023-01-10T01:44:42.330Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:42.330Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.},
 bibtype = {article},
 author = {Joseph, S. and Kwan, A.H.Y. and Mackay, J.P. and Cubeddu, L. and Matthews, J.M.},
 doi = {10.1007/s12104-013-9470-x},
 journal = {Biomolecular NMR Assignments},
 number = {1}
}

@article{
 title = {Trim58 Degrades Dynein and Regulates Terminal Erythropoiesis},
 type = {article},
 year = {2014},
 pages = {688-700},
 volume = {30},
 id = {c1ccb790-7e19-39e4-aaad-2c8dd2dc11ef},
 created = {2023-01-10T01:44:43.258Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:43.258Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {TRIM58 is an E3 ubiquitin ligase superfamily member implicated by genome-wide association studies to regulate human erythrocyte traits. Here, we show that Trim58 expression is induced during late erythropoiesis and that its depletion by small hairpin RNAs (shRNAs) inhibits the maturation of late-stage nucleated erythroblasts to anucleate reticulocytes. Imaging flow cytometry studies demonstrate that Trim58 regulates polarization and/or extrusion of erythroblast nuclei. Invitro, Trim58 directly binds and ubiquitinates the intermediate chain of the microtubule motor dynein. In cells, Trim58 stimulates proteasome-dependent degradation of the dynein holoprotein complex. During erythropoiesis, Trim58 expression, dynein loss, and enucleation occur concomitantly, and all are inhibited by Trim58 shRNAs. Dynein regulates nuclear positioning and microtubule organization, both of which undergo dramatic changes during erythroblast enucleation. Thus, we propose that Trim58 promotes this processby eliminating dynein. Our findings identify an erythroid-specific regulator of enucleation and elucidate a previously unrecognized mechanism for controlling dynein activity.},
 bibtype = {article},
 author = {Thom, C.S. and Traxler, E.A. and Khandros, E. and Nickas, J.M. and Zhou, O.Y. and Lazarus, J.E. and Silva, A.P.G. and Prabhu, D. and Yao, Y. and Aribeana, C. and Holzbaur, E.L.F. and Weiss, M.J.},
 doi = {10.1016/j.devcel.2014.07.021},
 journal = {Developmental Cell},
 number = {6}
}

@article{
 title = {Transcription factor seeks DNA - Cognate site preferred},
 type = {article},
 year = {2014},
 pages = {1370-1372},
 volume = {426},
 id = {1e075f30-6417-39c7-8b24-b13d5c28f504},
 created = {2023-01-10T01:44:44.178Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:44.178Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Mackay, J.},
 doi = {10.1016/j.jmb.2013.12.010},
 journal = {Journal of Molecular Biology},
 number = {7}
}

@article{
 title = {<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of an N-terminal domain of CHD4},
 type = {article},
 year = {2014},
 pages = {137-139},
 volume = {8},
 id = {cd0244ce-1944-3196-a041-f999ed20f6c3},
 created = {2023-01-10T01:44:45.086Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:45.086Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.},
 bibtype = {article},
 author = {Silva, A.P.G. and Kwan, A.H. and Mackay, J.P.},
 doi = {10.1007/s12104-013-9469-3},
 journal = {Biomolecular NMR Assignments},
 number = {1}
}

@article{
 title = {α-Hemoglobin-stabilizing Protein (AHSP) perturbs the proximal heme pocket of oxy-α-hemoglobin and weakens the iron-oxygen bond},
 type = {article},
 year = {2013},
 pages = {19986-20001},
 volume = {288},
 id = {d5cb8d14-4ed7-37da-a53b-1987811758ed},
 created = {2023-01-10T01:44:46.024Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:46.024Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {α-Hemoglobin (αHb)-stabilizing protein (AHSP) is a molecular chaperone that assists hemoglobin assembly. AHSP induces changes in αHb heme coordination, but how these changes are facilitated by interactions at the αHb·AHSP interface is not well understood. To address this question we have used NMR, x-ray absorption spectroscopy, and ligand binding measurements to probe αHb conformational changes induced by AHSP binding.NMRchemical shift analyses of free CO-αHb and CO- αHb·AHSP indicated that the seven helical elements of the native αHb structure are retained and that the heme Fe(II) remains coordinated to the proximal His-87 side chain. However, chemical shift differences revealed alterations of the F, G, and H helices and the heme pocket of CO-αHb bound to AHSP. Comparisons of iron-ligand geometry using extended x-ray absorption fine structure spectroscopy showed that AHSP binding induces a small 0.03 Å lengthening of the Fe-O2 bond, explaining previous reports that AHSP decreases αHb O2 affinity roughly 4-fold and promotes autooxidation due primarily to a 3-4-fold increase in the rate ofO2 dissociation. Pro-30 mutations diminished NMR chemical shift changes in the proximal heme pocket, restored normal O2 dissociation rate and equilibrium constants, and reduced O2-αHb autooxidation rates. Thus, the contacts mediated by Pro-30 in wild-type AHSP promote αHb autooxidation by introducing strain into the proximal heme pocket. As a chaperone, AHSP facilitates rapid assembly of αHb into Hb when βHb is abundant but diverts αHb to a redox resistant holding state when βHb is limiting. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Dickson, C.F. and Rich, A.M. and D'Avigdor, W.M.H. and Collins, D.A.T. and Lowry, J.A. and Mollan, T.L. and Khandros, E. and Olson, J.S. and Weiss, M.J. and MacKay, J.P. and Lay, P.A. and Gell, D.A.},
 doi = {10.1074/jbc.M112.437509},
 journal = {Journal of Biological Chemistry},
 number = {27}
}

@article{
 title = {Is there a telltale RH fingerprint in zinc fingers that recognizes methylated CpG dinucleotides?},
 type = {article},
 year = {2013},
 pages = {421-422},
 volume = {38},
 id = {4f05c721-c533-3f78-93e4-e5649cccf00b},
 created = {2023-01-10T01:44:46.983Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:46.983Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Mackay, J.P. and Segal, D.J. and Crossley, M.},
 doi = {10.1016/j.tibs.2013.06.005},
 journal = {Trends in Biochemical Sciences},
 number = {9}
}

@article{
 title = {A structural analysis of DNA binding by myelin transcription factor 1 double zinc fingers},
 type = {article},
 year = {2013},
 pages = {35180-35191},
 volume = {288},
 id = {87086ec5-3591-3e63-826a-b7a3fde32906},
 created = {2023-01-10T01:44:47.925Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:47.925Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background: Myelin transcription factor 1 (MyT1) contains seven similar zinc finger domains that bind DNA specifically. Results: A three-dimensional structural model explains how a double zinc finger unit is able to recognize DNA. Conclusion: DNA-binding residues are conserved among all MyT1 zinc fingers, suggesting an identical DNA binding mode. Significance: Determination of the molecular details of DNA interaction will be crucial in understanding MyT1 function. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Gamsjaeger, R. and O'Connell, M.R. and Cubeddu, L. and Shepherd, N.E. and Lowry, J.A. and Kwan, A.H. and Vandevenne, M. and Swanton, M.K. and Matthews, J.M. and Mackay, J.P.},
 doi = {10.1074/jbc.M113.482075},
 journal = {Journal of Biological Chemistry},
 number = {49}
}

@article{
 title = {New insights into DNA recognition by zinc fingers revealed by structural analysis of the oncoprotein ZNF217},
 type = {article},
 year = {2013},
 pages = {10616-10627},
 volume = {288},
 id = {406b142f-54cb-305c-8eaa-3b8866da36f3},
 created = {2023-01-10T01:44:48.847Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:48.847Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background: Classical zinc finger proteins are extremely abundant and interact with DNA using a well defined recognition code. Results: We solved the structure of ZNF217 bound to its cognate DNA. Conclusion: ZNF217 presents a unique DNA interaction pattern including a new type of protein-DNA contact. Significance: This study deepens our understanding of DNA recognition by classical zinc fingers. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Vandevenne, M. and Jacques, D.A. and Artuz, C. and Nguyen, C.D. and Kwan, A.H.Y. and Segal, D.J. and Matthews, J.M. and Crossley, M. and Guss, J.M. and MacKay, J.P.},
 doi = {10.1074/jbc.M112.441451},
 journal = {Journal of Biological Chemistry},
 number = {15}
}

@article{
 title = {Effects of calcium binding and the hypertrophic cardiomyopathy A8V mutation on the dynamic equilibrium between closed and open conformations of the regulatory N-domain of isolated cardiac troponin C},
 type = {article},
 year = {2013},
 pages = {1950-1962},
 volume = {52},
 id = {a6e23941-429e-3ffe-905a-69c9fa9461ee},
 created = {2023-01-10T01:44:49.760Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:49.760Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Troponin C (TnC) is the calcium-binding subunit of the troponin complex responsible for initiating striated muscle contraction in response to calcium influx. In the skeletal TnC isoform, calcium binding induces a structural change in the regulatory N-domain of TnC that involves a transition from a closed to open structural state and accompanying exposure of a large hydrophobic patch for troponin I (TnI) to subsequently bind. However, little is understood about how calcium primes the N-domain of the cardiac isoform (cTnC) for interaction with the TnI subunit as the open conformation of the regulatory domain of cTnC has been observed only in the presence of bound TnI. Here we use paramagnetic relaxation enhancement (PRE) to characterize the closed to open transition of isolated cTnC in solution, a process that cannot be observed by traditional nuclear magnetic resonance methods. Our PRE data from four spin-labeled monocysteine constructs of isolated cTnC reveal that calcium binding triggers movement of the N-domain helices toward an open state. Fitting of the PRE data to a closed to open transition model reveals the presence of a small population of cTnC molecules in the absence of calcium that possess an open conformation, the level of which increases substantially upon Ca2+ binding. These data support a model in which calcium binding creates a dynamic equilibrium between the closed and open structural states to prime cTnC for interaction with its target peptide. We also used PRE data to assess the structural effects of a familial hypertrophic cardiomyopathy point mutation located within the N-domain of cTnC (A8V). The PRE data show that the Ca2+ switch mechanism is perturbed by the A8V mutation, resulting in a more open N-domain conformation in both the apo and holo states. © 2013 American Chemical Society.},
 bibtype = {article},
 author = {Cordina, N.M. and Liew, C.K. and Gell, D.A. and Fajer, P.G. and MacKay, J.P. and Brown, L.J.},
 doi = {10.1021/bi4000172},
 journal = {Biochemistry},
 number = {11}
}

@article{
 title = {Structural basis of the interaction of the breast cancer Oncogene LMO4 with the tumour suppressor CtIP/RBBP8},
 type = {article},
 year = {2013},
 pages = {1101-1110},
 volume = {425},
 id = {191b6ccd-b3f5-3f12-a76e-f0e33183952f},
 created = {2023-01-10T01:44:50.675Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:50.675Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {LIM-only protein 4 (LMO4) is strongly linked to the progression of breast cancer. Although the mechanisms underlying this phenomenon are not well understood, a role is emerging for LMO4 in regulation of the cell cycle. We determined the solution structure of LMO4 in complex with CtIP (C-terminal binding protein interacting protein)/RBBP8, a tumour suppressor protein that is involved in cell cycle progression, DNA repair and transcriptional regulation. Our data reveal that CtIP and the essential LMO cofactor LDB1 (LIM-domain binding protein 1) bind to the same face on LMO4 and cannot simultaneously bind to LMO4. We hypothesise that overexpression of LMO4 may disrupt some of the normal tumour suppressor activities of CtIP, thereby contributing to breast cancer progression. © 2013 Elsevier Ltd.},
 bibtype = {article},
 author = {Stokes, P.H. and Liew, C.W. and Kwan, A.H. and Foo, P. and Barker, H.E. and Djamirze, A. and O'Reilly, V. and Visvader, J.E. and Mackay, J.P. and Matthews, J.M.},
 doi = {10.1016/j.jmb.2013.01.017},
 journal = {Journal of Molecular Biology},
 number = {7}
}

@article{
 title = {Semiquantitative and quantitative analysis of protein-DNA interactions using steady-state measurements in surface plasmon resonance competition experiments},
 type = {article},
 year = {2013},
 pages = {178-185},
 volume = {440},
 id = {cb78304b-2ce1-32bf-ae2e-25e278ca3b2c},
 created = {2023-01-10T01:44:51.607Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:51.607Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {One method commonly used to characterize protein-DNA interactions is surface plasmon resonance (SPR). In a typical SPR experiment, chip-bound DNA is exposed to increasing concentrations of protein; the resulting binding data are used to calculate a dissociation constant for the interaction. However, in cases in which knowledge of the specificity of the interaction is required, a large set of DNA variants has to be tested; this is time consuming and costly, in part because of the requirement for multiple SPR chips. We have developed a new protocol that uses steady-state binding levels in SPR competition experiments to determine protein-binding dissociation constants for a set of DNA variants. This approach is rapid and straightforward and requires the use of only a single SPR chip. Additionally, in contrast to other methods, our approach does not require prior knowledge of parameters such as on or off rates, using an estimate of the wild-type interaction as the sole input. Utilizing relative steady-state responses, our protocol also allows for the rapid, reliable, and simultaneous determination of protein-binding dissociation constants of a large series of DNA mutants in a single experiment in a semiquantitative fashion. We compare our approach to existing methods, highlighting specific advantages as well as limitations. © 2013 Elsevier Inc. All rights reserved.},
 bibtype = {article},
 author = {Gamsjaeger, R. and Kariawasam, R. and Bang, L.H. and Touma, C. and Nguyen, C.D. and Matthews, J.M. and Cubeddu, L. and Mackay, J.P.},
 doi = {10.1016/j.ab.2013.04.030},
 journal = {Analytical Biochemistry},
 number = {2}
}

@article{
 title = {Analysis of disease-causing GATA1 mutations in murine gene complementation systems},
 type = {article},
 year = {2013},
 pages = {5218-5227},
 volume = {121},
 id = {ddfb0231-026d-32b5-a980-22e8754a9d8b},
 created = {2023-01-10T01:44:52.681Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:52.681Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Missense mutations in transcription factor GATA1 underlie a spectrum of congenital red blood cell and platelet disorders. We investigated how these alterations cause distinct clinical phenotypes by combining structural, biochemical, and genomic approaches with gene complementation systems that examine GATA1 function in biologically relevant cellular contexts. Substitutions that disrupt FOG1 cofactor binding impair both gene activation and repression and are associated with pronounced clinical phenotypes. Moreover, clinical severity correlates with the degree of FOG1 disruption. Surprisingly, 2 mutations shown to impair DNA binding of GATA1 in vitro did not measurably affect in vivo target gene occupancy. Rather, one of these disrupted binding to the TAL1 complex, implicating it in diseases caused by GATA1 mutations. Diminished TAL1 complex recruitment mainly impairs transcriptional activation and is linked to relatively mild disease. Notably, different substitutions at the same amino acid can selectively inhibit TAL1 complex or FOG1 binding, producing distinct cellular and clinical phenotypes. The structure-function relationships elucidated here were not predicted by prior in vitro or computational studies. Thus, our findings uncover novel disease mechanisms underlying GATA1 mutations and highlight the power of gene complementation assays for elucidating the molecular basis of genetic diseases.},
 bibtype = {article},
 author = {Campbell, A.E. and Wilkinson-White, L. and MacKay, J.P. and Matthews, J.M. and Blobel, G.A.},
 doi = {10.1182/blood-2013-03-488080},
 journal = {Blood},
 number = {26}
}

@article{
 title = {Solution structure of the LIM-homeodomain transcription factor complex Lhx3/Ldb1 and the effects of a pituitary mutation on key Lhx3 interactions},
 type = {article},
 year = {2012},
 volume = {7},
 id = {128d3304-f755-382e-92e5-6fc2f2229b4f},
 created = {2023-01-10T01:44:53.740Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:53.740Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Lhx3 is a LIM-homeodomain (LIM-HD) transcription factor that regulates neural cell subtype specification and pituitary development in vertebrates, and mutations in this protein cause combined pituitary hormone deficiency syndrome (CPHDS). The recently published structures of Lhx3 in complex with each of two key protein partners, Isl1 and Ldb1, provide an opportunity to understand the effect of mutations and posttranslational modifications on key protein-protein interactions. Here, we use small-angle X-ray scattering of an Ldb1-Lhx3 complex to confirm that in solution the protein is well represented by our previously determined NMR structure as an ensemble of conformers each comprising two well-defined halves (each made up of LIM domain from Lhx3 and the corresponding binding motif in Ldb1) with some flexibility between the two halves. NMR analysis of an Lhx3 mutant that causes CPHDS, Lhx3(Y114C), shows that the mutation does not alter the zinc-ligation properties of Lhx3, but appears to cause a structural rearrangement of the hydrophobic core of the LIM2 domain of Lhx3 that destabilises the domain and/or reduces the affinity of Lhx3 for both Ldb1 and Isl1. Thus the mutation would affect the formation of Lhx3-containing transcription factor complexes, particularly in the pituitary gland where these complexes are required for the production of multiple pituitary cell types and hormones. © 2012 Bhati et al.},
 bibtype = {article},
 author = {Bhati, M. and Lee, C. and Gadd, M.S. and Jeffries, C.M. and Kwan, A. and Whitten, A.E. and Trewhella, J. and Mackay, J.P. and Matthews, J.M.},
 doi = {10.1371/journal.pone.0040719},
 journal = {PLoS ONE},
 number = {7}
}

@article{
 title = {A rapid method for assessing the RNA-binding potential of a protein},
 type = {article},
 year = {2012},
 volume = {40},
 id = {45db7a3c-9c0b-3ecb-a0b6-8ca4c8c9c8cc},
 created = {2023-01-10T01:44:54.656Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:54.656Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures. © The Author(s) 2012.},
 bibtype = {article},
 author = {Bendak, K. and Loughlin, F.E. and Cheung, V. and O'Connell, M.R. and Crossley, M. and MacKay, J.P.},
 doi = {10.1093/nar/gks285},
 journal = {Nucleic Acids Research},
 number = {14}
}

@article{
 title = {Bivalent recognition of nucleosomes by the tandem PHD fingers of the CHD4 ATPase is required for CHD4-mediated repression},
 type = {article},
 year = {2012},
 pages = {787-792},
 volume = {109},
 id = {fd6d1a67-ad29-356c-9c60-bd33f657e985},
 created = {2023-01-10T01:44:55.575Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:55.575Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {CHD4 is a catalytic subunit of the NuRD (nucleosome remodeling and deacetylase) complex essential in transcriptional regulation, chromatin assembly and DNA damage repair. CHD4 contains tandem plant homeodomain (PHD) fingers connected by a short linker, the biological function of which remains unclear. Here we explore the combinatorial action of the CHD4 PHD1/2 fingers and detail the molecular basis for their association with chromatin. We found that PHD1/2 targets nucleosomes in a multivalent manner, concomitantly engaging two histone H3 tails. This robust synergistic interaction displaces HP1γ from pericentric sites, inducing changes in chromatin structure and leading to the dispersion of the heterochromatic mark H3K9me3. We demonstrate that recognition of the histone H3 tails by the PHD fingers is required for repressive activity of the CHD4/NuRD complex. Together, our data elucidate the molecular mechanism of multivalent association of the PHD fingers with chromatin and reveal their critical role in the regulation of CHD4 functions.},
 bibtype = {article},
 author = {Musselman, C.A. and Ramiŕez, J. and Sims, J.K. and Mansfield, R.E. and Oliver, S.S. and Denu, J.M. and Mackay, J.P. and Wade, P.A. and Hagman, J. and Kutateladze, T.G.},
 doi = {10.1073/pnas.1113655109},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {3}
}

@article{
 title = {Two-timing zinc finger transcription factors liaising with RNA},
 type = {article},
 year = {2012},
 pages = {199-205},
 volume = {37},
 id = {2554db33-7b4b-3127-99cf-3fde4951ed45},
 created = {2023-01-10T01:44:56.496Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:56.496Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Classical zinc fingers (ZFs) are one of the most common protein domains in higher eukaryotes and have been known for almost 30 years to act as sequence-specific DNA-binding domains. This knowledge has come, however, from the study of a small number of archetypal proteins, and a larger picture is beginning to emerge that ZF functions are far more diverse than originally suspected. Here, we review the evidence that a subset of ZF proteins live double lives, binding to both DNA and RNA targets and frequenting both the cytoplasm and the nucleus. This duality can create an important additional level of gene regulation that serves to connect transcriptional and post-transcriptional control. © 2012 Elsevier Ltd.},
 bibtype = {article},
 author = {Burdach, J. and O'Connell, M.R. and Mackay, J.P. and Crossley, M.},
 doi = {10.1016/j.tibs.2012.02.001},
 journal = {Trends in Biochemical Sciences},
 number = {5}
}

@article{
 title = {Solution structure of a tethered Lmo2<inf>LIM2</inf>/Ldb1<inf>LID</inf> complex},
 type = {article},
 year = {2012},
 pages = {1768-1774},
 volume = {21},
 id = {c8d8b2ce-e804-3b1e-bc6d-dea5554342b7},
 created = {2023-01-10T01:44:57.390Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:57.390Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.},
 bibtype = {article},
 author = {Dastmalchi, S. and Wilkinson-White, L. and Kwan, A.H. and Gamsjaeger, R. and Mackay, J.P. and Matthews, J.M.},
 doi = {10.1002/pro.2153},
 journal = {Protein Science},
 number = {11}
}

@article{
 title = {Modular assembly of RanBP2-Type zinc finger domains to target single-stranded RNA},
 type = {article},
 year = {2012},
 pages = {5371-5375},
 volume = {51},
 id = {524d43b0-eac3-303b-9076-d08253f97d3b},
 created = {2023-01-10T01:44:58.301Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:58.301Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Fingering RNA: To study the function of the variety of RNA species that have been discovered recently, sequence-specific RNA-binding molecules with tunable specificity are required. Here, the use of zinc fingers (ZFs) from the splicing factors ZRANB2 and RBM5 as potential modules for the assembly of sequence-specific RNA-binding molecules is demonstrated. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.},
 bibtype = {article},
 author = {O'Connell, M.R. and Vandevenne, M. and Nguyen, C.D. and Matthews, J.M. and Gamsjaeger, R. and Segal, D.J. and MacKay, J.P.},
 doi = {10.1002/anie.201200866},
 journal = {Angewandte Chemie - International Edition},
 number = {22}
}

@article{
 title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular LMO4-LIM1/CtIP complex},
 type = {article},
 year = {2012},
 pages = {31-34},
 volume = {6},
 id = {f121ae58-5c4e-3fbb-987c-35bd8fe224e1},
 created = {2023-01-10T01:44:59.248Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:44:59.248Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {LMO4 is a broadly expressed LIM-only protein that is involved in neural tube development and implicated in breast cancer. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the N-terminal LIM domain from LMO4 tethered to residues 641-685 of C-terminal binding protein interacting protein (CtIP/RBBP8). © 2011 Springer Science+Business Media B.V.},
 bibtype = {article},
 author = {Liew, C.W. and Kwan, A.H. and Stokes, P.H. and MacKay, J.P. and Matthews, J.M.},
 doi = {10.1007/s12104-011-9319-0},
 journal = {Biomolecular NMR Assignments},
 number = {1}
}

@article{
 title = {Synthesis of the bacteriocin glycopeptide sublancin 168 and S-glycosylated variants},
 type = {article},
 year = {2012},
 pages = {1910-1913},
 volume = {14},
 id = {0782aeb9-904a-31dc-bf32-5d68b28b7eb4},
 created = {2023-01-10T01:45:00.240Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:00.240Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The synthesis of sublancin 168, a unique S-glucosylated bacteriocin antibiotic, is described. The natural product and two S-glycosylated variants were successfully prepared via native chemical ligation followed by folding. The synthetic glycopeptides were shown to possess primarily an α-helical secondary structure by CD and NMR studies. © 2012 American Chemical Society.},
 bibtype = {article},
 author = {Hsieh, Y.S.Y. and Wilkinson, B.L. and O'Connell, M.R. and MacKay, J.P. and Matthews, J.M. and Payne, R.J.},
 doi = {10.1021/ol300557g},
 journal = {Organic Letters},
 number = {7}
}

@article{
 title = {Backbone and sidechain <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans},
 type = {article},
 year = {2012},
 pages = {83-86},
 volume = {6},
 id = {ff9a844c-9cab-30ff-b896-a298653e06c7},
 created = {2023-01-10T01:45:01.153Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:01.153Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.},
 bibtype = {article},
 author = {Morris, V.K. and Kwan, A.H. and MacKay, J.P. and Sunde, M.},
 doi = {10.1007/s12104-011-9330-5},
 journal = {Biomolecular NMR Assignments},
 number = {1}
}

@article{
 title = {Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry},
 type = {article},
 year = {2012},
 pages = {1267-1278},
 volume = {18},
 id = {620041f4-f32a-3709-9ae9-50c212e6b3e8},
 created = {2023-01-10T01:45:02.072Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:02.072Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.},
 bibtype = {article},
 author = {Mckenzie, J.L. and Duyvestyn, J.M. and Smith, T. and Bendak, K. and Mackay, J. and Cursons, R.A.Y. and Cook, G.M. and Arcus, V.L.},
 doi = {10.1261/rna.031229.111},
 journal = {RNA},
 number = {6}
}

@article{
 title = {Interdomain orientation of cardiac troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state},
 type = {article},
 year = {2012},
 pages = {1376-1387},
 volume = {21},
 id = {4e767ca7-1e6a-3aae-aa86-c051dd322f85},
 created = {2023-01-10T01:45:03.028Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:03.028Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.},
 bibtype = {article},
 author = {Cordina, N.M. and Liew, C.K. and Gell, D.A. and Fajer, P.G. and Mackay, J.P. and Brown, L.J.},
 doi = {10.1002/pro.2124},
 journal = {Protein Science},
 number = {9}
}

@article{
 title = {Self-assembly of functional, amphipathic amyloid monolayers by the fungal hydrophobin EAS},
 type = {article},
 year = {2012},
 volume = {109},
 id = {b585d9ee-1807-3f19-9891-d5c72141f251},
 created = {2023-01-10T01:45:04.060Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:04.060Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The hydrophobin EAS from the fungus Neurospora crassa forms functional amyloid fibrils called rodlets that facilitate spore formation and dispersal. Self-assembly of EAS into fibrillar rodlets occurs spontaneously at hydrophobic:hydrophilic interfaces and the rodlets further associate laterally to form amphipathic monolayers. We have used site-directed mutagenesis and peptide experiments to identify the region of EAS that drives intermolecular association and formation of the cross-β rodlet structure. Transplanting this region into a nonamyloidogenic hydrophobin enables it to form rodlets. We have also determined the structure and dynamics of an EAS variant with reduced rodlet-forming ability. Taken together, these data allow us to pinpoint the conformational changes that take place when hydrophobins self-assemble at an interface and to propose a model for the amphipathic EAS rodlet structure.},
 bibtype = {article},
 author = {Macindoe, I. and Kwan, A.H. and Ren, Q. and Morris, V.K. and Yang, W. and Mackay, J.P. and Sunde, M.},
 doi = {10.1073/pnas.1114052109},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {14}
}

@book{
 title = {The detection and quantitation of protein oligomerization},
 type = {book},
 year = {2012},
 source = {Advances in Experimental Medicine and Biology},
 pages = {19-41},
 volume = {747},
 id = {7e3e3789-bc90-34ed-805a-f49344268f6e},
 created = {2023-01-10T01:45:04.982Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:04.982Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {There are many different techniques available to biologists and biochemists that can be used to detect and characterize the self-association of proteins. Each technique has strengths and weaknesses and it is often useful to combine several approaches to maximize the former and minimize the latter. Here we review a range of methodologies that identify protein self-association and/or allow the stoichiometry and affinity of the interaction to be determined, placing an emphasis on what type of information can be obtained and outlining the advantages and disadvantages involved. In general, in vitro biophysical techniques, such as size exclusion chromatography, analytical ultracentrifugation, scattering techniques, NMR spectroscopy, isothermal titration calorimetry, fluorescence anisotropy and mass spectrometry, provide information on stoichiometry and/or binding affinities. Other approaches such as cross-linking, fluorescence methods (e.g., fluorescence correlation spectroscopy, FCS; Förster resonance energy transfer, FRET; fluorescence recovery after photobleaching, FRAP; and proximity imaging, PRIM) and complementation approaches (e.g., yeast two hybrid assays and bimolecular fluorescence complementation, BiFC) can be used to detect protein self-association in a cellular context. © 2012 Springer Science+Business Media, LLC.},
 bibtype = {book},
 author = {Gell, D.A. and Grant, R.P. and MacKay, J.P.},
 doi = {10.1007/978-1-4614-3229-6_2}
}

@article{
 title = {The multi-zinc finger protein ZNF217 contacts DNA through a two-finger domain},
 type = {article},
 year = {2011},
 pages = {38190-38201},
 volume = {286},
 id = {e1fdc7f2-dcf3-344a-b417-57036c8d3fcc},
 created = {2023-01-10T01:45:05.935Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:05.935Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Classical C2H2 zinc finger proteins are among the most abundant transcription factors found in eukaryotes, and the mechanisms through which they recognize their target genes have been extensively investigated. In general, a tandem array of three fingers separated by characteristic TGERP links is required for sequence-specific DNA recognition. Nevertheless, a significant number of zinc finger proteins do not contain a hallmark three-finger array of this type, raising the question of whether and how they contact DNA. We have examined the multi-finger protein ZNF217, which contains eight classical zinc fingers. ZNF217 is implicated as an oncogene and in repressing the E-cadherin gene. We show that two of its zinc fingers, 6 and 7, can mediate contacts with DNA. We examine its putative recognition site in the E-cadherin promoter and demonstrate that this is a suboptimal site. NMR analysis and mutagenesis is used to define the DNA binding surface of ZNF217, and we examine the specificity of the DNA binding activity using fluorescence anisotropy titrations. Finally, sequence analysis reveals that a variety of multi-finger proteins also contain two-finger units, and our data support the idea that these may constitute a distinct subclass of DNA recognition motif. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A.},
 bibtype = {article},
 author = {Nunez, N. and Clifton, M.M.K. and Funnell, A.P.W. and Artuz, C. and Hallal, S. and Quinlan, K.G.R. and Font, J. and Vandevenne, M. and Setiyaputra, S. and Pearson, R.C.M. and Mackay, J.P. and Crossley, M.},
 doi = {10.1074/jbc.M111.301234},
 journal = {Journal of Biological Chemistry},
 number = {44}
}

@article{
 title = {Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes},
 type = {article},
 year = {2011},
 volume = {108},
 id = {cd0868d1-a20c-3300-8452-75a3d9552903},
 created = {2023-01-10T01:45:06.903Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:06.903Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 "reads" acetyl marks on nuclear factors to promote their stable association with chromatin.},
 bibtype = {article},
 author = {Lamonica, J.M. and Deng, W. and Kadauke, S. and Campbell, A.E. and Gamsjaeger, R. and Wang, H. and Cheng, Y. and Billin, A.N. and Hardison, R.C. and Mackay, J.P. and Mackay, J.P. and Blobel, G.A.},
 doi = {10.1073/pnas.1102140108},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {22}
}

@article{
 title = {Structural basis for hemoglobin capture by Staphylococcus aureus cell-surface protein, IsdH},
 type = {article},
 year = {2011},
 pages = {38439-38447},
 volume = {286},
 id = {206f34b7-fe09-3a3c-a885-250484075b13},
 created = {2023-01-10T01:45:07.824Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:07.824Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Pathogens must steal iron from their hosts to establish infection. In mammals, hemoglobin (Hb) represents the largest reservoir of iron, and pathogens express Hb-binding proteins to access this source. Here, we show how one of the commonest and most significant human pathogens, Staphylococcus aureus, captures Hb as the first step of an iron-scavenging pathway. The x-ray crystal structure of Hb bound to a domain from the Isd (iron-regulated surface determinant) protein, IsdH, is the first structure of a Hb capture complex to be determined. Surface mutations in Hb that reduce binding to the Hb-receptor limit the capacity of S. aureus to utilize Hb as an iron source, suggesting that Hb sequence is a factor in host susceptibility to infection. The demonstration that pathogens make highly specific recognition complexes with Hb raises the possibility of developing inhibitors of Hb binding as antibacterial agents. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Kumar, K.K. and Jacques, D.A. and Pishchany, G. and Caradoc-Davies, T. and Spirig, T. and Malmirchegini, G.R. and Langley, D.B. and Dickson, C.F. and Mackay, J.P. and Clubb, R.T. and Guss, J.M. and Gell, D.A.},
 doi = {10.1074/jbc.M111.287300},
 journal = {Journal of Biological Chemistry},
 number = {44}
}

@article{
 title = {Protein-protein interactions: Analysis of a false positive GST pulldown result},
 type = {article},
 year = {2011},
 pages = {2365-2371},
 volume = {79},
 id = {5d9c6e9b-c8ce-3c98-ab37-684192c58e66},
 created = {2023-01-10T01:45:08.736Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:08.736Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {One of the most common ways to demonstrate a direct protein-protein interaction in vitro is the glutathione-S-transferse (GST)-pulldown. Here we report the detailed characterization of a putative interaction between two transcription factor proteins, GATA-1 and Krüppel-like factor 3 (KLF3/BKLF) that show robust interactions in GST-pulldown experiments. Attempts to map the interaction interface of GATA-1 on KLF3 using a mutagenic screening approach did not yield a contiguous binding face on KLF3, suggesting that the interaction might be non-specific. NMR experiments showed that the proteins do not interact at protein concentrations of 50-100 μM. Rather, the GST tag can cause part of KLF3 to misfold. In addition to misfolding, the fact that both proteins are DNA-binding domains appears to introduce binding artifacts (possibly nucleic acid bridging) that cannot be resolved by the addition of nucleases or ethidium bromide (EtBr). This study emphasizes the need for caution in relying on GST-pulldown results and related methods, without convincing confirmation from different approaches. © 2011 Wiley-Liss, Inc.},
 bibtype = {article},
 author = {Wissmueller, S. and Font, J. and Liew, C.W. and Cram, E. and Schroeder, T. and Turner, J. and Crossley, M. and Mackay, J.P. and Matthews, J.M.},
 doi = {10.1002/prot.23068},
 journal = {Proteins: Structure, Function and Bioinformatics},
 number = {8}
}

@article{
 title = {Structural basis and specificity of acetylated transcription factor GATA1 recognition by BET family bromodomain protein Brd3},
 type = {article},
 year = {2011},
 pages = {2632-2640},
 volume = {31},
 id = {429de600-d97d-3c2c-9f33-97c9b5577dcd},
 created = {2023-01-10T01:45:09.655Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:09.655Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Recent data demonstrate that small synthetic compounds specifically targeting bromodomain proteins can modulate the expression of cancer-related or inflammatory genes. Although these studies have focused on the ability of bromodomains to recognize acetylated histones, it is increasingly becoming clear that histone-like modifications exist on other important proteins, such as transcription factors. However, our understanding of the molecular mechanisms through which these modifications modulate protein function is far from complete. The transcription factor GATA1 can be acetylated at lysine residues adjacent to the zinc finger domains, and this acetylation is essential for the normal chromatin occupancy of GATA1. We have recently identified the bromodomain-containing protein Brd3 as a cofactor that interacts with acetylated GATA1 and shown that this interaction is essential for the targeting of GATA1 to chromatin. Here we describe the structural basis for this interaction. Our data reveal for the first time the molecular details of an interaction between a transcription factor bearing multiple acetylation modifications and its cognate recognition module. We also show that this interaction can be inhibited by an acetyllysine mimic, highlighting the importance of further increasing the specificity of compounds that target bromodomain and extraterminal (BET) bromodomains in order to fully realize their therapeutic potential. © 2011, American Society for Microbiology.},
 bibtype = {article},
 author = {Gamsjaeger, R. and Webb, S.R. and Lamonica, J.M. and Billin, A. and Blobel, G.A. and Mackay, J.P.},
 doi = {10.1128/MCB.05413-11},
 journal = {Molecular and Cellular Biology},
 number = {13}
}

@article{
 title = {Plant homeodomain (PHD) fingers of CHD4 are histone H3-binding modules with preference for unmodified H3K4 and methylated H3K9},
 type = {article},
 year = {2011},
 pages = {11779-11791},
 volume = {286},
 id = {5cbc3d31-ec2f-35ec-a0c2-85fb87abd270},
 created = {2023-01-10T01:45:10.576Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:10.576Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {A major challenge in chromatin biology is to understand the mechanisms by which chromatin is remodeled into active or inactive states as required during development and cell differentiation. One complex implicated in these processes is the nucleosome remodeling and histone deacetylase (NuRD) complex, which contains both histone deacetylase and nucleosome remodeling activities and has been implicated in the silencing of subsets of genes involved in various stages of cellular development. Chromodomain-helicase-DNA-binding protein 4 (CHD4) is a core component of the NuRD complex and contains a nucleosome remodeling ATPase domain along with two chromodomains and two plant homeodomain (PHD) fingers. We have previously demonstrated that the second PHD finger of CHD4 binds peptides corresponding to the N terminus of histone H3 methylated at Lys9. Here, we determine the solution structure of PHD2 in complex with H3K9me3, revealing the molecular basis of histone recognition, including a cation-π recognition mechanism for methylated Lys9. Additionally, we demonstrate that the first PHD finger also exhibits binding to the N terminus of H3, and we establish the histone-binding surface of this domain. This is the first instance where histone binding ability has been demonstrated for two separate PHD modules within the one protein. These findings suggest that CHD4 could bind to two H3 N-terminal tails on the same nucleosome or on two separate nucleosomes simultaneously, presenting exciting implications for the mechanism by which CHD4 and the NuRD complex could direct chromatin remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Mansfield, R.E. and Musselman, C.A. and Kwan, A.H. and Oliver, S.S. and Garske, A.L. and Davrazou, F. and Denu, J.M. and Kutateladze, T.G. and Mackay, J.P.},
 doi = {10.1074/jbc.M110.208207},
 journal = {Journal of Biological Chemistry},
 number = {13}
}

@article{
 title = {Characterization of a family of RanBP2-Type zinc fingers that can recognize single-stranded RNA},
 type = {article},
 year = {2011},
 pages = {273-283},
 volume = {407},
 id = {48de412f-0ebf-3005-9f78-2ff45c65410e},
 created = {2023-01-10T01:45:11.498Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:11.498Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The recognition of single-stranded RNA (ssRNA) is an important aspect of gene regulation, and a number of different classes of protein domains that recognize ssRNA in a sequence-specific manner have been identified. Recently, we demonstrated that the RanBP2-type zinc finger (ZnF) domains from the human splicing factor ZnF Ran binding domain-containing protein 2 (ZRANB2) can bind to a sequence containing the consensus AGGUAA. Six other human proteins, namely, Ewing's sarcoma (EWS), translocated in liposarcoma (TLS)/FUS, RNA-binding protein 56 (RBP56), RNA-binding motif 5 (RBM5), RNA-binding motif 10 (RBM10) and testis-expressed sequence 13A (TEX13A), each contains a single ZnF with homology to the ZRANB2 ZnFs, and several of these proteins have been implicated in the regulation of mRNA processing. Here, we show that all of these ZnFs are able to bind with micromolar affinities to ssRNA containing a GGU motif. NMR titration data reveal that binding is mediated by the corresponding surfaces on each ZnF, and we also show that sequence selectivity is largely limited to the GGU core motif and that substitution of the three flanking adenines that were selected in our original selection experiment has a minimal effect on binding affinity. These data establish a subset of RanBP2-type ZnFs as a new family of ssRNA-binding motifs. © 2011 Elsevier Ltd.},
 bibtype = {article},
 author = {Nguyen, C.D. and Mansfield, R.E. and Leung, W. and Vaz, P.M. and Loughlin, F.E. and Grant, R.P. and MacKay, J.P.},
 doi = {10.1016/j.jmb.2010.12.041},
 journal = {Journal of Molecular Biology},
 number = {2}
}

@article{
 title = {Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1},
 type = {article},
 year = {2011},
 pages = {14443-14448},
 volume = {108},
 id = {765574bf-c2e8-3a13-8a8c-db663be1661f},
 created = {2023-01-10T01:45:12.413Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:12.413Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (<50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.},
 bibtype = {article},
 author = {Wilkinson-White, L. and Gamsjaeger, R. and Dastmalchi, S. and Wienert, B. and Stokes, P.H. and Crossley, M. and Mackay, J.P. and Matthews, J.M.},
 doi = {10.1073/pnas.1105898108},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {35}
}

@article{
 title = {Macromolecular NMR spectroscopy for the non-spectroscopist: Beyond macromolecular solution structure determination},
 type = {article},
 year = {2011},
 pages = {704-715},
 volume = {278},
 id = {0c69b0a8-cbe5-3ad1-acb3-27b8003b448f},
 created = {2023-01-10T01:45:13.329Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:13.329Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {A strength of NMR spectroscopy is its ability to monitor, on an atomic level, molecular changes and interactions. In this review, which is intended for non-spectroscopist, we describe major uses of NMR in protein science beyond solution structure determination. After first touching on how NMR can be used to quickly determine whether a mutation induces structural perturbations in a protein, we describe the unparalleled ability of NMR to monitor binding interactions over a wide range of affinities, molecular masses and solution conditions. We discuss the use of NMR to measure the dynamics of proteins at the atomic level and over a wide range of timescales. Finally, we outline new and expanding areas such as macromolecular structure determination in multicomponent systems, as well as in the solid state and in vivo. © 2011 FEBS.},
 bibtype = {article},
 author = {Bieri, M. and Kwan, A.H. and Mobli, M. and King, G.F. and MacKay, J.P. and Gooley, P.R.},
 doi = {10.1111/j.1742-4658.2011.08005.x},
 journal = {FEBS Journal},
 number = {5}
}

@article{
 title = {Erratum: The prospects for designer single-stranded RNA-binding proteins (Nature Structural and Molecular Biology (2011) 18 (256-261))},
 type = {article},
 year = {2011},
 pages = {516},
 volume = {18},
 id = {af45e7f3-95df-3e38-b1e4-265877fed464},
 created = {2023-01-10T01:45:14.370Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:14.370Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {MacKay, J.P. and Font, J. and Segal, D.J.},
 doi = {10.1038/nsmb0411-516e},
 journal = {Nature Structural and Molecular Biology},
 number = {4}
}

@article{
 title = {Macromolecular NMR spectroscopy for the non-spectroscopist},
 type = {article},
 year = {2011},
 pages = {687-703},
 volume = {278},
 id = {ed62160d-f565-3f31-ab39-6f55529dd09e},
 created = {2023-01-10T01:45:15.274Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:15.274Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {NMR spectroscopy is a powerful tool for studying the structure, function and dynamics of biological macromolecules. However, non-spectroscopists often find NMR theory daunting and data interpretation nontrivial. As the first of two back-to-back reviews on NMR spectroscopy aimed at non-spectroscopists, the present review first provides an introduction to the basics of macromolecular NMR spectroscopy, including a discussion of typical sample requirements and what information can be obtained from simple NMR experiments. We then review the use of NMR spectroscopy for determining the 3D structures of macromolecules and examine how to judge the quality of NMR-derived structures. © 2011 FEBS.},
 bibtype = {article},
 author = {Kwan, A.H. and Mobli, M. and Gooley, P.R. and King, G.F. and MacKay, J.P.},
 doi = {10.1111/j.1742-4658.2011.08004.x},
 journal = {FEBS Journal},
 number = {5}
}

@article{
 title = {Insights into association of the NuRD complex with FOG-1 from the crystal structure of an RbAp48·FOG-1 complex},
 type = {article},
 year = {2011},
 pages = {1196-1203},
 volume = {286},
 id = {0dcda68f-44aa-33d1-afea-ef433768b02c},
 created = {2023-01-10T01:45:16.197Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:16.197Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Chromatin-modifying complexes such as the NuRD complex are recruited to particular genomic sites by gene-specific nuclear factors. Overall, however, little is known about the molecular basis for these interactions. Here, we present the 1.9 Å resolution crystal structure of the NuRD subunit RbAp48 bound to the 15 N-terminal amino acids of the GATA-1 cofactor FOG-1. The FOG-1 peptide contacts a negatively charged binding pocket on top of the RbAp48 β-propeller that is distinct from the binding surface used by RpAp48 to contact histone H4. We further show that RbAp48 interacts with the NuRD subunit MTA-1 via a surface that is distinct from its FOG-binding pocket, providing a first glimpse into the way in which NuRD assembly facilitates interactions with cofactors. Our RbAp48·FOG-1 structure provides insight into the molecular determinants of FOG-1-dependent association with the NuRD complex and into the links between transcription regulation and nucleosome remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Lejon, S. and Thong, S.Y. and Murthy, A. and AlQarni, S. and Murzina, N.V. and Blobel, G.A. and Laue, E.D. and Mackay, J.P.},
 doi = {10.1074/jbc.M110.195842},
 journal = {Journal of Biological Chemistry},
 number = {2}
}

@article{
 title = {The prospects for designer single-stranded RNA-binding proteins},
 type = {article},
 year = {2011},
 pages = {256-261},
 volume = {18},
 id = {ff45d397-50a9-36fc-8a33-31782437940d},
 created = {2023-01-10T01:45:17.102Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:17.102Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Spectacular progress has been made in the design of proteins that recognize double-stranded DNA with a chosen specificity, to the point that designer DNA-binding proteins can be ordered commercially. This success raises the question of whether it will be possible to engineer libraries of proteins that can recognize RNA with tailored specificity. Given the recent explosion in the number and diversity of RNA species demonstrated to play roles in biology, designer RNA-binding proteins are set to become valuable tools, both in the research laboratory and potentially in the clinic. Here we discuss the prospects for the realization of this idea. © 2011 Nature America, Inc. All rights reserved.},
 bibtype = {article},
 author = {MacKay, J.P. and Font, J. and Segal, D.J.},
 doi = {10.1038/nsmb.2005},
 journal = {Nature Structural and Molecular Biology},
 number = {3}
}

@article{
 title = {The structure of a truncated phosphoribosylanthranilate isomerase suggests a unified model for evolution of the (βα)<inf>8</inf> barrel fold},
 type = {article},
 year = {2011},
 pages = {291-303},
 volume = {408},
 id = {484786a4-2a38-33ce-8896-5cba612e7ef8},
 created = {2023-01-10T01:45:18.013Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:18.013Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The (βα)8 barrel is one of the most common protein folds, and enzymes with this architecture display a remarkable range of catalytic activities. Many of these functions are associated with ancient metabolic pathways, and phylogenetic reconstructions suggest that the (βα)8 barrel was one of the very first protein folds to emerge. Consequently, there is considerable interest in understanding the evolutionary processes that gave rise to this fold. In particular, much attention has been focused on the plausibility of (βα)8 barrel evolution from homodimers of half barrels. However, we previously isolated a three-quarter-barrel-sized fragment of a (βα)8 barrel, termed truncated phosphoribosylanthranilate isomerase (trPRAI), that is soluble and almost as thermostable as full-length N-(5′-phosphoribosyl) anthranilate isomerase (PRAI). Here, we report the NMR-derived structure of trPRAI. The subdomain is monomeric, is well ordered and adopts a native-like structure in solution. Side chains from strands β1 (Glu3 and Lys5), β2 (Tyr25) and β6 (Lys122) of trPRAI repack to shield the hydrophobic core from the solvent. This result demonstrates that three-quarter barrels were viable intermediates in the evolution of the (βα)8 barrel fold. We propose a unified model for (βα)8 barrel evolution that combines our data, previously published work and plausible scenarios for the emergence of (initially error-prone) genetic systems. In this model, the earliest proto-cells contained diverse pools of part-barrel subdomains. Combinatorial assembly of these subdomains gave rise to many distinct lineages of (βα)8 barrel proteins, that is, our model excludes the possibility that there was a single (βα)8 barrel from which all present examples are descended. © 2011 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Setiyaputra, S. and MacKay, J.P. and Patrick, W.M.},
 doi = {10.1016/j.jmb.2011.02.048},
 journal = {Journal of Molecular Biology},
 number = {2}
}

@article{
 title = {AHSP (α-haemoglobin-stabilizing protein) stabilizes apo-α-haemoglobin in a partially folded state},
 type = {article},
 year = {2010},
 pages = {275-282},
 volume = {432},
 id = {cfeb53fe-7617-3568-b0ac-944abaedb834},
 created = {2023-01-10T01:45:18.928Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:18.928Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {To produce functional Hb (haemoglobin), nascent α-globin (αo) and β-globin (βo) chains must each bind a single haem molecule (to form αh and βh) and interact together to form heterodimers.The precise sequence of binding events is unknown, and it has been suggested that additional factors might enhance the efficiency of Hb folding. AHSP (α-haemoglobin-stabilizing protein) has been shown previously to bind αh and regulate redox activity of the haem iron. In the present study, we used a combination of classical and dynamic light scattering and NMR spectroscopy to demonstrate that AHSP forms a heterodimeric complex with αo that inhibits αo aggregation and promotes αo folding in the absence of haem. These findings indicate that AHSP may function as an αo-specific chaperone, and suggest an important role for αo in guiding Hb assembly by stabilizing βo and inhibiting off-pathway self-association of βh. © The Authors Journal compilation © 2010 Biochemical Society.},
 bibtype = {article},
 author = {Krishna Kumar, K. and Dickson, C.F. and Weiss, M.J. and Mackay, J.P. and Gell, D.A.},
 doi = {10.1042/BJ20100642},
 journal = {Biochemical Journal},
 number = {2}
}

@book{
 title = {Beyond DNA: Zinc finger domains as RNA-binding modules},
 type = {book},
 year = {2010},
 source = {Methods in Molecular Biology},
 pages = {479-491},
 volume = {649},
 id = {c6c74829-cc59-3c5a-bd21-95b1c21c0aac},
 created = {2023-01-10T01:45:19.842Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:19.842Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.},
 bibtype = {book},
 author = {Font, J. and MacKay, J.P.},
 doi = {10.1007/978-1-60761-753-2_29}
}

@book{
 title = {Preface},
 type = {book},
 year = {2010},
 source = {Methods in Molecular Biology},
 volume = {649},
 id = {7ce2996b-3608-389e-b725-b249c989c5b4},
 created = {2023-01-10T01:45:20.744Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:20.744Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {book},
 author = {MacKay, J.P. and Segal, D.J.}
}

@article{
 title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular Lmo2-LIM2/Ldb1-LID complex},
 type = {article},
 year = {2010},
 pages = {203-206},
 volume = {4},
 id = {d3067811-3bab-3f57-99ac-522a48b4c9b3},
 created = {2023-01-10T01:45:21.656Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:21.656Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Lmo2 is a LIM-only protein involved in hematopoiesis and the development of T-cell acute lymphoblastic leukaemia. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the C-terminal LIM domain from Lmo2 tethered to the LIM interaction domain (LID) from LIM domain binding protein 1 (Ldb1). © 2010 Springer Science+Business Media B.V.},
 bibtype = {article},
 author = {Wilkinson-White, L.E. and Dastmalchi, S. and Kwan, A.H. and Ryan, D.P. and MacKay, J.P. and Matthews, J.M.},
 doi = {10.1007/s12104-010-9240-y},
 journal = {Biomolecular NMR Assignments},
 number = {2}
}

@article{
 title = {Two-state conformational equilibrium in the Par-4 leucine zipper domain},
 type = {article},
 year = {2010},
 pages = {2433-2449},
 volume = {78},
 id = {f7152d3f-03a8-3aab-8417-8bbf484d2488},
 created = {2023-01-10T01:45:22.571Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:22.571Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.},
 bibtype = {article},
 author = {Schwalbe, M. and Dutta, K. and Libich, D.S. and Venugopal, H. and Claridge, J.K. and Gell, D.A. and Mackay, J.P. and Edwards, P.J.B. and Pascal, S.M.},
 doi = {10.1002/prot.22752},
 journal = {Proteins: Structure, Function and Bioinformatics},
 number = {11}
}

@article{
 title = {<sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis},
 type = {article},
 year = {2010},
 pages = {167-169},
 volume = {4},
 id = {032a0fa0-83ef-3610-bdcc-94e56a23bb75},
 created = {2023-01-10T01:45:23.480Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:23.480Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.},
 bibtype = {article},
 author = {Hynson, R.M.G. and Kwan, A.H. and Jacques, D.A. and MacKay, J.P. and Trewhella, J.},
 doi = {10.1007/s12104-010-9237-6},
 journal = {Biomolecular NMR Assignments},
 number = {2}
}

@book{
 title = {Protein Recognition},
 type = {book},
 year = {2010},
 source = {Amino Acids, Peptides and Proteins in Organic Chemistry},
 pages = {505-532},
 volume = {2},
 id = {cf609a26-bd7d-39ca-a12c-7a19ab224d44},
 created = {2023-01-10T01:45:24.389Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:24.389Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {book},
 author = {Mansfield, R.E. and Cross, A.J. and Matthews, J.M. and Mackay, J.P.},
 doi = {10.1002/9783527631780.ch12}
}

@article{
 title = {The structural analysis of protein-protein interactions by NMR spectroscopy},
 type = {article},
 year = {2009},
 pages = {5224-5232},
 volume = {9},
 id = {388e096a-15bd-345c-96eb-0a1702c4a9b6},
 created = {2023-01-10T01:45:25.437Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:25.437Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.},
 bibtype = {article},
 author = {O'Connell, M.R. and Gamsjaeger, R. and Mackay, J.P.},
 doi = {10.1002/pmic.200900303},
 journal = {Proteomics},
 number = {23}
}

@article{
 title = {Conformational Stability and DNA Binding Specificity of the Cardiac T-Box Transcription Factor Tbx20},
 type = {article},
 year = {2009},
 pages = {606-618},
 volume = {389},
 id = {c62d0b72-ffae-3a09-8c2e-63a5222cc5a2},
 created = {2023-01-10T01:45:26.344Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:26.344Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The transcription factor Tbx20 acts within a hierarchy of T-box factors in lineage specification and morphogenesis in the mammalian heart and is mutated in congenital heart disease. T-box family members share a ∼ 20-kDa DNA-binding domain termed the T-box. The question of how highly homologous T-box proteins achieve differential transcriptional control in heart development, while apparently binding to the same DNA sequence, remains unresolved. Here we show that the optimal DNA recognition sequence for the T-box of Tbx20 corresponds to a T-half-site. Furthermore, we demonstrate using purified recombinant domains that distinct T-boxes show significant differences in the affinity and kinetics of binding and in conformational stability, with the T-box of Tbx20 displaying molten globule character. Our data highlight unique features of Tbx20 and suggest mechanistic ways in which cardiac T-box factors might interact synergistically and/or competitively within the cardiac regulatory network. © 2009 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Macindoe, I. and Glockner, L. and Vukašin, P. and Stennard, F.A. and Costa, M.W. and Harvey, R.P. and Mackay, J.P. and Sunde, M.},
 doi = {10.1016/j.jmb.2009.04.056},
 journal = {Journal of Molecular Biology},
 number = {3}
}

@article{
 title = {A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin},
 type = {article},
 year = {2009},
 pages = {29462-29469},
 volume = {284},
 id = {a280cdd1-9f7b-37f1-8e93-e668c33b7b99},
 created = {2023-01-10T01:45:27.256Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:27.256Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {α-Hemoglobin (αHb) stabilizing protein (AHSP) is expressed in erythropoietic tissues as an accessory factor in hemoglobin synthesis. AHSP forms a specific complex with αHb and suppresses the heme-catalyzed evolution of reactive oxygen species by converting αHb to a conformation in which the heme is coordinated at both axial positions by histidine side chains (bis-histidyl coordination). Currently, the detailed mechanism by which AHSP induces structural changes in αHb has not been determined. Here, we present x-ray crystallography, NMR spectroscopy, and mutagenesis data that identify, for the first time, the importance of an evolutionarily conserved proline, Pro30, in loop 1 of AHSP. Mutation of Pro30 to a variety of residue types results in reduced ability to convert αHb. In complex with-Hb, AHSP Pro30 adopts a cis-peptidyl conformation and makes contact with the N terminus of helix G in αHb. Mutations that stabilize the cis-peptidyl conformation of free AHSP, also enhance the αHb conversion activity. These findings suggest that AHSP loop 1 can transmit structural changes to the heme pocket of αHb, and, more generally, highlight the importance of cis-peptidyl prolyl residues in defining the conformation of regulatory protein loops. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Gell, D.A. and Feng, L. and Zhou, S. and Jeffrey, P.D. and Bendak, K. and Gow, A. and Weiss, M.J. and Shi, Y. and Mackay, J.P.},
 doi = {10.1074/jbc.M109.027045},
 journal = {Journal of Biological Chemistry},
 number = {43}
}

@article{
 title = {Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications},
 type = {article},
 year = {2009},
 pages = {179-187},
 volume = {423},
 id = {852e680d-f3c3-3677-a4d0-54c4e5f30958},
 created = {2023-01-10T01:45:28.180Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:28.180Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation ormethylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2-H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin. © The Authors Journal compilation. © 2009 Biochemical Society.},
 bibtype = {article},
 author = {Musselman, C.A. and Mansfield, R.E. and Garske, A.L. and Davrazou, F. and Kwan, A.H. and Oliver, S.S. and O'Leary, H. and Denu, J.M. and Mackay, J.P. and Kutateladze, T.G.},
 doi = {10.1042/BJ20090870},
 journal = {Biochemical Journal},
 number = {2}
}

@article{
 title = {Erratum: Structure and inhibition of orotidine 5#-Monophosphate decarboxylase from plasmodium falciparum (Biochemistry (2009) 48:11 (2570-2570) 10.1021/bi900043p)},
 type = {article},
 year = {2009},
 pages = {2570},
 volume = {48},
 id = {600819d9-be33-3029-9db3-d7575a665191},
 created = {2023-01-10T01:45:29.095Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:29.095Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Langley, D.B. and Shojaei, M. and Chan, C. and Lok, H.C. and Mackay, J.P. and Traut, T.W. and Guss, J.M. and Christopherson, R.I.},
 doi = {10.1021/bi900043p},
 journal = {Biochemistry},
 number = {11}
}

@article{
 title = {A mass spectrometric investigation of the ability of metal complexes to modulate transcription factor activity},
 type = {article},
 year = {2009},
 pages = {5546-5548},
 id = {674f52e4-6615-311d-9893-ba1bd7e3946c},
 created = {2023-01-10T01:45:30.041Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:30.041Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {ESI mass spectrometry was used to assess the ability of metal complexes to inhibit binding of a transcription factor to a DNA molecule containing its recognition sequence. © 2009 The Royal Society of Chemistry.},
 bibtype = {article},
 author = {Talib, J. and Beck, J.L. and Urathamakul, T. and Nguyen, C.D. and Aldrich-Wright, J.R. and MacKay, J.P. and Ralph, S.F.},
 doi = {10.1039/b904751d},
 journal = {Chemical Communications},
 number = {37}
}

@article{
 title = {Structural analysis of MED-1 reveals unexpected diversity in the mechanism of DNA recognition by GATA-type zinc finger domains},
 type = {article},
 year = {2009},
 pages = {5827-5835},
 volume = {284},
 id = {71df5cc8-308b-387c-9226-824ae9beba6f},
 created = {2023-01-10T01:45:30.973Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:30.973Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {MED-1 is a member of a group of divergent GATA-type zinc finger proteins recently identified in several species of Caenorhabditis. The med genes are transcriptional regulators that are involved in the specification of the mesoderm and endoderm precursor cells in nematodes. Unlike other GATA-type zinc fingers that recognize the consensus sequence (A/C/ T)GATA(A/G), the MED-1 zinc finger (MED1zf) binds the larger and atypical site GTATACT(T/C)3. We have examined the basis for this unusual DNA specificity using a range of biochemical and biophysical approaches. Most strikingly, we show that although the core of the MED1zf structure is similar to that of GATA-1, the basic tail C-terminal to the zinc finger unexpectedly adopts an α-helical structure upon binding DNA. This additional helix appears to contact the major groove of the DNA, making contacts that explain the extended DNA consensus sequence observed for MED1zf. Our data expand the versatility of DNA recognition by GATA-type zinc fingers and perhaps shed new light on the DNA-binding properties of mammalian GATA factors. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Lowry, J.A. and Gamsjaeger, R. and Thong, S.Y. and Hung, W. and Kwan, A.H. and Broitman-Maduro, G. and Matthews, J.M. and Maduro, M. and Mackay, J.P.},
 doi = {10.1074/jbc.M808712200},
 journal = {Journal of Biological Chemistry},
 number = {9}
}

@article{
 title = {The zinc fingers of the SR-like protein ZRANB2 are single-stranded RNA-binding domains that recognize 5' splice site-like sequences},
 type = {article},
 year = {2009},
 pages = {5581-5586},
 volume = {106},
 id = {bbb64689-b326-3416-95e2-8b6b88f2d5e6},
 created = {2023-01-10T01:45:31.914Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:31.914Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The alternative splicing of mRNA is a critical process in higher eukaryotes that generates substantial proteomic diversity. Many of the proteins that are essential to this process contain arginine/serine-rich (RS) domains. ZRANB2 is a widely-expressed and highly-conserved RS-domain protein that can regulate alternative splicing but lacks canonical RNA-binding domains. Instead, it contains 2 RanBP2-type zinc finger (ZnF) domains. We demonstrate that these ZnFs recognize ssRNA with high affinity and specificity. Each ZnF binds to a single AGGUAA motif and the 2 domains combine to recognize AGGUAA (Nx) AGGUAA double sites, suggesting that ZRANB2 regulates alternative splicing via a direct interaction with pre-mRNA at sites that resemble the consensus 5' splice site. We show using X-ray crystallography that recognition of an AGGUAA motif by a single ZnF is dominated by side-chain hydrogen bonds to the bases and formation of a guanine-tryptophan-guanine ''ladder.'' A number of other human proteins that function in RNA processing also contain RanBP2 ZnFs in which the RNA-binding residues of ZRANB2 are conserved. The ZnFs of ZRANB2 therefore define another class of RNA-binding domain, advancing our understanding of RNA recognition and emphasizing the versatility of domains in molecular recognition.},
 bibtype = {article},
 author = {Loughlin, F.E. and Mansfield, R.E. and Vaz, P.M. and McGrath, A.P. and Setiyaputra, S. and Gamsjaeger, R. and Chen, E.S. and Morris, B.J. and Guss, J.M. and Mackay, J.P.},
 doi = {10.1073/pnas.0802466106},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {14}
}

@article{
 title = {It takes two to tango: The structure and function of LIM, RING, PHD and MYND domains},
 type = {article},
 year = {2009},
 pages = {3681-3696},
 volume = {15},
 id = {12e1371d-f8f7-3ef4-8a33-189b9e2b23cc},
 created = {2023-01-10T01:45:32.880Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:32.880Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {LIM (Lin-11, Isl-1, Mec-3), RING (Really interesting new gene), PHD (Plant homology domain) and MYND (myeloid, Nervy, DEAF-1) domains are all zinc-binding domains that ligate two zinc ions. Unlike the better known classical zinc fingers, these domains do not bind DNA, but instead mediate interactions with other proteins. LIM-domain containing proteins have diverse functions as regulators of gene expression, cell adhesion and motility and signal transduction. RING finger proteins are generally associated with ubiquitination; the presence of such a domain is the defining feature of a class of E3 ubiquitin protein ligases. PHD proteins have been associated with SUMOylation but most recently have emerged as a chromatin recognition motif that reads the methylation state of histones. The function of the MYND domain is less clear, but MYND domains are also found in proteins that have ubiquitin ligase and/or histone methyltransferase activity. Here we review the structure-function relationships for these domains and discuss strategies to modulate their activity. © 2009 Bentham Science Publishers Ltd.},
 bibtype = {article},
 author = {Matthews, J.M. and Bhati, M. and Lehtomaki, E. and Mansfield, R.E. and Cubeddu, L. and Mackay, J.P.},
 doi = {10.2174/138161209789271861},
 journal = {Current Pharmaceutical Design},
 number = {31}
}

@article{
 title = {NMR spectroscopy as a tool for the rapid assessment of the conformation of GST-fusion proteins},
 type = {article},
 year = {2008},
 pages = {1630-1635},
 volume = {17},
 id = {2555f698-f625-3490-b864-b58850f3abab},
 created = {2023-01-10T01:45:33.906Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:33.906Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Glutathione-S-transferase (GST)-fusion proteins are used extensively for structural, biochemical, and functional analyses. Although the conformation of the target protein is of critical importance, confirmation of the folded state of the target is often not undertaken or is cumbersome because of the requirement to first remove the GST tag. Here, we demonstrate that it is possible to record conventional 15N-HSQC NMR spectra of small GST-fusion proteins and that the observed signals arise almost exclusively from the target protein. This approach constitutes a rapid and straightforward means of assessing the conformation of a GST-fusion protein without having to cleave the GST and should prove valuable, both to biochemists seeking to check the conformation of their proteins prior to functional studies and to structural biologists screening protein constructs for suitability as targets for structural studies. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 The Protein Society.},
 bibtype = {article},
 author = {Chu, K.L. and Gamsjaeger, R. and Mansfield, R.E. and Mackay, J.P.},
 doi = {10.1110/ps.034983.108},
 journal = {Protein Science},
 number = {9}
}

@article{
 title = {Structural analysis of hydrophobins},
 type = {article},
 year = {2008},
 pages = {773-784},
 volume = {39},
 id = {d6614c6a-3917-3fb1-bfca-b4688d016cf4},
 created = {2023-01-10T01:45:34.809Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:34.809Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Hydrophobins are a remarkable class of small cysteine-rich proteins found exclusively in fungi. They self-assemble to form robust polymeric monolayers that are highly amphipathic and play numerous roles in fungal biology, such as in the formation and dispersal of aerial spores and in pathogenic and mutualistic interactions. The polymeric form can be reversibly disassembled and is able to reverse the wettability of a surface, leading to many proposals for nanotechnological applications over recent years. The surprising properties of hydrophobins and their potential for commercialization have led to substantial efforts to delineate their morphology and molecular structure. In this review, we summarize the progress that has been made using a variety of spectroscopic and microscopic approaches towards understanding the molecular mechanisms underlying hydrophobin structure. © 2007 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Sunde, M. and Kwan, A.H.Y. and Templeton, M.D. and Beever, R.E. and Mackay, J.P.},
 doi = {10.1016/j.micron.2007.08.003},
 journal = {Micron},
 number = {7}
}

@article{
 title = {The Cys3-Cys4 Loop of the Hydrophobin EAS Is Not Required for Rodlet Formation and Surface Activity},
 type = {article},
 year = {2008},
 pages = {708-720},
 volume = {382},
 id = {209bc497-4431-3395-a923-3ba1135b8fac},
 created = {2023-01-10T01:45:35.837Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:35.837Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Class I hydrophobins are fungal proteins that self-assemble into robust amphipathic rodlet monolayers on the surface of aerial structures such as spores and fruiting bodies. These layers share many structural characteristics with amyloid fibrils and belong to the growing family of functional amyloid-like materials produced by microorganisms. Although the three-dimensional structure of the soluble monomeric form of a class I hydrophobin has been determined, little is known about the molecular structure of the rodlets or their assembly mechanism. Several models have been proposed, some of which suggest that the Cys3-Cys4 loop has a critical role in the initiation of assembly or in the polymeric structure. In order to provide insight into the relationship between hydrophobin sequence and rodlet assembly, we investigated the role of the Cys3-Cys4 loop in EAS, a class I hydrophobin from Neurospora crassa. Remarkably, deletion of up to 15 residues from this 25-residue loop does not impair rodlet formation or reduce the surface activity of the protein, and the physicochemical properties of rodlets formed by this mutant are indistinguishable from those of its full-length counterpart. In addition, the core structure of the truncation mutant is essentially unchanged. Molecular dynamics simulations carried out on the full-length protein and this truncation mutant binding to an air-water interface show that, although it is hydrophobic, the loop does not play a role in positioning the protein at the surface. These results demonstrate that the Cys3-Cys4 loop does not have an integral role in the formation or structure of the rodlets and that the major determinant of the unique properties of these proteins is the amphipathic core structure, which is likely to be preserved in all hydrophobins despite the high degree of sequence variation across the family. © 2008 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Kwan, A.H. and Macindoe, I. and Vukašin, P.V. and Morris, V.K. and Kass, I. and Gupte, R. and Mark, A.E. and Templeton, M.D. and Mackay, J.P. and Sunde, M.},
 doi = {10.1016/j.jmb.2008.07.034},
 journal = {Journal of Molecular Biology},
 number = {3}
}

@article{
 title = {Response to Chatr-aryamontri et al.: Protein interactions: to believe or not to believe?},
 type = {article},
 year = {2008},
 pages = {242-243},
 volume = {33},
 id = {a93b20e9-5f78-368e-9bc7-126a1f8e9c4e},
 created = {2023-01-10T01:45:36.806Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:36.806Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Mackay, J.P. and Sunde, M. and Lowry, J.A. and Crossley, M. and Matthews, J.M.},
 doi = {10.1016/j.tibs.2008.04.003},
 journal = {Trends in Biochemical Sciences},
 number = {6}
}

@article{
 title = {Evolution of Quaternary Structure in a Homotetrameric Enzyme},
 type = {article},
 year = {2008},
 pages = {691-703},
 volume = {380},
 id = {63b6023f-2683-3fd8-a96d-6ee13c0b2a51},
 created = {2023-01-10T01:45:37.717Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:37.717Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Dihydrodipicolinate synthase (DHDPS) is an essential enzyme in (S)-lysine biosynthesis and an important antibiotic target. All X-ray crystal structures solved to date reveal a homotetrameric enzyme. In order to explore the role of this quaternary structure, dimeric variants of Escherichia coli DHDPS were engineered and their properties were compared to those of the wild-type tetrameric form. X-ray crystallography reveals that the active site is not disturbed when the quaternary structure is disrupted. However, the activity of the dimeric enzymes in solution is substantially reduced, and a tetrahedral adduct of a substrate analogue is observed to be trapped at the active site in the crystal form. Remarkably, heating the dimeric enzymes increases activity. We propose that the homotetrameric structure of DHDPS reduces dynamic fluctuations present in the dimeric forms and increases specificity for the first substrate, pyruvate. By restricting motion in a key catalytic motif, a competing, non-productive reaction with a substrate analogue is avoided. Small-angle X-ray scattering and mutagenesis data, together with a B-factor analysis of the crystal structures, support this hypothesis and lead to the suggestion that in at least some cases, the evolution of quaternary enzyme structures might serve to optimise the dynamic properties of the protein subunits. © 2008 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Griffin, M.D.W. and Dobson, R.C.J. and Pearce, F.G. and Antonio, L. and Whitten, A.E. and Liew, C.K. and Mackay, J.P. and Trewhella, J. and Jameson, G.B. and Perugini, M.A. and Perugini, M.A. and Gerrard, J.A.},
 doi = {10.1016/j.jmb.2008.05.038},
 journal = {Journal of Molecular Biology},
 number = {4}
}

@article{
 title = {Implementing the LIM code: The structural basis for cell type-specific assembly of LIM-homeodomain complexes},
 type = {article},
 year = {2008},
 pages = {2018-2029},
 volume = {27},
 id = {aab1c421-98b0-388e-a789-d78011488b4f},
 created = {2023-01-10T01:45:38.632Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:38.632Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {LIM-homeodomain (LIM-HD) transcription factors form a combinatorial 'LIM code' that contributes to the specification of cell types. In the ventral spinal cord, the binary LIM homeobox protein 3 (Lhx3)/LIM domain-binding protein 1 (Ldb1) complex specifies the formation of V2 interneurons. The additional expression of islet-1 (Isl1) in adjacent cells instead specifies the formation of motor neurons through assembly of a ternary complex in which Isl1 contacts both Lhx3 and Ldb1, displacing Lhx3 as the binding partner of Ldb1. However, little is known about how this molecular switch occurs. Here, we have identified the 30-residue Lhx3-binding domain on Isl1 (Isl1LBD). Although the LIM interaction domain of Ldb1 (Ldb1LID) and Isl1LBD share low levels of sequence homology, X-ray and NMR structures reveal that they bind Lhx3 in an identical manner, that is, Isl1LBD mimics Ldb1 LID. These data provide a structural basis for the formation of cell type-specific protein-protein interactions in which unstructured linear motifs with diverse sequences compete to bind protein partners. The resulting alternate protein complexes can target different genes to regulate key biological events. ©2008 European Molecular Biology Organization.},
 bibtype = {article},
 author = {Bhati, M. and Lee, C. and Nancarrow, A.L. and Lee, M. and Craig, V.J. and Bach, I. and Guss, J.M. and Mackay, J.P. and Matthews, J.M.},
 doi = {10.1038/emboj.2008.123},
 journal = {EMBO Journal},
 number = {14}
}

@article{
 title = {Structural and biophysical analysis of the DNA binding properties of myelin transcription factor 1},
 type = {article},
 year = {2008},
 pages = {5158-5167},
 volume = {283},
 id = {915fd038-e9b1-36a4-8be4-dcecb9f56909},
 created = {2023-01-10T01:45:39.551Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:39.551Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Zinc binding domains, or zinc fingers (ZnFs), form one of the most numerous and most diverse superclasses of protein structural motifs in eukaryotes. Although our understanding of the functions of several classes of these domains is relatively well developed, we know much less about the molecular mechanisms of action of many others. Myelin transcription factor 1 (MyT1) type ZnFs are found in organisms as diverse as nematodes and mammals and are found in a range of sequence contexts. MyT1, one of the early transcription factors expressed in the developing central nervous system, contains seven MyT1 ZnFs that are very highly conserved both within the protein and between species. We have used a range of biophysical techniques, including NMR spectroscopy and data-driven macromolecular docking, to investigate the structural basis for the interaction between MyT1 ZnFs and DNA. Our data indicate that MyT1 ZnFs recognize the major groove of DNA in a way that appears to differ from other known zinc binding domains. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Gamsjaeger, R. and Swanton, M.K. and Kobus, F.J. and Lehtomaki, E. and Lowry, J.A. and Kwan, A.H. and Matthews, J.M. and Mackay, J.P.},
 doi = {10.1074/jbc.M703772200},
 journal = {Journal of Biological Chemistry},
 number = {8}
}

@article{
 title = {Structure and inhibition of orotidine 5′-monophosphate decarboxylase from Plasmodium falciparum},
 type = {article},
 year = {2008},
 pages = {3842-3854},
 volume = {47},
 id = {a141dd55-ab8c-3d60-ae66-5bb20bb77e87},
 created = {2023-01-10T01:45:40.465Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:40.465Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Orotidine 5′-monophosphate (OMP) decarboxylase from Plasmodium falciparum (PfODCase, EC 4.1.1.23) has been overexpressed, purified, subjected to kinetic and biochemical analysis, and crystallized. The native enzyme is a homodimer with a subunit molecular mass of 38 kDa. The saturation curve for OMP as a substrate conformed to Michaelis-Menten kinetics with Km = 350 ± 60 nM and Vmax = 2.70 ± 0.10 μmol/min/mg protein. Inhibition patterns for nucleoside 5′-monophosphate analogues were linear competitive with respect to OMP with a decreasing potency of inhibition of PfODCase in the order: pyrazofurin 5′-monophosphate (Ki = 3.6 ± 0.7 nM) > xanthosine 5′-monophosphate (XMP, Ki = 4.4 ± 0.7 nM) > 6-azauridine 5′-monophosphate (AzaUMP, K i = 12 ±3 nM) > allopurinol-3-riboside 5′- monophosphate (Ki = 240 ± 20 nM). XMP is an ∼150-fold more potent inhibitor of PfODCase compared with the human enzyme. The structure of PfODCase was solved in the absence of ligand and displays a classic TIM-barrel fold characteristic of the enzyme. Both the phosphate-binding loop and the βα5-loop have conformational flexibility, which may be associated with substrate capture and product release along the reaction pathway. © 2008 American Chemical Society.},
 bibtype = {article},
 author = {Langley, D.B. and Shojaei, M. and Chan, C. and Hiu, C.L. and Mackay, J.P. and Traut, T.W. and Guss, J.M. and Christopherson, R.I.},
 doi = {10.1021/bi702390k},
 journal = {Biochemistry},
 number = {12}
}

@article{
 title = {Crystallization of a ZRANB2-RNA complex},
 type = {article},
 year = {2008},
 pages = {1175-1177},
 volume = {64},
 id = {5c489013-8140-39aa-85b6-66fd1e85cec0},
 created = {2023-01-10T01:45:41.371Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:41.371Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.},
 bibtype = {article},
 author = {Loughlin, F.E. and Lee, M. and Guss, J.M. and Mackay, J.P.},
 doi = {10.1107/S1744309108036993},
 journal = {Acta Crystallographica Section F: Structural Biology and Crystallization Communications},
 number = {12}
}

@article{
 title = {Designed metal-binding sites in biomolecular and bioinorganic interactions},
 type = {article},
 year = {2008},
 pages = {484-490},
 volume = {18},
 id = {719ca520-9316-3d73-84b3-1e92163b1c32},
 created = {2023-01-10T01:45:42.283Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:42.283Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The design of metal-binding functionality in proteins is expanding into many different areas with a wide range of practical and research applications. Here we review several developing areas of metal-related protein design, including the use of metals to induce protein-protein interactions or facilitate the assembly of extended nanostructures; the design of metallopeptides that bind metal and other inorganic surfaces, an area with potential in diverse applications ranging from nanoelectronics and photonics to biotechnology and biomedicine; and, the creation of sensitive and selective metal sensors for use both in vivo and in vitro. © 2008 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Matthews, J.M. and Loughlin, F.E. and Mackay, J.P.},
 doi = {10.1016/j.sbi.2008.04.009},
 journal = {Current Opinion in Structural Biology},
 number = {4}
}

@article{
 title = {Escherichia coli glucuronylsynthase: An engineered enzyme for the synthesis of β-glucuronides},
 type = {article},
 year = {2008},
 pages = {1585-1588},
 volume = {10},
 id = {d014882a-497b-3d62-96a2-9f6d9e1e973d},
 created = {2023-01-10T01:45:43.190Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:43.190Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The glycosynthase derived from E. coli β-glucuronidase catalyzes the glucuronylation of a range of primary, secondary, and aryl alcohols with moderate to excellent yields. The procedure provides an efficient, stereoselective, and scalable single-step synthesis of β-glucuronides under mild conditions. © 2008 American Chemical Society.},
 bibtype = {article},
 author = {Wilkinson, S.M. and Liew, C.W. and Mackay, J.P. and Salleh, H.M. and Withers, S.G. and McLeod, M.D.},
 doi = {10.1021/ol8002767},
 journal = {Organic Letters},
 number = {8}
}

@article{
 title = {Grb7 SH2 domain structure and interactions with a cyclic peptide inhibitor of cancer cell migration and proliferation},
 type = {article},
 year = {2007},
 volume = {7},
 id = {4beae229-a038-3821-9cc9-5285dcb8776b},
 created = {2023-01-10T01:45:44.136Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:44.136Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background. Human growth factor receptor bound protein 7 (Grb7) is an adapter protein that mediates the coupling of tyrosine kinases with their downstream signaling pathways. Grb7 is frequently overexpressed in invasive and metastatic human cancers and is implicated in cancer progression via its interaction with the ErbB2 receptor and focal adhesion kinase (FAK) that play critical roles in cell proliferation and migration. It is thus a prime target for the development of novel anti-cancer therapies. Recently, an inhibitory peptide (G7-18NATE) has been developed which binds specifically to the Grb7 SH2 domain and is able to attenuate cancer cell proliferation and migration in various cancer cell lines. Results. As a first step towards understanding how Grb7 may be inhibited by G7-18NATE, we solved the crystal structure of the Grb7 SH2 domain to 2.1 Å resolution. We describe the details of the peptide binding site underlying target specificity, as well as the dimer interface of Grb 7 SH2. Dimer formation of Grb7 was determined to be in the μM range using analytical ultracentrifugation for both full-length Grb7 and the SH2 domain alone, suggesting the SH2 domain forms the basis of a physiological dimer. ITC measurements of the interaction of the G7-18NATE peptide with the Grb7 SH2 domain revealed that it binds with a binding affinity of Kd= ∼35.7 μM and NMR spectroscopy titration experiments revealed that peptide binding causes perturbations to both the ligand binding surface of the Grb7 SH2 domain as well as to the dimer interface, suggesting that dimerisation of Grb7 is impacted on by peptide binding. Conclusion. Together the data allow us to propose a model of the Grb7 SH2 domain/G7-18NATE interaction and to rationalize the basis for the observed binding specificity and affinity. We propose that the current study will assist with the development of second generation Grb7 SH2 domain inhibitors, potentially leading to novel inhibitors of cancer cell migration and invasion. © 2007 Porter et al; licensee BioMed Central Ltd.},
 bibtype = {article},
 author = {Porter, C.J. and Matthews, J.M. and Mackay, J.P. and Pursglove, S.E. and Schmidberger, J.W. and Leedman, P.J. and Pero, S.C. and Krag, D.N. and Wilce, M.C.J. and Wilce, J.A.},
 doi = {10.1186/1472-6807-7-58},
 journal = {BMC Structural Biology}
}

@article{
 title = {Sticky fingers: zinc-fingers as protein-recognition motifs},
 type = {article},
 year = {2007},
 pages = {63-70},
 volume = {32},
 id = {0bdbbb8b-8f2a-39d3-a9c6-8cef76dd27e8},
 created = {2023-01-10T01:45:45.060Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:45.060Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Zinc-fingers (ZnFs) are extremely abundant in higher eukaryotes. Once considered to function exclusively as sequence-specific DNA-binding motifs, ZnFs are now known to have additional activities such as the recognition of RNA and other proteins. Here we discuss recent advances in our understanding of ZnFs as specific modules for protein recognition. Structural studies of ZnF complexes reveal considerable diversity in terms of protein partners, binding modes and affinities, and highlight the often underestimated versatility of ZnF structure and function. An appreciation of the structural features of ZnF-protein interactions will contribute to our ability to engineer and to use ZnFs with tailored protein-binding properties. © 2006 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Gamsjaeger, R. and Liew, C.K. and Loughlin, F.E. and Crossley, M. and Mackay, J.P.},
 doi = {10.1016/j.tibs.2006.12.007},
 journal = {Trends in Biochemical Sciences},
 number = {2}
}

@article{
 title = {Solution Structure of the THAP Domain from Caenorhabditis elegans C-terminal Binding Protein (CtBP)},
 type = {article},
 year = {2007},
 pages = {382-390},
 volume = {366},
 id = {5e95220d-3a32-3190-836d-ac33617c7af2},
 created = {2023-01-10T01:45:45.962Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:45.962Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The THAP (Thanatos-associated protein) domain is a recently discovered zinc-binding domain found in proteins involved in transcriptional regulation, cell-cycle control, apoptosis and chromatin modification. It contains a single zinc atom ligated by cysteine and histidine residues within a Cys-X2-4-Cys-X35-53-Cys-X2-His consensus. We have determined the NMR solution structure of the THAP domain from Caenorhabditis elegans C-terminal binding protein (CtBP) and show that it adopts a fold containing a treble clef motif, bearing similarity to the zinc finger-associated domain (ZAD) from Drosophila Grauzone. The CtBP THAP domain contains a large, positively charged surface patch and we demonstrate that this domain can bind to double-stranded DNA in an electrophoretic mobility-shift assay. These data, together with existing reports, indicate that THAP domains might exhibit a functional diversity similar to that observed for classical and GATA-type zinc fingers. © 2006 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Liew, C.K. and Crossley, M. and Mackay, J.P. and Nicholas, H.R.},
 doi = {10.1016/j.jmb.2006.11.058},
 journal = {Journal of Molecular Biology},
 number = {2}
}

@article{
 title = {An erythroid chaperone that facilitates folding of α-globin subunits for hemoglobin synthesis},
 type = {article},
 year = {2007},
 pages = {1856-1865},
 volume = {117},
 id = {b36c185f-d2dd-356f-b1e4-9fa2eef4f751},
 created = {2023-01-10T01:45:46.881Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:46.881Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Erythrocyte precursors produce abundant α- and β-globin proteins, which assemble with each other to form hemoglobin A (HbA), the major blood oxygen carrier. αHb-stabilizing protein (AHSP) binds free α subunits reversibly to maintain their structure and limit their ability to generate reactive oxygen species. Accordingly, loss of AHSP aggravates the toxicity of excessive free α-globin caused by β-globin gene disruption in mice. Surprisingly, we found that AHSP also has important functions when free α-globin is limited. Thus, compound mutants lacking both Ahsp and 1 of 4 α-globin genes (genotype Ahsp-/-α-globin -/-α/αα) exhibited more severe anemia and Hb instability than mice with either mutation alone. In vitro, recombinant AHSP promoted folding of newly translated α-globin, enhanced its refolding after denaturation, and facilitated its incorporation into HbA. Moreover, in erythroid precursors, newly formed free α-globin was destabilized by loss of AHSP. Therefore, in addition to its previously defined role in detoxification of excess α-globin, AHSP also acts as a molecular chaperone to stabilize nascent α-globin for HbA assembly. Our findings illustrate what we believe to be a novel adaptive mechanism by which a specialized cell coordinates high-level production of a multisubunit protein and protects against various synthetic imbalances.},
 bibtype = {article},
 author = {Yu, X. and Kong, Y. and Dore, L.C. and Abdulmalik, O. and Katein, A.M. and Zhou, S. and Choi, J.K. and Gell, D. and Mackay, J.P. and Gow, A.J. and Gow, A.J. and Weiss, M.J.},
 doi = {10.1172/JCI31664},
 journal = {Journal of Clinical Investigation},
 number = {7}
}

@article{
 title = {Mutations in crdiac T-box factor gene TBX20 are associated with diverse cardiac pathologies, including defects of septation and valvulogenesis and cardiomyopathy},
 type = {article},
 year = {2007},
 pages = {280-291},
 volume = {81},
 id = {52c04b2f-8782-380e-959a-24222788591c},
 created = {2023-01-10T01:45:47.818Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:47.818Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The T-box family transcription factor gene TBX20 acts in a conserved regulatory network, guiding heart formation and patterning in diverse species. Mouse Tbx20 is expressed in cardiac progenitor cells, differentiating cardiomyocytes, and developing valvular tissue, and its deletion or RNA interference-mediated knockdown is catastrophic for heart development. TBX20 interacts physically, functionally, and genetically with other cardiac transcription factors, including NKX2-5, GATA4, and TBX5, mutations of which cause congenital heart disease (CHD). Here, we report nonsense (Q195X) and missense (I152M) germline mutations within the T-box DNA-binding domain of human TBX20 that were associated with a family history of CHD and a complex spectrum of developmental anomalies, including defects in septation, chamber growth, and valvulogenesis. Biophysical characterization of wild-type and mutant proteins indicated how the missense mutation disrupts the structure and function of the TBX20 T-box. Dilated cardiomyopathy was a feature of the TBX20 mutant phenotype in humans and mice, suggesting that mutations in developmental transcription factors can provide a sensitized template for adult-onset heart disease. Our findings are the first to link TBX20 mutations to human pathology. They provide insights into how mutation of different genes in an interactive regulatory circuit lead to diverse clinical phenotypes, with implications for diagnosis, genetic screening, and patient follow-up. © 2007 by The American Society of Human Genetics. All rights reserved.},
 bibtype = {article},
 author = {Kirk, E.P. and Sunde, M. and Costa, M.W. and Rankin, S.A. and Wolstein, O. and Castro, M.L. and Butler, T.L. and Hyun, C. and Guo, G. and Otway, R. and Winlaw, D.S. and Harvey, R.P.},
 doi = {10.1086/519530},
 journal = {American Journal of Human Genetics},
 number = {2}
}

@article{
 title = {Protein interactions: is seeing believing?},
 type = {article},
 year = {2007},
 pages = {530-531},
 volume = {32},
 id = {44b87550-332a-36df-aa40-16703ffad894},
 created = {2023-01-10T01:45:48.739Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:48.739Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Mackay, J.P. and Sunde, M. and Lowry, J.A. and Crossley, M. and Matthews, J.M.},
 doi = {10.1016/j.tibs.2007.09.006},
 journal = {Trends in Biochemical Sciences},
 number = {12}
}

@article{
 title = {Crystal structures of flax rust avirulence proteins AvrL567-A and -D reveal details of the structural basis for flax disease resistance specificity},
 type = {article},
 year = {2007},
 pages = {2898-2912},
 volume = {19},
 id = {e42e696d-217e-3568-9613-ef60ec204073},
 created = {2023-01-10T01:45:49.665Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:49.665Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The gene-for-gene mechanism of plant disease resistance involves direct or indirect recognition of pathogen avirulence (Avr) proteins by plant resistance (R) proteins. Flax rust (Melampsora lini) AvrL567 avirulence proteins and the corresponding flax (Linum usitatissimum) L5, L6, and L7 resistance proteins interact directly. We determined the three-dimensional structures of two members of the AvrL567 family, AvrL567-A and AvrL567-D, at 1.4- and 2.3-Å resolution, respectively. The structures of both proteins are very similar and reveal a β-sandwich fold with no close known structural homologs. The polymorphic residues in the AvrL567 family map to the surface of the protein, and polymorphisms in residues associated with recognition differences for the R proteins lead to significant changes in surface chemical properties. Analysis of single amino acid substitutions in AvrL567 proteins confirm the role of individual residues in conferring differences in recognition and suggest that the specificity results from the cumulative effects of multiple amino acid contacts. The structures also provide insights into possible pathogen-associated functions of AvrL567 proteins, with nucleic acid binding activity demonstrated in vitro. Our studies provide some of the first structural information on avirulence proteins that bind directly to the corresponding resistance proteins, allowing an examination of the molecular basis of the interaction with the resistance proteins as a step toward designing new resistance specificities. © 2007 American Society of Plant Biologists.},
 bibtype = {article},
 author = {Wang, C.-I.A. and Gunčar, G. and Forwood, J.K. and Teh, T. and Catanzariti, A.-M. and Lawrence, G.J. and Loughlin, F.E. and Mackay, J.P. and Schirra, H.J. and Anderson, P.A. and Dodds, P.N. and Kobe, B.},
 doi = {10.1105/tpc.107.053611},
 journal = {Plant Cell},
 number = {9}
}

@article{
 title = {Zinc fingers are known as domains for binding DNA and RNA. Do they also mediate protein-protein interactions?},
 type = {article},
 year = {2006},
 pages = {731-733},
 volume = {58},
 id = {a59a3ace-fadf-3a89-ab6c-424f0813e6d6},
 created = {2023-01-10T01:45:50.577Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:50.577Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Loughlin, F.E. and Mackay, J.P.},
 doi = {10.1080/15216540600868445},
 journal = {IUBMB Life},
 number = {12}
}

@article{
 title = {Molecular analysis of the interaction between the hematopoietic master transcription factors GATA-1 and PU},
 type = {article},
 year = {2006},
 pages = {28296-28306},
 volume = {281},
 id = {62d88558-80bb-327f-9a17-f07a26210a6c},
 created = {2023-01-10T01:45:51.530Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:51.530Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {GATA-1 and PU.1 are transcription factors that control erythroid and myeloid development, respectively. The two proteins have been shown to function in an antagonistic fashion, with GATA-1 repressing PU.1 activity during erythropoiesis and PU.1 repressing GATA-1 function during myelopoiesis. It has also become clear that this functional antagonism involves direct interactions between the two proteins. However, the molecular basis for these interactions is not known, and a number of inconsistencies exist in the literature. We have used a range of biophysical methods to define the molecular details of the GATA-1-PU.1 interaction. A combination of NMR titration data and extensive mutagenesis revealed that the PU.1-Ets domain and the GATA-1 C-terminal zinc finger (CF) form a low affinity interaction in which specific regions of each protein are implicated. Surprisingly, the interaction cannot be disrupted by single alanine substitution mutations, suggesting that binding is distributed over an extended interface. The C-terminal basic tail region of CF appears to be sufficient to mediate an interaction with PU.1-Ets, and neither acetylation nor phosphorylation of a peptide corresponding to this region disrupts binding, indicating that the interaction is not dominated by electrostatic interactions. The CF basic tail shares significant sequence homology with the PU.1 interacting motif from c-Jun, suggesting that GATA-1 and c-Jun might compete to bind PU.1. Taken together, our data provide a molecular perspective on the GATA-1-PU.1 interaction, resolving several issues in the existing data and providing insight into the mechanisms through which these two proteins combine to regulate blood development. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Chu, W.L. and Rand, K.D. and Simpson, R.J.Y. and Yung, W.W. and Mansfield, R.E. and Crossley, M. and Prœtorius-Ibba, M. and Nerlov, C. and Poulsen, F.M. and Mackay, J.P.},
 doi = {10.1074/jbc.M602830200},
 journal = {Journal of Biological Chemistry},
 number = {38}
}

@article{
 title = {Identification of the Key LMO2-binding Determinants on Ldb1},
 type = {article},
 year = {2006},
 pages = {66-75},
 volume = {359},
 id = {14513112-4f7f-308b-b9ed-cb4aa5fd5f5b},
 created = {2023-01-10T01:45:52.515Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:52.515Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The overexpression of LIM-only protein 2 (LMO2) in T-cells, as a result of chromosomal translocations, retroviral insertion during gene therapy, or in transgenic mice models, leads to the onset of T-cell leukemias. LMO2 comprises two protein-binding LIM domains that allow LMO2 to interact with multiple protein partners, including LIM domain-binding protein 1 (Ldb1, also known as CLIM2 and NLI), an essential cofactor for LMO proteins. Sequestration of Ldb1 by LMO2 in T-cells may prevent it binding other key partners, such as LMO4. Here, we show using protein engineering and enzyme-linked immunosorbent assay (ELISA) methodologies that LMO2 binds Ldb1 with a twofold lower affinity than does LMO4. Thus, excess LMO2 rather than an intrinsically higher binding affinity would lead to sequestration of Ldb1. Both LIM domains of LMO2 are required for high-affinity binding to Ldb1 (KD=2.0×10-8 M). However, the first LIM domain of LMO2 is primarily responsible for binding to Ldb1 (KD=2.3×10-7 M), whereas the second LIM domain increases binding by an order of magnitude. We used mutagenesis in combination with yeast two-hybrid analysis, and phage display selection to identify LMO2-binding "hot spots" within Ldb1 that locate to the LIM1-binding region. The delineation of this region reveals some specific differences when compared to the equivalent LMO4:Ldb1 interaction that hold promise for the development of reagents to specifically bind LMO2 in the treatment of leukemia. © 2006 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Ryan, D.P. and Sunde, M. and Kwan, A.H.-Y. and Marianayagam, N.J. and Nancarrow, A.L. and vanden Hoven, R.N. and Thompson, L.S. and Baca, M. and Mackay, J.P. and Visvader, J.E. and Visvader, J.E. and Matthews, J.M.},
 doi = {10.1016/j.jmb.2006.02.074},
 journal = {Journal of Molecular Biology},
 number = {1}
}

@article{
 title = {Structural basis for rodlet assembly in fungal hydrophobins},
 type = {article},
 year = {2006},
 pages = {3621-3626},
 volume = {103},
 id = {0944c3ce-33ab-33ae-b13f-43193a7e6f03},
 created = {2023-01-10T01:45:53.429Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:53.429Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.},
 bibtype = {article},
 author = {Kwan, A.H.Y. and Winefield, R.D. and Sunde, M. and Matthews, J.M. and Haverkamp, R.G. and Templeton, M.D. and Mackay, J.P.},
 doi = {10.1073/pnas.0505704103},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {10}
}

@article{
 title = {Analysis of the structure and function of the transcriptional coregulator HOP},
 type = {article},
 year = {2006},
 pages = {10584-10590},
 volume = {45},
 id = {ad62d749-5828-3220-9e44-df8440515254},
 created = {2023-01-10T01:45:54.347Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:54.347Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Homeodomain-only protein (HOP) is an 8-kDa transcriptional corepressor that is essential for the normal development of the mammalian heart. Previous studies have shown that HOP, which consists entirely of a putative homeodomain, acts downstream of Nkx2.5 and associates with the serum response factor (SRF), repressing transcription from SRF-responsive genes. HOP is also able to recruit histone deacetylase (HDAC) activity, consistent with its ability to repress transcription. Unlike other classic homeodomain proteins, HOP does not appear to interact with DNA, although it has been unclear if this is because of an overall divergent structure or because of specific amino acid differences between HOP and other homeodomains. To work toward an understanding of HOP function, we have determined the 3D structure of full-length HOP and used a range of biochemical assays to define the parts of the protein that are functionally important for its repression activity. We show that HOP forms a classical homeodomain fold but that it cannot recognize double stranded DNA, a result that emphasizes the importance of caution in predicting protein function from sequence homology alone. We also demonstrate that two distinct regions on the surface of HOP are required for its ability to repress an SRF-driven reporter gene, and it is likely that these motifs direct interactions between HOP and partner proteins such as SRF- and HDAC-containing complexes. Our results demonstrate that the homeodomain fold has been co-opted during evolution for functions other than sequence-specific DNA binding and suggest that HOP functions as an adaptor protein to mediate transcriptional repression. © 2006 American Chemical Society.},
 bibtype = {article},
 author = {Kook, H. and Yung, W.W. and Simpson, R.J. and Kee, H.J. and Shin, S. and Lowry, J.A. and Loughlin, F.E. and Yin, Z. and Epstein, J.A. and Mackay, J.P.},
 doi = {10.1021/bi060641s},
 journal = {Biochemistry},
 number = {35}
}

@article{
 title = {A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP},
 type = {article},
 year = {2006},
 pages = {287-297},
 volume = {362},
 id = {f66fd98b-e46d-356c-adc5-dbb7bb45a1c1},
 created = {2023-01-10T01:45:55.259Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:55.259Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Gell, D.A. and Westman, B.J. and Gorman, D. and Liew, C. and Welch, J.J. and Weiss, M.J. and Mackay, J.P.},
 doi = {10.1016/j.jmb.2006.07.010},
 journal = {Journal of Molecular Biology},
 number = {2}
}

@article{
 title = {GATA-1: One protein, many partners},
 type = {article},
 year = {2006},
 pages = {6-11},
 volume = {38},
 id = {91c99222-cb02-3632-8751-7f2b89a9578c},
 created = {2023-01-10T01:45:56.163Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:56.163Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Lowry, J.A. and MacKay, J.P.},
 doi = {10.1016/j.biocel.2005.06.017},
 journal = {International Journal of Biochemistry and Cell Biology},
 number = {1}
}

@article{
 title = {Grb7-SH2 domain dimerisation is affected by a single point mutation},
 type = {article},
 year = {2005},
 pages = {454-460},
 volume = {34},
 id = {273a7a2d-ac31-3984-b4ee-f7adf13fb08c},
 created = {2023-01-10T01:45:57.071Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:57.071Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 μM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation. © EBSA 2005.},
 bibtype = {article},
 author = {Porter, C.J. and Wilce, M.C.J. and Mackay, J.P. and Leedman, P. and Wilce, J.A.},
 doi = {10.1007/s00249-005-0480-1},
 journal = {European Biophysics Journal},
 number = {5}
}

@article{
 title = {Structure of oxidized α-haemoglobin bound to AHSP reveals a protective mechanism for haem},
 type = {article},
 year = {2005},
 pages = {697-701},
 volume = {435},
 id = {a2006ec4-0c0d-3521-8be7-b22dddf9fea3},
 created = {2023-01-10T01:45:57.993Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:57.993Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The synthesis of haemoglobin A (HbA) is exquisitely coordinated during erythrocyte development to prevent damaging effects from individual α- and β-subunits. The α-haemoglobin-stabilizing protein (AHSP) binds α-haemoglobin (αHb), inhibits the ability of αHb to generate reactive oxygen species and prevents its precipitation on exposure to oxidant stress. The structure of AHSP bound to ferrous αHb is thought to represent a transitional complex through which αHb is converted to a non-reactive, hexacoordinate ferric form. Here we report the crystal structure of this ferric αHb-AHSP complex at 2.4 A resolution. Our findings reveal a striking bis-histidyl configuration in which both the proximal and the distal histidines coordinate the haem iron atom. To attain this unusual conformation, segments of αHb undergo drastic structural rearrangements, including the repositioning of several α-helices. Moreover, conversion to the ferric bishistidine configuration strongly and specifically inhibits redox chemistry catalysis and haem loss from αHb. The observed structural changes, which impair the chemical reactivity of haem iron, explain how AHSP stabilizes αHb and prevents its damaging effects in cells.},
 bibtype = {article},
 author = {Feng, L. and Zhou, S. and Gu, L. and Gell, D.A. and Mackay, J.P. and Weiss, M.J. and Gow, A.J. and Shi, Y.},
 doi = {10.1038/nature03609},
 journal = {Nature},
 number = {7042}
}

@article{
 title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular Lhx3:ldb1 complex [4]},
 type = {article},
 year = {2005},
 pages = {198},
 volume = {33},
 id = {f9aa56cf-83b9-3151-a514-a77d7bddc008},
 created = {2023-01-10T01:45:58.923Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:58.923Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Lee, C. and Nancarrow, A.L. and Bach, I. and Mackay, J.P. and Matthews, J.M.},
 doi = {10.1007/s10858-005-3209-7},
 journal = {Journal of Biomolecular NMR},
 number = {3}
}

@article{
 title = {Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/friend of GATA interaction},
 type = {article},
 year = {2005},
 pages = {583-588},
 volume = {102},
 id = {6ca8effa-53df-38c4-a21a-ded042ec2309},
 created = {2023-01-10T01:45:59.921Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:45:59.921Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.},
 bibtype = {article},
 author = {Liew, C.K. and Simpson, R.J.Y. and Kwan, A.H.Y. and Crofts, L.A. and Loughlin, F.E. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},
 doi = {10.1073/pnas.0407511102},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {3}
}

@article{
 title = {Assessment of the robustness of a serendipitous zinc binding fold: Mutagenesis and protein grafting},
 type = {article},
 year = {2005},
 pages = {257-266},
 volume = {13},
 id = {9aeeb5d0-0dee-3dab-93d2-266c720ce66c},
 created = {2023-01-10T01:46:00.845Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:00.845Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Zinc binding motifs have received much attention in the area of protein design. Here, we have tested the suitability of a recently discovered nonnative zinc binding structure as a protein design scaffold. A series of multiple alanine mutants was created to investigate the minimal requirements for folding, and solution structures of these mutants showed that the original fold was maintained, despite changes in 50% of the sequence. We next attempted to transplant binding faces from chosen bimolecular interactions onto one of these mutants, and many of the resulting "chimeras" were shown to adopt a native-like fold. These results both highlight the robust nature of small zinc binding domains and underscore the complexity of designing functional proteins, even using such small, highly ordered scaffolds as templates.},
 bibtype = {article},
 author = {Sharpe, B.K. and Liew, C.K. and Kwan, A.H. and Wilce, J.A. and Crossley, M. and Matthews, J.M. and Mackay, J.P.},
 doi = {10.1016/j.str.2004.12.007},
 journal = {Structure},
 number = {2}
}

@article{
 title = {Solution structure of a recombinant type I sculpin antifreeze protein},
 type = {article},
 year = {2005},
 pages = {1980-1988},
 volume = {44},
 id = {02c3451d-5f79-3362-9ed9-2542509ffb84},
 created = {2023-01-10T01:46:01.762Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:01.762Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {We have determined the solution structure of rSS3, a recombinant form of the type I shorthorn sculpin antifreeze protein (AFP), at 278 and 268 K. This AFP contains an unusual sequence of N-terminal residues, together with two of the 11-residue repeats that are characteristic of the type I winter flounder AFP. The solution conformation of the N-terminal region of the sculpin AFP has been assumed to be the critical factor that results in recognition of different ice planes by the sculpin and flounder AFPs. At 278 K, the two repeats units (residues 11-20 and 21-32) in rSS3 form a continuous α-helix, with the residues 30-33 in the second repeat somewhat less well defined. Within the N-terminal region, residues 2-6 are well defined and helical and linked to the main helix by a more flexible region comprising residues A7-T11. At 268 K the AFP is overall more helical but retains the apparent hinge region. The helical conformation of the two repeats units is almost identical to the corresponding repeats in the type I winter flounder AFP. We also show that while tetracetylated rSS3 has antifreeze activity comparable to the natural AFP, its overall structure is the same as that of the unacetylated peptide. These data provide some insight into the structural determinants of antifreeze activity and should assist in the development of models that explain the recognition of different ice interfaces by the sculpin and flounder type I AFPs.},
 bibtype = {article},
 author = {Kwan, A.H.-Y. and Fairley, K. and Anderberg, P.I. and Liew, C.W. and Harding, M.M. and Mackay, J.P.},
 doi = {10.1021/bi047782j},
 journal = {Biochemistry},
 number = {6}
}

@book{
 title = {Role of alpha hemoglobin-stabilizing protein in normal erythropoiesis and β-thalassemia},
 type = {book},
 year = {2005},
 source = {Annals of the New York Academy of Sciences},
 pages = {103-117},
 volume = {1054},
 id = {3eadb043-bae4-3de9-9a30-64811004299d},
 created = {2023-01-10T01:46:02.725Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:02.725Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Hemoglobin (Hb) synthesis is coordinated by homeostatic mechanisms to limit the accumulation of free α or β subunits, which are cytotoxic. Alpha hemoglobin-stabilizing protein (AHSP) is an abundant erythroid protein that specifically binds free αHb, stabilizes its structure, and limits its ability to participate in chemical reactions that generate reactive oxygen species. Gene ablation studies in mice demonstrate that AHSP is required for normal erythropoiesis. AHSP-null erythrocytes are short-lived, contain Hb precipitates, and exhibit signs of oxidative damage. Loss of AHSP exacerbates β-thalassemia in mice, indicating that altered AHSP expression or function could modify thalassemia phenotypes in humans, a topic that is beginning to be explored in clinical studies. We used biochemical, spectroscopic, and crystallographic methods to examine how AHSP stabilizes αHb. AHSP binds the G and H helices of αHb on a surface that largely overlaps with the α1-β1 interface of HbA. This result explains previous findings that βHb can competitively displace AHSP from αHb to form HbA tetramer. Remarkably, binding of AHSP to oxygenated αHb induces dramatic conformational changes and converts the heme-bound iron to an oxidized hemichrome state in which all six coordinate positions are occupied. This structure limits the reactivity of heme iron, providing a mechanism by which AHSP stabilizes αHb. These findings suggest a biochemical pathway through which AHSP might participate in normal Hb synthesis and modulate the severity of thalassemias. Moreover, understanding how AHSP stabilizes αHb provides a theoretical basis for new strategies to inhibit the damaging effects of free αHb that accumulates in β-thalassemia. © 2005 New York Academy of Sciences.},
 bibtype = {book},
 author = {Weiss, M.J. and Zhou, S. and Feng, L. and Gell, D.A. and Mackay, J.P. and Shi, Y. and Gow, A.J.},
 doi = {10.1196/annals.1345.013}
}

@article{
 title = {The hydrophobic domain 26 of human tropoelastin is unstructured in solution},
 type = {article},
 year = {2005},
 pages = {154-162},
 volume = {150},
 id = {3e2d6e2e-f905-3566-81be-3cc4ef02fb5c},
 created = {2023-01-10T01:46:03.625Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:03.625Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Elastin is the protein responsible for the elastic properties of vertebrate tissue. Very little is currently known about the structure of elastin or of its soluble precursor tropoelastin. We have used high-resolution solution NMR methods to probe the conformational preferences of a conserved hydrophobic region in tropoelastin, domain 26 (D26). Using a combination of homonuclear, 15N-separated and triple resonance experiments, we have obtained essentially full chemical shift assignments for D26 at 278 K. An analysis of secondary chemical shift changes, as well as NOE and 15N relaxation data, leads us to conclude that this domain is essentially unstructured in solution and does not interact with intact tropoelastin. D26 does not display exposed hydrophobic clusters, as expected for a fully unfolded protein and commensurate with an absence of flexible structural motifs, as identified by lack of binding of the fluorescent probe 4,4′-dianilino-1,1′- binaphthyl-5,5′-disulfonic acid. Sedimentation equilibrium data establish that this domain is strictly monomeric in solution. NMR spectra recorded at 278 and 308 K indicate that no significant structural changes occur for this domain over the temperature range 278-308 K, in contrast to the characteristic coacervation behavior that is observed for the full-length protein. © 2005 Elsevier Inc. All rights reserved.},
 bibtype = {article},
 author = {MacKay, J.P. and Muiznieks, L.D. and Toonkool, P. and Weiss, A.S.},
 doi = {10.1016/j.jsb.2005.02.005},
 journal = {Journal of Structural Biology},
 number = {2}
}

@article{
 title = {A complex mechanism determines polarity of DNA replication fork arrest by the replication terminator complex of Bacillus subtilis},
 type = {article},
 year = {2005},
 pages = {13105-13113},
 volume = {280},
 id = {826a04da-c81b-3dcb-bff3-7b7d2e7fede1},
 created = {2023-01-10T01:46:04.549Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:04.549Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Two dimers of the replication terminator protein (RTP) of Bacillus subtilis bind to a chromosomal DNA terminator site to effect polar replication fork arrest. Cooperative binding of the dimers to overlapping half-sites within the terminator is essential for arrest. It was suggested previously that polarity of fork arrest is the result of the RTP dimer at the blocking (proximal) side within the complex binding very tightly and the permissive-side RTP dimer binding relatively weakly. In order to investigate this "differential binding affinity" model, we have constructed a series of mutant terminators that contain half-sites of widely different RTP binding affinities in various combinations. Although there appeared to be a correlation between binding affinity at the proximal half-site and fork arrest efficiency in vivo for some terminators, several deviated significantly from this correlation. Some terminators exhibited greatly reduced binding cooperativity (and therefore have reduced affinity at each half-site) but were highly efficient in fork arrest, whereas one terminator had normal affinity over the proximal half-site, yet had low fork arrest efficiency. The results show clearly that there is no direct correlation between the RTP binding affinity (either within the full complex or at the proximal half-site within the full complex) and the efficiency of replication fork arrest in vivo. Thus, the differential binding affinity over the proximal and distal half-sites cannot be solely responsible for functional polarity of fork arrest. Furthermore, efficient fork arrest relies on features in addition to the tight binding of RTP to terminator DNA. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc.},
 bibtype = {article},
 author = {Duggin, I.G. and Matthews, J.M. and Dixon, N.E. and Wake, R.G. and Mackay, J.P.},
 doi = {10.1074/jbc.M414187200},
 journal = {Journal of Biological Chemistry},
 number = {13}
}

@article{
 title = {CSL: A notch above the rest},
 type = {article},
 year = {2005},
 pages = {2472-2477},
 volume = {37},
 id = {951f9e11-35e5-393f-985a-6df0a62be9ff},
 created = {2023-01-10T01:46:05.458Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:05.458Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {CSL (CBF1, Suppressor of Hairless, Lag-1) is a transcription factor that is responsible for activating the genes downstream of the Notch signalling pathway, a pathway that is essential for the development of the nervous system and the differentiation of the haematopoietic system among others. In the absence of Notch signalling, CSL represses transcription of Notch target genes, and following activation by Notch, CSL is converted into a transcriptional activator and activates transcription of the same genes. These two opposing functions of CSL are mediated through interactions with distinct protein complexes. The Notch signalling pathway and its crucial cofactor CSL can maintain cells in an undifferentiated state, and have therefore been associated with a growing list of cancers. In addition, CSL has been co-opted by Epstein-Barr virus to mediate viral and host gene transcription following infection. © 2005 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Pursglove, S.E. and Mackay, J.P.},
 doi = {10.1016/j.biocel.2005.06.013},
 journal = {International Journal of Biochemistry and Cell Biology},
 number = {12}
}

@article{
 title = {Molecular mechanism of AHSP-mediated stabilization of α-hemoglobin},
 type = {article},
 year = {2004},
 pages = {629-640},
 volume = {119},
 id = {ed2d19cb-f91e-3151-b6da-974c5bb50868},
 created = {2023-01-10T01:46:06.370Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:06.370Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Hemoglobin A (HbA), the oxygen delivery system in humans, comprises two α and two β subunits. Free α-hemoglobin (αHb) is unstable, and its precipitation contributes to the pathophysiology of β thalassemia. In erythrocytes, the α-hemoglobin stabilizing protein (AHSP) binds αHb and inhibits its precipitation. The crystal structure of AHSP bound to Fe(II)-αHb reveals that AHSP specifically recognizes the G and H helices of αHb through a hydrophobic interface that largely recapitulates the α1-β1 interface of hemoglobin. The AHSP-αHb interactions are extensive but suboptimal, explaining why β-hemoglobin can competitively displace AHSP to form HbA. Remarkably, the Fe(II)-heme group in AHSP bound αHb is coordinated by the distal but not the proximal histidine. Importantly, binding to AHSP facilitates the conversion of oxy-αHb to a deoxygenated, oxidized [Fe(III)], nonreactive form in which all six coordinate positions are occupied. These observations reveal the molecular mechanisms by which AHSP stabilizes free αHb.},
 bibtype = {article},
 author = {Feng, L. and Gell, D.A. and Zhou, S. and Gu, L. and Kong, Y. and Li, J. and Hu, M. and Yan, N. and Lee, C. and Rich, A.M. and MacKay, J.P. and Shi, Y.},
 doi = {10.1016/j.cell.2004.11.025},
 journal = {Cell},
 number = {5}
}

@article{
 title = {Classic zinc finger from friend of GATA mediates an interaction with the coiled-coil of transforming acidic coiled-coil 3},
 type = {article},
 year = {2004},
 pages = {39789-39797},
 volume = {279},
 id = {8ae82118-628a-3d0f-85fb-40ef4ede1b72},
 created = {2023-01-10T01:46:07.273Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:07.273Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Classic zinc finger domains (cZFs) consist of a β-hairpin followed by an α-helix. They are among the most abundant of all protein domains and are often found in tandem arrays in DNA-binding proteins, with each finger contributing an α-helix to effect sequence-specific DNA recognition. Lone cZFs, not found in tandem arrays, have been postulated to function in protein interactions. We have studied the transcriptional co-regulator Friend of GATA (FOG), which contains nine zinc fingers. We have discovered that the third cZF of FOG contacts a coiled-coil domain in the centrosomal protein transforming acidic coiled-coil 3 (TACC3). Although FOG-ZF3 exhibited low solubility, we have used a combination of mutational mapping and protein engineering to generate a derivative that was suitable for in vitro and structural analysis. We report that the α-helix of FOG-ZF3 recognizes a C-terminal portion of the TACC3 coiled-coil. Remarkably, the α-helical surface utilized by FOG-ZF3 is the same surface responsible for the well established sequence-specific DNA-binding properties of many other cZFs. Our data demonstrate the versatility of cZFs and have implications for the analysis of many as yet uncharacterized cZF proteins.},
 bibtype = {article},
 author = {Simpson, R.J.Y. and Lee, S.H.Y. and Bartle, N. and Sum, E.Y. and Visvader, J.E. and Matthews, J.M. and Mackay, J.P. and Crossley, M.},
 doi = {10.1074/jbc.M404130200},
 journal = {Journal of Biological Chemistry},
 number = {38}
}

@article{
 title = {Presence of transient helical segments in the galanin-like peptide evident from <sup>1</sup>H NMR, circular dichroism, and prediction studies},
 type = {article},
 year = {2004},
 pages = {261-271},
 volume = {146},
 id = {dcdf6b5e-2e76-3021-abce-66a56e5b7f57},
 created = {2023-01-10T01:46:08.193Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:08.193Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods. © 2004 Elsevier Inc. All rights reserved.},
 bibtype = {article},
 author = {Dastmalchi, S. and Church, W.B. and Morris, M.B. and Iismaa, T.P. and Mackay, J.P.},
 doi = {10.1016/j.jsb.2004.01.004},
 journal = {Journal of Structural Biology},
 number = {3}
}

@article{
 title = {Structural and functional analysis of the Josephin domain of the polyglutamine protein ataxin-3},
 type = {article},
 year = {2004},
 pages = {387-394},
 volume = {322},
 id = {74e20e29-48a8-34da-ac85-4beac421f531},
 created = {2023-01-10T01:46:09.118Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:09.118Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Ataxin-3 belongs to the family of polyglutamine proteins, which are associated with nine different neurodegenerative disorders. Relatively little is known about the structural and functional properties of ataxin-3, and only recently have these aspects of the protein begun to be explored. We have performed a preliminary investigation into the conserved N-terminal domain of ataxin-3, termed Josephin. We show that Josephin is a monomeric domain which folds into a globular conformation and possesses ubiquitin protease activity. In addition, we demonstrate that the presence of the polyglutamine region of the protein does not alter the structure of the protein. However, its presence destabilizes the Josephin domain. The implications of these data in the pathogenesis of polyglutamine repeat proteins are discussed. © 2004 Elsevier Inc. All rights reserved.},
 bibtype = {article},
 author = {Chow, M.K.M. and MacKay, J.P. and Whisstock, J.C. and Scanlon, M.J. and Bottomley, S.P.},
 doi = {10.1016/j.bbrc.2004.07.131},
 journal = {Biochemical and Biophysical Research Communications},
 number = {2}
}

@article{
 title = {Crystal and Solution Structures of a Superantigen from Yersinia pseudotuberculosis Reveal a Yelly-Roll Fold},
 type = {article},
 year = {2004},
 pages = {145-156},
 volume = {12},
 id = {cff79b19-ac8f-3b35-9aa3-73d032e0181d},
 created = {2023-01-10T01:46:10.033Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:10.033Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Superantigens are a class of microbial proteins with the ability to excessively activate T cells by binding to the T cell receptor. The staphylococcal and streptococcal superantigens are closely related in structure and possess an N-terminal domain that resembles an OB fold and a C-terminal domain similar to a β-grasp fold. Yersinia pseudotuberculosis produces superantigens, YPMa, YPMb, and YPMc, which have no significant amino acid similarity to other proteins. We have determined the crystal and solution structures of YPMa, which show that the protein has a jelly-roll fold. The closest structural neighbors to YPMa are viral capsid proteins and members of the tumor necrosis factor superfamily. In the crystal structure, YPMa packs as a trimer, another feature shared with viral capsid proteins and TNF superfamily proteins. However, in solution YPMa behaves as a monomer, and any functional relevance of the trimer observed in the crystals is yet to be established.},
 bibtype = {article},
 author = {Donadini, R. and Liew, C.W. and Kwan, A.H.Y. and Mackay, J.P. and Fields, B.A.},
 doi = {10.1016/j.str.2003.12.002},
 journal = {Structure},
 number = {1}
}

@article{
 title = {Loss of α-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates β-thalassemia},
 type = {article},
 year = {2004},
 pages = {1457-1466},
 volume = {114},
 id = {b699fcb7-add4-3ff6-a946-b7bf6448eaeb},
 created = {2023-01-10T01:46:10.944Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:10.944Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Hemoglobin (Hb) A production during red blood cell development is coordinated to minimize the deleterious effects of free α- and β-Hb subunits, which are unstable and cytotoxic. The α-Hb-stabilizing protein (AHSP) is an erythroid protein that specifically binds α-Hb and prevents its precipitation in vitro, which suggests that it may function to limit free α-Hb toxicities in vivo. We investigated this possibility through gene ablation and biochemical studies. AHSP-/- erythrocytes contained hemoglobin precipitates and were short-lived. In hematopoietic tissues, erythroid precursors were elevated in number but exhibited increased apoptosis. Consistent with unstable α-Hb, AHSP-/- erythrocytes contained increased ROS and evidence of oxidative damage. Moreover, purified recombinant AHSP inhibited ROS production by α-Hb in solution. Finally, loss of AHSP worsened the phenotype of β-thalassemia, a common inherited anemia characterized by excess free α-Hb. Together, the data support a model in which AHSP binds α-Hb transiently to stabilize its conformation and render it biochemically inert prior to Hb A assembly. This function is essential for normal erythropoiesis and, to a greater extent, in β-thalassemia. Our findings raise the possibility that altered AHSP expression levels could modulate the severity of β-thalassemia in humans.},
 bibtype = {article},
 author = {Kong, Y. and Zhou, S. and Kihm, A.J. and Katein, A.M. and Yu, X. and Gell, D.A. and Mackay, J.P. and Adachi, K. and Foster-Brown, L. and Louden, C.S. and Gow, A.J. and Weiss, M.J.},
 doi = {10.1172/JCI21982},
 journal = {Journal of Clinical Investigation},
 number = {10}
}

@article{
 title = {Structural studies on a protein-binding zinc-finger domain of eos reveal both similarities and differences to classical zinc fingers},
 type = {article},
 year = {2004},
 pages = {13318-13327},
 volume = {43},
 id = {08b79ba5-e49f-31ea-9efb-6f22acd29fae},
 created = {2023-01-10T01:46:11.867Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:11.867Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The oligomerization domain that is present at the C terminus of Ikaros-family proteins and the protein Trps-1 is important for the proper regulation of developmental processes such as hematopoiesis. Remarkably, this domain is predicted to contain two classical zinc fingers (ZnFs), domains normally associated with the recognition of nucleic acids. The preference for protein binding by these predicted ZnFs is not well-understood. We have used a range of methods to gain insight into the structure of this domain. Circular dichroism, UV - vis, and NMR experiments carried out on the C-terminal domain of Eos (EosC) revealed that the two putative ZnFs (C1 and C2) are separable, i.e., capable of folding independently in the presence of ZnII. We next determined the structure of EosC2 using NMR spectroscopy, revealing that, although the overall fold of EosC2 is similar to other classical ZnFs, a number of differences exist. For example, the conformation of the C terminus of EosC2 appears to be flexible and may result in a major rearrangement of the zinc ligands. Finally, alanine-scanning mutagenesis was used to identify the residues that are involved in the homo- and hetero-oligomerization of Eos, and these results are discussed in the context of the structure of EosC. These studies provide the first structural insights into how EosC mediates protein - protein interactions and contributes to our understanding of why it does not exhibit high-affinity DNA binding.},
 bibtype = {article},
 author = {Westman, B.J. and Perdomo, J. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},
 doi = {10.1021/bi049506a},
 journal = {Biochemistry},
 number = {42}
}

@article{
 title = {TC-1 Is a Novel Tumorigenic and Natively Disordered Protein Associated with Thyroid Cancer},
 type = {article},
 year = {2004},
 pages = {2766-2773},
 volume = {64},
 id = {17d8f871-48a9-351a-8a2f-85f939c4f286},
 created = {2023-01-10T01:46:12.786Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:12.786Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {A novel gene, thyroid cancer 1 (TC-1), was found recently to be overexpressed in thyroid cancer. TC-1 shows no homology to any of the known thyroid cancer-associated genes. We have produced stable transformants of normal thyroid cells that express the TC-1 gene, and these cells show increased proliferation rates and anchorage-independent growth in soft agar. Apoptosis rates also are decreased in the transformed cells. We also have expressed recombinant TC-1 protein and have undertaken a structural and functional characterization of the protein. The protein is monomeric and predominantly unstructured under conditions of physiologic salt and pH. This places it in the category of natively disordered proteins, a rapidly expanding group of proteins, many members of which play critical roles in cell regulation processes. We show that the protein can be phosphorylated by cyclic AMP-dependent protein kinase and protein kinase C, and the activity of both of these kinases is up-regulated when cells are stably transfected with TC-1. These results suggest that overexpression of TC-1 may be important in thyroid carcinogenesis.},
 bibtype = {article},
 author = {Sunde, M. and McGrath, K.C.Y. and Young, L. and Matthews, J.M. and Chua, E.L. and Mackay, J.P. and Death, A.K.},
 doi = {10.1158/0008-5472.CAN-03-2093},
 journal = {Cancer Research},
 number = {8}
}

@article{
 title = {The C-terminal Domain of Eos Forms a High Order Complex in Solution},
 type = {article},
 year = {2003},
 pages = {42419-42426},
 volume = {278},
 id = {d44e19e1-2ac4-30ab-88a3-a1966922ed83},
 created = {2023-01-10T01:46:13.722Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:13.722Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Ikaros family transcription factors play important roles in the control of hematopoiesis. Family members are predicted to contain up to six classic zinc fingers that are arranged into N- and C-terminal domains. The N-terminal domain is responsible for site-specific DNA binding, whereas the C-terminal domain primarily mediates the homo- and hetero-oligomerization between family members. Although the mechanisms of action of these proteins are not completely understood, the zinc finger domains are known to play a central role. In the current study, we have sought to understand the physical and functional properties of these domains, in particular the C-terminal domain. We show that the N-terminal domain from Eos, and not its C-terminal region, is required to recognize GGGA consensus sequences. Surprisingly, in contrast to the behavior exhibited by Ikaros, the C-terminal domain of Eos inhibits the DNA-binding activity of the full-length protein. In addition, we have used a range of biophysical techniques to demonstrate that the C-terminal domain of Eos mediates the formation of complexes that consist of nine or ten molecules. These results constitute the first direct demonstration that Ikaros family proteins can form higher order complexes in solution, and we discuss this unexpected result in the context of what is currently known about the family members and their possible mechanism of action.},
 bibtype = {article},
 author = {Westman, B.J. and Perdomo, J. and Sunde, M. and Crossley, M. and Mackay, J.P.},
 doi = {10.1074/jbc.M306817200},
 journal = {Journal of Biological Chemistry},
 number = {43}
}

@article{
 title = {Pentaprobe: a comprehensive sequence for the one-step detection of DNA-binding activities.},
 type = {article},
 year = {2003},
 volume = {31},
 id = {82c033ec-4b64-301f-b28b-440885f5251a},
 created = {2023-01-10T01:46:14.623Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:14.623Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The rapid increase in the number of novel proteins identified in genome projects necessitates simple and rapid methods for assigning function. We describe a strategy for determining whether novel proteins possess typical sequence-specific DNA-binding activity. Many proteins bind recognition sequences of 5 bp or less. Given that there are 4(5) possible 5 bp sites, one might expect the length of sequence required to cover all possibilities would be 4(5) x 5 or 5120 nt. But by allowing overlaps, utilising both strands and using a computer algorithm to generate the minimum sequence, we find the length required is only 516 base pairs. We generated this sequence as six overlapping double-stranded oligonucleotides, termed pentaprobe, and used it in gel retardation experiments to assess DNA binding by both known and putative DNA-binding proteins from several protein families. We have confirmed binding by the zinc finger proteins BKLF, Eos and Pegasus, the Ets domain protein PU.1 and the treble clef N- and C-terminal fingers of GATA-1. We also showed that the N-terminal zinc finger domain of FOG-1 does not behave as a typical DNA-binding domain. Our results suggest that pentaprobe, and related sequences such as hexaprobe, represent useful tools for probing protein function.},
 bibtype = {article},
 author = {Kwan, A.H. and Czolij, R. and Mackay, J.P. and Crossley, M.},
 doi = {10.1093/nar/gng124},
 journal = {Nucleic acids research},
 number = {20}
}

@article{
 title = {The structure of the zinc finger domain from human splicing factor ZNF265 fold},
 type = {article},
 year = {2003},
 pages = {22805-22811},
 volume = {278},
 id = {762d5635-24ff-39d3-85a7-d0f832df931b},
 created = {2023-01-10T01:46:15.539Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:15.539Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Identification of the protein domains that are responsible for RNA recognition has lagged behind the characterization of protein-DNA interactions. However, it is now becoming clear that a range of structural motifs bind to RNA and their structures and molecular mechanisms of action are beginning to be elucidated. In this report, we have expressed and purified one of the two putative RNA-binding domains from ZNF265, a protein that has been shown to bind to the spliceosomal components U1-70K and U2AF35 and to direct alternative splicing. We show that this domain, which contains four highly conserved cysteine residues, forms a stable, monomeric structure upon the addition of 1 molar eq of Zn(II). Determination of the solution structure of this domain reveals a conformation comprising two stacked β-hairpins oriented at ·80° to each other and sandwiching the zinc ion; the fold resembles the zinc ribbon class of zinc-binding domains, although with one less β-strand than most members of the class. Analysis of the structure reveals a striking resemblance to known RNA-binding motifs in terms of the distribution of key surface residues responsible for making RNA contacts, despite a complete lack of structural homology. Furthermore, we have used an RNA gel shift assay to demonstrate that a single crossed finger domain from ZNF265 is capable of binding to an RNA message. Taken together, these results define a new RNA-binding motif and should provide insight into the functions of the >100 uncharacterized proteins in the sequence data bases that contain this domain.},
 bibtype = {article},
 author = {Plambeck, C.A. and Kwan, A.H.Y. and Adams, D.J. and Westman, B.J. and Van der Weyden, L. and Medcalf, R.L. and Morris, B.J. and Mackay, J.P.},
 doi = {10.1074/jbc.M301896200},
 journal = {Journal of Biological Chemistry},
 number = {25}
}

@article{
 title = {Structural basis for the recognition of Idb1 by the N-terminal LIM domains of LMO2 and LMO4},
 type = {article},
 year = {2003},
 pages = {2224-2233},
 volume = {22},
 id = {443069ef-b4a5-37f0-958b-48eb15aacddd},
 created = {2023-01-10T01:46:16.462Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:16.462Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {LMO2 and LMO4 are members of a small family of nuclear transcriptional regulators that are important for both normal development and disease processes. LMO2 is essential for hemopoiesis and angiogenesis, and inappropriate overexpression of this protein leads to T-cell leukemias. LMO4 is developmentally regulated in the mammary gland and has been implicated in breast oncogenesis. Both proteins comprise two tandemly repeated LIM domains. LMO2 and LMO4 interact with the ubiquitous nuclear adaptor protein Idb1/NLI/CLIM2, which associates with the LIM domains of LMO and LIM homeodomain proteins via its LIM interaction domain (Idb1-LID). We report the solution structures of two LMO:Idb1 complexes (PDB: 1M3V and 1J20) and show that Idb1-LID binds to the N-terminal LIM domain (LIM1) of LMO2 and LMO4 in an extended conformation, contributing a third strand to a β-hairpin in LIM1 domains. These findings constitute the first molecular definition of LIM-mediated protein-protein interactions and suggest a mechanism by which Idb1 can bind a variety of LIM domains that share low sequence homology.},
 bibtype = {article},
 author = {Deane, J.E. and Mackay, J.P. and Kwan, A.H.Y. and Sum, E.Y.M. and Visvader, J.E. and Matthews, J.M.},
 doi = {10.1093/emboj/cdg196},
 journal = {EMBO Journal},
 number = {9}
}

@article{
 title = {Engineering a protein scaffold from a PHD finger},
 type = {article},
 year = {2003},
 pages = {803-813},
 volume = {11},
 id = {b8887e04-25ff-3420-9eae-4d046cdc7ab2},
 created = {2023-01-10T01:46:17.361Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:17.361Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The design of proteins with tailored functions remains a relatively elusive goal. Small size, a well-defined structure, and the ability to maintain structural integrity despite multiple mutations are all desirable properties for such designer proteins. Many zinc binding domains fit this description. We determined the structure of a PHD finger from the transcriptional cofactor Mi2β and investigated the suitability of this domain as a scaffold for presenting selected binding functions. The two flexible loops in the structure were mutated extensively by either substitution or expansion, without affecting the overall fold of the domain. A binding site for the corepressor CtBP2 was also grafted onto the domain, creating a new PHD domain that can specifically bind CtBP2 both in vitro and in the context of a eukaryotic cell nucleus. These results represent a step toward designing new regulatory proteins for modulating aberrant gene expression in vivo.},
 bibtype = {article},
 author = {Kwan, A.H.Y. and Gell, D.A. and Verger, A. and Crossley, M. and Matthews, J.M. and Mackay, J.P.},
 doi = {10.1016/S0969-2126(03)00122-9},
 journal = {Structure},
 number = {7}
}

@article{
 title = {Cardiac hypertrophy and histone deacetylase-dependent transcriptional repression mediated by the atypical homeodomain protein Hop},
 type = {article},
 year = {2003},
 pages = {863-871},
 volume = {112},
 id = {ea3de67e-e52c-3b2d-8ffd-ffe82aeeec88},
 created = {2023-01-10T01:46:18.276Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:18.276Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Activation of multiple pathways is associated with cardiac hypertrophy and heart failure. Repression of antihypertrophic pathways has rarely been demonstrated to cause cardiac hypertrophy in vivo. Hop is an unusual homeodomain protein that is expressed by embryonic and postnatal cardiac myocytes. Unlike other homeodomain proteins, Hop does not bind DNA. Rather, it modulates cardiac growth and proliferation by inhibiting the transcriptional activity of serum response factor (SRF) in cardiomyocytes. Here we show that Hop can inhibit SRF-dependent transcriptional activation by recruiting histone deacetylase (HDAC) activity and can form a complex that includes HDAC2. Transgenic mice that overexpress Hop develop severe cardiac hypertrophy, cardiac fibrosis, and premature death. A mutant form of Hop, which does not recruit HDAC activity, does not induce hypertrophy. Treatment of Hop transgenic mice with trichostatin A, an HDAC inhibitor, prevents hypertrophy. In addition trichostatin A also attenuates hypertrophy induced by infusion of isoproterenol. Thus, chromatin remodeling and repression of otherwise active transcriptional processes can result in hypertrophy and heart failure, and this process can be blocked with chemical HDAC inhibitors.},
 bibtype = {article},
 author = {Kook, H. and Lepore, J.J. and Gitler, A.D. and Lu, M.M. and Yung, W.W.-M. and Mackay, J. and Zhou, R. and Ferrari, V. and Gruber, P. and Epstein, J.A.},
 doi = {10.1172/JCI19137},
 journal = {Journal of Clinical Investigation},
 number = {6}
}

@article{
 title = {CCHX zinc finger derivatives retain the ability to bind Zn(II) and mediate protein-DNA interactions},
 type = {article},
 year = {2003},
 pages = {28011-28018},
 volume = {278},
 id = {254bb9d7-8c93-373a-9c53-0d7edd74d55b},
 created = {2023-01-10T01:46:19.216Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:19.216Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Classical (CCHH) zinc fingers are among the most common protein domains found in eukaryotes. They function as molecular recognition elements that mediate specific contact with DNA, RNA, or other proteins and are composed of a ββα fold surrounding a single zinc ion that is ligated by two cysteine and two histidine residues. In a number of variant zinc fingers, the final histidine is not conserved, and in other unrelated zinc binding domains, residues such as aspartate can function as zinc ligands. To test whether the final histidine is required for normal folding and the DNA-binding function of classical zinc fingers, we focused on finger 3 of basic Krüppel-like factor. The structure of this domain was determined using NMR spectroscopy and found to constitute a typical classical zinc finger. We generated a panel of substitution mutants at the final histidine in this finger and found that several of the mutants retained some ability to fold in the presence of zinc. Consistent with this result, we showed that mutation of the final histidine had only a modest effect on DNA binding in the context of the full three-finger DNA-binding domain of basic Krüppel-like factor. Further, the zinc binding ability of one of the point mutants was tested and found to be indistinguishable from the wild-type domain. These results suggest that the final zinc chelating histidine is not an essential feature of classical zinc fingers and have implications for zinc finger evolution, regulation, and the design of experiments testing the functional roles of these domains.},
 bibtype = {article},
 author = {Simpson, R.J.Y. and Cram, E.D. and Czolij, R. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},
 doi = {10.1074/jbc.M211146200},
 journal = {Journal of Biological Chemistry},
 number = {30}
}

@article{
 title = {Solution structure and behaviour of Δ-m-α-[Ru(R,R- picchxnMe<inf>2</inf>)(phi)]<sup>2+</sup> by NMR spectroscopy and molecular modelling},
 type = {article},
 year = {2003},
 pages = {165-170},
 id = {a5866cc0-0e9f-38d7-a4df-ab0bef4ad5e5},
 created = {2023-01-10T01:46:20.125Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:20.125Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The self-association of the DNA metalloprobe Δ-cis-α-[Ru(R,R- picchxnMe2)(phi)]2+ (α-phi) in aqueous solution has been investigated using 1H NMR spectroscopy and molecular modelling. The concentration dependence of proton chemical shifts of the complex gave initial indications of a self-associated species, while its structural isomer Δ-cis-β-[Ru(R,R-picchxnMe2)(phi)]2+ (β-phi) showed no such dependence. 2D-COSY and 2D-ROESY experiments were used for the complete assignment of the proton resonances of both isomers and allowed a qualitative determination of the self-association of the a isomer through the detection of intermolecular ROEs. NMR spectroscopy can also be effectively used to differentiate Δ- and Λ-diastereomers. In addition, we show, by pulsed field gradient longitudinal eddy-current delay (PFGLED) NMR spectroscopy, that α-phi self-associates at higher concentrations with an effective molecular weight at 25 mM three times that at 2.5 mM. This apparent oligomerisation was not observed for the β-isomer. © The Royal Society of Chemistry 2003.},
 bibtype = {article},
 author = {Proudfoot, E.M. and Mackay, J.P. and Karuso, P.},
 doi = {10.1039/b208846k},
 journal = {Dalton Transactions},
 number = {2}
}

@article{
 title = {Biophysical characterization of the α-globin binding protein α-hemoglobin stabilizing protein},
 type = {article},
 year = {2002},
 pages = {40602-40609},
 volume = {277},
 id = {7bd08e1a-428f-320d-b146-14956285c9d5},
 created = {2023-01-10T01:46:21.028Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:21.028Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {α-Hemoglobin stabilizing protein (AHSP) is a small (12 kDa) and abundant erythroid-specific protein that binds specifically to free α-(hemo)globin and prevents its precipitation. When present in excess over β-globin, its normal binding partner, α-globin can have severe cytotoxic effects that contribute to important human diseases such as β-thalassemia. Because AHSP might act as a chaperone to prevent the harmful aggregation of α-globin during normal erythroid cell development and in diseases of globin chain imbalance, it is important to characterize the biochemical properties of the AHSP·α-globin complex. Here we provide the first structural information about AHSP and its interaction with α-globin. We find that AHSP is a predominantly α-helical globular protein with a somewhat asymmetric shape. AHSP and α-globin are both monomeric in solution as determined by analytical ultracentrifugation and bind each other to form a complex with 1:1 subunit stoichiometry, as judged by gel filtration and amino acid analysis. We have used isothermal titration calorimetry to show that the interaction is of moderate affinity with an association constant of 1 × 107 M-1 and is thus likely to be biologically significant given the concentration of AHSP (∼0.1 mM) and hemoglobin (∼4 mm) in the late pro-erythroblast.},
 bibtype = {article},
 author = {Dvid, G. and Yi, K. and Eaton A, S.A. and Weiss C, M.J. and Mackay E, J.P.},
 doi = {10.1074/jbc.M206084200},
 journal = {Journal of Biological Chemistry},
 number = {43}
}

@article{
 title = {A new zinc binding fold underlines the versatility of zinc binding modules in protein evolution},
 type = {article},
 year = {2002},
 pages = {639-648},
 volume = {10},
 id = {3d10c7ef-8426-3256-9aa7-a918db603a09},
 created = {2023-01-10T01:46:21.952Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:21.952Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Many different zinc binding modules have been identified. Their abundance and variety suggests that the formation of zinc binding folds might be relatively common. We have determined the structure of CH11, a 27-residue peptide derived from the first cysteine/histidine-rich region (CH1) of CREB binding protein (CBP). This peptide forms a highly ordered zinc-dependent fold that is distinct from known folds. The structure differs from a subsequently determined structure of a larger region from the CH3 region of CBP, and the CH11 fold probably represents a nonphysiologically active form. Despite this, the fold is thermostable and tolerant to both multiple alanine mutations and changes in the zinc-ligand spacing. Our data support the idea that zinc binding domains may arise frequently. Additionally, such structures may prove useful as scaffolds for protein design, given their stability and robustness.},
 bibtype = {article},
 author = {Sharpe, B.K. and Matthews, J.M. and Kwan, A.H.Y. and Newton, A. and Gell, D.A. and Crossley, M. and Mackay, J.P.},
 doi = {10.1016/S0969-2126(02)00757-8},
 journal = {Structure},
 number = {5}
}

@article{
 title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of FLIN4, an intramolecular LMO4:ldb1 complex [8]},
 type = {article},
 year = {2002},
 pages = {165-166},
 volume = {23},
 id = {d2aa00f0-07ce-3979-9e30-49c883eec666},
 created = {2023-01-10T01:46:22.869Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:22.869Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Deane, J.E. and Visvader, J.E. and Mackay, J.P. and Matthews, J.M.},
 doi = {10.1023/A:1016363414644},
 journal = {Journal of Biomolecular NMR},
 number = {2}
}

@article{
 title = {Solution structure of a hydrophobic analogue of the winter flounder antifreeze protein},
 type = {article},
 year = {2002},
 pages = {1259-1266},
 volume = {269},
 id = {e7419261-1955-324a-9050-96596e299897},
 created = {2023-01-10T01:46:23.798Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:23.798Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The solution structure of a synthetic mutant type I antifreeze protein (AFP I) was determined in aqueous solution at pH 7.0 using nuclear magnetic resonance (NMR) spectroscopy. The mutations comprised the replacement of the four Thr residues by Val and the introduction of two additional Lys-Glu salt bridges. The antifreeze activity of this mutant peptide, VVVV2KE, has been previously shown to be similar to that of the wild type protein, HPLC6 (defined here as TTTT). The solution structure reveals an ahelix bent in the same direction as the more bent conformer of the published crystalstructure of TTTT, while the side chain χ1 rotamers of VVVV2KE are similar to those of the straighter conformer in the crystal of TTTT. The Val side chains of VVVV2KE assume the same orientations as the Thr side chains of TTTT, confirming the conservative nature of this mutation. The combined data suggest that AFP I undergoes an equilibrium between straight and bent helices in solution, combined with independent equilibria between different side chain rotamers for some of the amino acid residues. The present study presents the first complete sequence-specific resonance assignments and the first complete solution structure determination by NMR of any AFP I protein.},
 bibtype = {article},
 author = {Liepinsh, E. and Otting, G. and Harding, M.M. and Ward, L.G. and Mackay, J.P. and Haymet, A.D.J.},
 doi = {10.1046/j.1432-1033.2002.02766.x},
 journal = {European Journal of Biochemistry},
 number = {4}
}

@article{
 title = {Ikaros: A key regulator of haematopoiesis},
 type = {article},
 year = {2002},
 pages = {1304-1307},
 volume = {34},
 id = {b11bff38-f90a-3ab1-9e2f-59aa100f8266},
 created = {2023-01-10T01:46:24.702Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:24.702Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function - unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription. © 2002 Elsevier Science Ltd. All rights reserved.},
 bibtype = {article},
 author = {Westman, B.J. and Mackay, J.P. and Gell, D.},
 doi = {10.1016/S1357-2725(02)00070-5},
 journal = {International Journal of Biochemistry and Cell Biology},
 number = {10}
}

@article{
 title = {Characterization of the conserved interaction between GATA and FOG family proteins},
 type = {article},
 year = {2002},
 pages = {35720-35729},
 volume = {277},
 id = {d4514158-aef2-3645-95c3-a537e76c54ce},
 created = {2023-01-10T01:46:25.606Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:25.606Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.},
 bibtype = {article},
 author = {Kowalski, K. and Liew, C.K. and Matthews, J.M. and Gell, D.A. and Crossley, M. and Mackay, J.P.},
 doi = {10.1074/jbc.M204663200},
 journal = {Journal of Biological Chemistry},
 number = {38}
}

@article{
 title = {Characterization of the conserved interaction between GATA and FOG family proteins},
 type = {article},
 year = {2002},
 pages = {35720-35729},
 volume = {277},
 id = {943dca0a-527a-3b78-8975-163e0bc0a3c9},
 created = {2023-01-10T01:46:26.825Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:26.825Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.},
 bibtype = {article},
 author = {Kowalski, K. and Liew, C.K. and Matthews, J.M. and Gell, D.A. and Crossley, M. and Mackay, J.P.},
 doi = {10.1074/jbc.M204663200},
 journal = {Journal of Biological Chemistry},
 number = {38}
}

@article{
 title = {The Bacillus subtilis cell division proteins FtsL and DivIC are intrinsically unstable and do not interact with one another in the absence of other septasomal components},
 type = {article},
 year = {2002},
 pages = {663-674},
 volume = {44},
 id = {8a1d4881-9023-34c3-97a4-9fab972fd4c2},
 created = {2023-01-10T01:46:27.729Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:27.729Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The bacterial septum appears to comprise a macromolecular assembly of essential cell division proteins (the 'septasome') that are responsible for physically dividing the cell during cytokinesis. FtsL and DivIC are essential components of this division machinery in Bacillus subtilis. We used yeast two-hybrid analysis as well as a variety of biochemical and biophysical methods to examine the proposed interaction between Bacillus subtilis FtsL and DivIC. We show that FtsL and DivIC are thermodynamically unstable proteins that are likely to be unfolded and therefore targeted for degradation unless stabilized by interactions with other components of the septasome. However, we show that this stabilization does not result from a direct interaction between FtsL and DivIC. We propose that the observed interdependence of DivIC and FtsL stability is a result of indirect interactions that are mediated by other septasomal proteins.},
 bibtype = {article},
 author = {Robson, S.A. and Michie, K.A. and Mackay, J.P. and Harry, E. and King, G.F.},
 doi = {10.1046/j.1365-2958.2002.02920.x},
 journal = {Molecular Microbiology},
 number = {3}
}

@article{
 title = {Hop is an unusual homeobox gene that modulates cardiac development},
 type = {article},
 year = {2002},
 pages = {713-723},
 volume = {110},
 id = {31c23e13-969a-3713-9fa5-129a498c180a},
 created = {2023-01-10T01:46:28.648Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:28.648Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Hop is a small, divergent homeodomain protein that lacks certain conserved residues required for DNA binding. Hop gene expression initiates early in cardiogenesis and continues in cardiomyocytes throughout embryonic and postnatal development. Genetic and biochemical data indicate that Hop functions directly downstream of Nkx2-5. Inactivation of Hop in mice by homologous recombination results in a partially penetrant embryonic lethal phenotype with severe developmental cardiac defects involving the myocardium. Inhibition of Hop activity in zebrafish embryos likewise disrupts cardiac development and results in severely impaired cardiac function. Hop physically interacts with serum response factor (SRF) and inhibits activation of SRF-dependent transcription by inhibiting SRF binding to DNA. Hop encodes an unusual homeodomain protein that modulates SRF-dependent cardiac-specific gene expression and cardiac development.},
 bibtype = {article},
 author = {Chen, F. and Kook, H. and Milewski, R. and Gitler, A.D. and Lu, M.M. and Li, J. and Nazarian, R. and Schnepp, R. and Jen, K. and Biben, C. and Mullins, M.C. and Epstein, J.A.},
 doi = {10.1016/S0092-8674(02)00932-7},
 journal = {Cell},
 number = {6}
}

@article{
 title = {Siah ubiquitin ligase is structurally related to traf and modulates tnf-α signaling},
 type = {article},
 year = {2002},
 pages = {68-75},
 volume = {9},
 id = {4c060996-79de-3f5c-ae4a-e1f05aebf808},
 created = {2023-01-10T01:46:29.556Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:29.556Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Members of the Siah (seven in absentia homolog) family of RING domain proteins are components of E3 ubiquitin ligase complexes that catalyze ubiquitination of proteins. We have determined the crystal structure of the substrate-binding domain (SBD) of murine Siah1a to 2.6 Å resolution. The structure reveals that Siah is a dimeric protein and that the SBD adopts an eight-stranded β-sandwich fold that is highly similar to the TRAF-C region of TRAF (TNF-receptor associated factor) proteins. The TRAF-C region interacts with TNF-α receptors and TNF-receptor associated death-domain (TRADD) proteins; however, our findings indicate that these interactions are unlikely to be mimicked by Siah. The Siah structure also reveals two novel zinc fingers in a region with sequence similarity to TRAF. We find that the Siah1a SBD potentiates TNF-α-mediated NF-κB activation. Therefore, Siah proteins share important similarities with the TRAF family of proteins, including their overall domain architecture, three-dimensional structure and functional activity. © 2002 Nature Publishing Group.},
 bibtype = {article},
 author = {Polekhina, G. and House, C.M. and Traficante, N. and Mackay, J.P. and Relaix, F. and Sassoon, D.A. and Parker, M.W. and Bowtell, D.D.L.},
 doi = {10.1038/nsb743},
 journal = {Nature Structural Biology},
 number = {1}
}

@article{
 title = {Type I shorthorn sculpin antifreeze protein: Recombinant synthesis, solution conformation, and ice growth inhibition studies},
 type = {article},
 year = {2002},
 pages = {24073-24080},
 volume = {277},
 id = {3b95bafc-8671-34e7-80d0-daa877523c66},
 created = {2023-01-10T01:46:30.477Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:30.477Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {A number of structurally diverse classes of "antifreeze" proteins that allow fish to survive in sub-zero ice-laden waters have been isolated from the blood plasma of cold water teleosts. However, despite receiving a great deal of attention, the one or more mechanisms through which these proteins act are not fully understood. In this report we have synthesized a type I antifreeze polypeptide (AFP) from the shorthorn sculpin Myoxocephalus scorpius using recombinant methods. Construction of a synthetic gene with optimized codon usage and expression as a glutathione S-transferase fusion protein followed by purification yielded milligram amounts of polypeptide with two extra residues appended to the N terminus. Circular dichroism and NMR experiments, including residual dipolar coupling measurements on a 15N-labeled recombinant polypeptide, show that the polypeptides are α-helical with the first four residues being more flexible than the remainder of the sequence. Both the recombinant and synthetic polypeptides modify ice growth, forming facetted crystals just below the freezing point, but display negligible thermal hysteresis. Acetylation of Lys-10, Lys-20, and Lys-21 as well as the N terminus of the recombinant polypeptide gave a derivative that displays both thermal hysteresis (0.4°C at 15 mg/ml) and ice crystal faceting. These results confirm that the N terminus of wild-type polypeptide is functionally important and support our previously proposed mechanism for all type I proteins, in which the hydrophobic face is oriented toward the ice at the ice/water interface.},
 bibtype = {article},
 author = {Fairley, K. and Westman, B.J. and Pham, L.H. and Haymet, A.D.J. and Harding, M.M. and Mackay, J.P.},
 doi = {10.1074/jbc.M200307200},
 journal = {Journal of Biological Chemistry},
 number = {27}
}

@article{
 title = {The solution structure and intramolecular associations of the Tec kinase Src homology 3 domain},
 type = {article},
 year = {2002},
 pages = {755-762},
 volume = {277},
 id = {b3f30832-cb31-30ae-a155-a163a5517400},
 created = {2023-01-10T01:46:31.373Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:31.373Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Tec is the prototypic member of a family of intracellular tyrosine kinases that includes Txk, Bmx, Itk, and Btk. Tec family kinases share similarities in domain structure with Src family kinases, but one of the features that differentiates them is a proline-rich region (PRR) preceding their Src homology (SH) 3 domain. Evidence that the PRR of Itk can bind in an intramolecular fashion to its SH3 domain and the lack of a regulatory tyrosine in the C terminus indicates that Tec kinases must be regulated by a different set of intramolecular interactions to the Src kinases. We have determined the solution structure of the Tec SH3 domain and have investigated interactions with its PRR, which contains two SH3-binding sites. We demonstrate that in vitro, the Tec PRR can bind in an intramolecular fashion to the SH3. However, the affinity is lower than that for dimerization via reciprocal PRR-SH3 association. Using site-directed mutagenesis we show that both sites can bind the Tec SH3 domain; site 1 (155KTLPPAP161) binds intramolecularly, while site 2 (165KRRPPPPIPP174) cannot and binds in an intermolecular fashion. These distinct roles for the SH3 binding sites in Tec family kinases could be important for protein targeting and enzyme activation.},
 bibtype = {article},
 author = {Pursglove, S.E. and Mulhern, T.D. and Mackay, J.P. and Hinds, M.G. and Booker, G.W.},
 doi = {10.1074/jbc.M108318200},
 journal = {Journal of Biological Chemistry},
 number = {1}
}

@article{
 title = {The hydrophobin EAS is largely unstructured in solution and functions by forming amyloid-like structures},
 type = {article},
 year = {2001},
 pages = {83-91},
 volume = {9},
 id = {f7eae917-5560-305a-8f43-505a2f44c290},
 created = {2023-01-10T01:46:32.312Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:32.312Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.},
 bibtype = {article},
 author = {Mackay, J.P. and Matthews, J.M. and Winefield, R.D. and Mackay, L.G. and Haverkamp, R.G. and Templeton, M.D.},
 doi = {10.1016/S0969-2126(00)00559-1},
 journal = {Structure},
 number = {2}
}

@article{
 title = {Discovery and Structure of a Potent and Highly Specific Blocker of Insect Calcium Channels},
 type = {article},
 year = {2001},
 pages = {40306-40312},
 volume = {276},
 id = {6c391a59-29b9-321f-858c-b5248f7b56df},
 created = {2023-01-10T01:46:33.265Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:33.265Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {We have isolated a novel family of insect-selective neurotoxins that appear to be the most potent blockers of insect voltage-gated calcium channels reported to date. These toxins display exceptional phylogenetic specificity, with at least a 10,000-fold preference for insect versus vertebrate calcium channels. The structure of one of the toxins reveals a highly structured, disulfide-rich core and a structurally disordered C-terminal extension that is essential for channel blocking activity. Weak structural/functional homology with ω-agatoxin-IVA/B, the prototypic inhibitor of vertebrate P-type calcium channels, suggests that these two toxin families might share a similar mechanism of action despite their vastly different phylogenetic specificities.},
 bibtype = {article},
 author = {Wang, X.-H. and Connor, M. and Wilson, D. and Wilson, H.I. and Nicholson, G.M. and Smith, R. and Shaw, D. and Mackay, J.P. and Alewood, P.F. and Christie, M.J. and Christie, M.J. and King, G.F.},
 doi = {10.1074/jbc.M105206200},
 journal = {Journal of Biological Chemistry},
 number = {43}
}

@article{
 title = {Design, production and characterization of FLIN2 and FLIN4: The engineering of intramolecular ldb1:LMO complexes},
 type = {article},
 year = {2001},
 pages = {493-499},
 volume = {14},
 id = {a9b6f2ec-a058-327e-abad-6aa474786e8a},
 created = {2023-01-10T01:46:34.180Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:34.180Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The nuclear LIM-only (LMO) transcription factors LMO2 and LMO4 play important roles in both normal and leukemic T-cell development. LIM domains are cysteine/histidine-rich domains that contain two structural zinc ions and that function as protein-protein adaptors; members of the LMO family each contain two closely spaced LIM domains. These LMO proteins all bind with high affinity to the nuclear protein LIM domain binding protein 1 (ldb1). The LMO-ldb1 interaction is mediated through the N-terminal LIM domain (LIM1) of LMO proteins and a 38-residue region towards the C-terminus of ldb1 [ldb1(LID)]. Unfortunately, recombinant forms of LMO2 and LMO4 have limited solubility and stability, effectively preventing structural analysis. Therefore, we have designed and constructed a fusion protein in which ldb1(LID) and LIM1 of LMO2 can form an intramolecular complex. The engineered protein, FLIN2 (fusion of the LIM interacting domain of ldb1 and the N-terminal LIM domain of LMO2) has been expressed and purified in milligram quantities. FLIN2 is monomeric, contains significant levels of secondary structure and yields a sharp and well-dispersed one-dimensional 1H NMR spectrum. The analogous LMO4 protein, FLIN4, has almost identical properties. These data suggest that we will be able to obtain high-resolution structural information about the LMO-ldb1 interactions.},
 bibtype = {article},
 author = {Deane, J.E. and Sum, E. and Mackay, J.P. and Lindeman, G.J. and Visvader, J.E. and Matthews, J.M.},
 doi = {10.1093/protein/14.7.493},
 journal = {Protein Engineering},
 number = {7}
}

@article{
 title = {<sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of FLIN2, an intramolecular LMO2:ldb1 complex [2]},
 type = {article},
 year = {2001},
 pages = {385-386},
 volume = {21},
 id = {4480e900-209e-3853-9615-5c2f38cb7107},
 created = {2023-01-10T01:46:35.100Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:35.100Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Matthews, J.M. and Visvader, J.E. and Mackay, J.P.},
 doi = {10.1023/A:1013373203772},
 journal = {Journal of Biomolecular NMR},
 number = {4}
}

@article{
 title = {Probing site specificity of DNA binding metallointercalators by NMR spectroscopy and molecular modeling},
 type = {article},
 year = {2001},
 pages = {4867-4878},
 volume = {40},
 id = {ecd0710f-2f62-38e1-9576-54331818c01f},
 created = {2023-01-10T01:46:35.999Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:35.999Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The molecular recognition of oligonucleotides by chiral ruthenium complexes has been probed by NMR spectroscopy using the template Δ-cis-α- and Δ-cis-β-[Ru(RR-picchxnMe2) (bidentate)]2+, where the bidentate ligand is one of phen (1,10-phenanthroline), dpq (dipyrido[3,2-f:2′,3′-h]quinoxaline), or phi (9,10-phenanthrenequinone diimine) and picchxnMe2 is N,N′-dimethyl-N,N′-di(2-picolyl)-1,2-diaminocyclohexane. By varying only the bidentate ligand in a series of complexes, it was shown that the bidentate alone can alter binding modes. DNA binding studies of the Δ-cis-α,-[Ru(RR-picchxnMe2)(phen)]2+ complex indicate fast exchange kinetics on the chemical shift time scale and a "partial intercalation" mode of binding. This complex binds to [d(CGCGATCGCG)]2 and [d(ATATCGATAT)]2 at AT, TA, and GA sites from the minor groove, as well as to the ends of the oligonucleotide at low temperature. Studies of the Δ-cis-β-[Ru(RR-picchxnMe2)(phen)]2+ complex with [d(CGCGATCGCG)]2 showed that the complex binds only weakly to the ends of the oligonucleotide. The interaction of Δ-cis-α-[Ru(RR-picchxnMe2)-(dpq)]2+ with [d(CGCGATCGCG)]2 showed intermediate exchange kinetics and evidence of minor groove intercalation at the GA base step. In contrast to the phen and dpq complexes, Δ-cis-α- and Δ-cis-β-[Ru-(RR-picchxnMe2)(phi)]2+ showed evidence of major groove binding independent of the metal ion configuration. DNA stabilization induced by complex binding to [d(CGCGATCGCG)]2 (measured as ΔTm) increases in the order phen < dpq and DNA affinity in the order phen < dpq < phi. The groove binding preferences exhibited by the different bidentate ligands is explained with the aid of molecular modeling experiments.},
 bibtype = {article},
 author = {Proudfoot, E.M. and Mackay, J.P. and Karuso, P.},
 doi = {10.1021/bi001655f},
 journal = {Biochemistry},
 number = {15}
}

@article{
 title = {The N-terminal Zinc Finger of the Erythroid Transcription Factor GATA-1 Binds GATC Motifs in DNA},
 type = {article},
 year = {2001},
 pages = {35794-35801},
 volume = {276},
 id = {668a6be7-8063-3f78-a869-df88d49978b0},
 created = {2023-01-10T01:46:36.902Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:36.902Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The mammalian transcription factor GATA-1 is required for normal erythroid and megakaryocytic development. GATA-1 contains two zinc fingers, the C-terminal finger, which is known to bind (A/T)GATA(A/G) motifs in DNA and the N-finger, which is important for interacting with co-regulatory proteins such as Friend of GATA (FOG). We now show that, like the C-finger, the N-finger of GATA-1 is also capable of binding DNA but recognizes distinct sequences with the core GATC. We demonstrate that the GATA-1 N-finger can bind these sequences in vitro and that in cellular assays, GATA-1 can activate promoters containing GATC motifs. Experiments with mutant GATA-1 proteins confirm the importance of the N-finger, as the C-finger is not required for transactivation from GATC sites. Recently four naturally occurring mutations in GATA-1 have been shown to be associated with familial blood disorders. These mutations all map to the N-finger domain. We have investigated the effect of these mutations on the recognition of GATC sites by the N-finger and show that one mutation R216Q abolishes DNA binding, whereas the others have only minor effects.},
 bibtype = {article},
 author = {Newton, A. and Mackay, J. and Crossley, M.},
 doi = {10.1074/jbc.M106256200},
 journal = {Journal of Biological Chemistry},
 number = {38}
}

@article{
 title = {The transactivation domain within cysteine/histidine-rich region 1 of CBP comprises two novel zinc-binding modules},
 type = {article},
 year = {2000},
 pages = {15128-15134},
 volume = {275},
 id = {912798e3-760b-3e6e-bdd0-d9324be5e544},
 created = {2023-01-10T01:46:37.833Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:37.833Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {cAMP-response element-binding protein-binding protein (CBP) is a transcriptional coactivator that interacts with a number of DNA-binding proteins and cofactor proteins involved in the regulation of transcription. Relatively little is known about the structure of CBP, but it has been noted that it contains three domains that are rich in cysteine and histidine (CH1, CH2, and CH3). The sequence of CH2 conforms to that of a leukemia-associated protein domain (PHD finger), and it has been postulated that this and both CH1 and CH3 may be zinc finger domains. This has not, however, been demonstrated experimentally. We have studied CH1 and show that it is composed of two novel zinc-binding modules, which we term 'zinc bundles.' Each bundle contains the sequence Cys-X4-Cys-X8-His-X3-Cys, and we show that a synthetic peptide comprising one zinc bundle from CH1 can fold in a zinc- dependent manner. CH3 also appears to contain two zinc bundles, one with the variant sequence Cys-X2-Cys-X9-His-X3-Cys, and we demonstrate that this variant motif also undergoes Zn(II)-induced folding. CH1 acts as a transcriptional activation domain in cellular assays. We show that mutations in any of the four zinc-chelating residues in either zinc bundle of CH1 significantly impair this activity and that these mutations also interfere with certain protein-protein interactions mediated by CH1. Our results indicate that CBP is a genuine zinc-binding protein and introduce zinc bundles as novel protein interaction domains.},
 bibtype = {article},
 author = {Newton, A.L. and Sharped, B.K. and Kwan, A. and Mackay, J.P. and Crossley, M.},
 doi = {10.1074/jbc.M910396199},
 journal = {Journal of Biological Chemistry},
 number = {20}
}

@article{
 title = {The core of the respiratory syncytial virus fusion protein is a trimeric coiled coil},
 type = {article},
 year = {2000},
 pages = {5911-5920},
 volume = {74},
 id = {82505942-a661-3722-bf77-e61b0007b83b},
 created = {2023-01-10T01:46:38.745Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:38.745Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Entry into the host cell by enveloped viruses is mediated by fusion (F) or transmembrane glycoproteins. Many of these proteins share a fold comprising a trimer of antiparallel coiled-coil heterodimers, where the heterodimers are formed by two discontinuous heptad repeat motifs within the proteolytically processed chain. The F protein of human respiratory syncytial virus (RSV; the major cause of lower respiratory tract infections in infants) contains two corresponding regions that are predicted to form coiled coils (HR1 and HR2), together with a third predicted heptad repeat (HR3) located in a nonhomologous position. In order to probe the structures of these three domains and ascertain the nature of the interactions between them, we have studied the isolated HR1, HR2, and HR3 domains of RSV F by using a range of biophysical techniques, including circular dichroism, nuclear magnetic resonance spectroscopy, and sedimentation equilibrium. HR1 forms a symmetrical, trimeric coiled coil in solution (K3 ≃ 2.2 x 1011 M-2) which interacts with HR2 to form a 3:3 hexamer. The HR1-HR2 interaction domains have been mapped using limited proteolysis, reversed-phase high- performance liquid chromatography, and electrospray-mass spectrometry. HR2 in isolation exists as a largely unstructured monomer, although it exhibits a tendency to form aggregates with β-sheet-like characteristics. Only a small increase in α-helical content was observed upon the formation of the hexamer. This suggests that the RSV F glycoprotein contains a domain that closely resembles the core structure of the simian parainfluenza virus 5 fusion protein (K. A. Baker, R. E. Dutch, R. A. Lamb, and T. S. Jardetzky, Mol. Cell 3:309-319, 1999). Finally, HR3 forms weak α-helical homodimers that do not appear to interact with HR1, HR2, or the HR1-HR2 complex. The results of these studies support the idea that viral fusion proteins have a common core architecture.},
 bibtype = {article},
 author = {Matthews, J.M. and Young, T.F. and Tucker, S.P. and Mackay, J.P.},
 doi = {10.1128/JVI.74.13.5911-5920.2000},
 journal = {Journal of Virology},
 number = {13}
}

@article{
 title = {Solution structures of two CCHC zinc fingers from the FOG family protein U-shaped that mediate protein-protein interactions},
 type = {article},
 year = {2000},
 pages = {1157-1166},
 volume = {8},
 id = {4220c92e-ca96-3078-a476-5f478fe44cdb},
 created = {2023-01-10T01:46:39.669Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:39.669Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.},
 bibtype = {article},
 author = {Chu Kong Liew, undefined and Kowalski, K. and Fox, A.H. and Newton, A. and Sharpe, B.K. and Crossley, M. and Mackay, J.P.},
 doi = {10.1016/S0969-2126(00)00527-X},
 journal = {Structure},
 number = {11}
}

@article{
 title = {Interfacial asparagine residues within an amide tetrad contribute to Max helix-loop-helix leucine zipper homodimer stability},
 type = {article},
 year = {2000},
 pages = {37454-37461},
 volume = {275},
 id = {44466ab6-2aa6-3104-b086-fd106c65ff76},
 created = {2023-01-10T01:46:40.592Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:40.592Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The transcription factor Max is the obligate dimerization partner of the Myc oncoprotein. The pivotal role of Max within the Myc regulatory network is dependent upon its ability to dimerize via the helix-loop-helix leucine zipper domain. The Max homodimer contains a tetrad of polar residues at the interface of the leucine zipper domain. A conserved interfacial Asn residue at an equivalent position in two other leucine zipper proteins has been shown to decrease homodimer stability. The unusual arrangement of this Gln-Asn/Gln'-Asn' tetrad prompted us to investigate whether Asn92 plays a similar role in destabilizing the Max homodimer. This residue was sequentially replaced with aliphatic and charged residues. Thermal denaturation, redox time course and analytical ultracentrifugation studies show that the N92V mutation does not increase homodimer stability. Replacing this residue with negatively charged side chains in N92D and N92E destabilizes the mutant homodimer. Further replacement of Gln91 indicated that H bonding between Gln91 and Asn92 residues is not significant to the stability of the native protein. These data collectively demonstrate the central role of Asn92 in homodimer interactions. Molecular modelling studies illustrate the favorable packing of the native Asn residue at the dimer interface compared with that of the mutant Max peptides.},
 bibtype = {article},
 author = {Tchan, M.C. and Choy, K.J. and Mackay, J.P. and Lyons, A.T.L. and Bains, N.P.S. and Weiss, A.S.},
 doi = {10.1074/jbc.M004264200},
 journal = {Journal of Biological Chemistry},
 number = {48}
}

@article{
 title = {A class of zinc fingers involved in protein-protein interactions},
 type = {article},
 year = {2000},
 pages = {1030-1038},
 volume = {267},
 id = {3aade15c-920e-3fd6-9d2d-2812368f8f88},
 created = {2023-01-10T01:46:41.497Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:41.497Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Zinc fingers (ZnFs) are extremely common protein domains. Several classes of ZnFs are distinguished by the nature and spacing of their zinc- coordinating residues. While the structure and function of some ZnFs are well characterized, many others have been identified only through their amino acid sequence. A number of proteins contain a conserved C-X2-C-X12-H-X1-5-C sequence, which is similar to the spacing observed for the 'classic' CCHH ZnFs. Although these domains have been implicated in protein-protein (and not protein-nucleic acid) interactions, nothing is known about their structure or function at a molecular level. Here, we address this problem through the expression and biophysical characterization of several CCHC-type zinc fingers from the erythroid transcription factor FOG and the related Drosophila protein U-shaped. Each of these domains does indeed fold in a zinc-dependent fashion, coordinating the metal in a tetrahedral manner through the sidechains of one histidine and three cysteine residues, and forming extremely thermostable structures. Analysis of CD spectra suggests an overall fold similar to that of the CCHH fingers, and indeed a point mutant of FOG-F1 in which the final cysteine residue is replaced by histidine remains capable of folding. However, the CCHC (as opposed to CCHH) motif is a prerequisite for GATA-1 binding activity, demonstrating that CCHC and CCHH topologies are not interchangeable. This demonstration that members of a structurally distinct subclass of genuine zinc finger domains are involved in the mediation of protein-protein interactions has implications for the prediction of protein function from nucleotide sequences.},
 bibtype = {article},
 author = {Matthews, J.M. and Kowalski, K. and Liew, C.K. and Sharpe, B.K. and Fox, A.H. and Crossley, M. and Mackay, J.P.},
 doi = {10.1046/j.1432-1327.2000.01095.x},
 journal = {European Journal of Biochemistry},
 number = {4}
}

@article{
 title = {The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG},
 type = {article},
 year = {1999},
 pages = {249-262},
 volume = {13},
 id = {649a742c-0f2a-3975-8e3d-1abdb42ec917},
 created = {2023-01-10T01:46:42.416Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:42.416Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted β-hairpins and a single α-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA- 1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.},
 bibtype = {article},
 author = {Kowalski, K. and Czolij, R. and King, G.F. and Crossley, M. and Mackay, J.P.},
 doi = {10.1023/A:1008309602929},
 journal = {Journal of Biomolecular NMR},
 number = {3}
}

@article{
 title = {The dimerization and topological specificity functions of MinE reside in a structurally autonomous C-terminal domain},
 type = {article},
 year = {1999},
 pages = {1161-1169},
 volume = {31},
 id = {cd93d5ad-9177-3466-a3fb-e5c4f562db3d},
 created = {2023-01-10T01:46:43.487Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:43.487Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Correct placement of the division septum in Escherichia coli requires the co-ordinated action of three proteins, MinC, MinD and MinE. MinC and MinD interact to form a non-specific division inhibitor that blocks septation at all potential division sites. MinE is able to antagonize MinCD in a topologically sensitive manner, as it restricts MinCD activity to the unwanted division sites at the cell poles. Here, we show that the topological specificity function of MinE residues in a structurally autonomous, trypsin-resistant domain comprising residues 31-88. Nuclear magnetic resonance (NMR) and circular dichroic spectroscopy indicate that this domain includes both α and β secondary structure, while analytical ultracentrifugation reveals that it also contains a region responsible for MinE homodimerization. While trypsin digestion indicates that the anti-MinCD domain of MinE (residues 1-22) does not form a tightly folded structural domain, NMR analysis of a peptide corresponding to MinE1-22 indicates that this region forms a nascent helix in which the peptide rapidly interconverts between disordered (random coil) and α-helical conformations. This suggests that the N-terminal region of MinE may be poised to adopt an α-helical conformation when it interacts with the target of its anti-MinCD activity, presumably MinD.},
 bibtype = {article},
 author = {King, G.F. and Rowland, S.L. and Pan, B. and Mackay, J.P. and Mullen, G.P. and Rothfield, L.I.},
 doi = {10.1046/j.1365-2958.1999.01256.x},
 journal = {Molecular Microbiology},
 number = {4}
}

@article{
 title = {Autonomous folding of a peptide corresponding to the N-terminal β- hairpin from ubiquitin},
 type = {article},
 year = {1999},
 pages = {1320-1331},
 volume = {8},
 id = {2fa2cd79-9bc1-3f5c-8753-6943e5016588},
 created = {2023-01-10T01:46:44.399Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:44.399Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The N-terminal 17 residues of ubiquitin have been shown by 1H NMR to fold autonomously into a β-hairpin structure in aqueous solution. This structure has a specific, native-like register, though side-chain contacts differ in detail from those observed in the intact protein. An autonomously folding hairpin has previously been identified in the case of streptococcal protein G, which is structurally homologous with ubiquitin, but remarkably, the two are not in topologically equivalent positions in the fold. This suggests that the organization of folding may be quite different for proteins sharing similar tertiary structures. Two smaller peptides have also been studied, corresponding to the isolated arms of the N-terminal hairpin of ubiquitin, and significant differences from simple random coil predictions observed in the spectra of these subfragments, suggestive of significant limitation of the backbone conformational space sampled, presumably as a consequence of the strongly β-structure favoring composition of the sequences. This illustrates the ability of local sequence elements to express a propensity for β-structure even in the absence of actual sheet formation. Attempts were made to estimate the population of the folded state of the hairpin, in terms of a simple two-state folding model. Using published 'random coil' values to model the unfolded state, and values derived from native ubiquitin for the putative unique, folded state, it was found that the apparent population varied widely for different residues and with different NMR parameters. Use of the spectra of the subfragment peptides to provide a more realistic model of the unfolded state led to better agreement in the estimates that could be obtained from chemical shift and coupling constant measurements, while making it clear that some other approaches to population estimation could not give meaningful results, because of the tendency to populate the β-region of conformational space even in the absence of the hairpin structure.},
 bibtype = {article},
 author = {Zerella, R. and Evans, P.A. and Ionides, J.M.C. and Packman, L.C. and Trotter, B.W. and Mackay, J.P. and Williams, D.H.},
 doi = {10.1110/ps.8.6.1320},
 journal = {Protein Science},
 number = {6}
}

@article{
 title = {Transcriptional cofactors of the FOG family interact with GATA proteins by means of multiple zinc fingers},
 type = {article},
 year = {1999},
 pages = {2812-2822},
 volume = {18},
 id = {008d2a64-1e9c-3d79-90e8-1d4d6afcbc71},
 created = {2023-01-10T01:46:45.293Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:45.293Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Friend of GATA-1 (FOG-1) is a zinc finger protein that has been shown to interact physically with the erythroid DNA-binding protein GATA-1 and modulate its transcriptional activity. Recently, two new members of the FOG family have been identified: a mammalian protein, FOG-2, that also associates with GATA-1 and other mammalian GATA factors; and U-shaped, a Drosophila protein that interacts with the Drosophila GATA protein Pannier. FOG proteins contain multiple zinc fingers and it has been shown previously that the sixth finger of FOG-1 interacts specifically with the N-finger but not the C-finger of GATA-1. Here we show that fingers 1, 5 and 9 of FOG-1 also interact with the N-finger of GATA-1 and that FOG-2 and U-shaped also contain multiple GATA-interacting fingers. We define the key contact residues and show that these residues are highly conserved in GATA-interacting fingers. We examine the effect of selectively mutating the four interacting fingers of FOG-1 and show that each contributes to FOG-1's ability to modulate GATA-1 activity. Finally, we show that FOG-1 can repress GATA-1-mediated activation and present evidence that this ability involves the recently described CtBP co-repressor proteins that recognize all known FOG proteins.},
 bibtype = {article},
 author = {Fox, A.H. and Liew, C. and Holmes, M. and Kowalski, K. and Mackay, J. and Crossley, M.},
 doi = {10.1093/emboj/18.10.2812},
 journal = {EMBO Journal},
 number = {10}
}

@article{
 title = {Use of altered specificity mutants to probe a specific protein-protein interaction in differentiation: The GATA-1:FOG complex},
 type = {article},
 year = {1999},
 pages = {219-228},
 volume = {3},
 id = {ed2ce0ef-2974-32de-ac4b-b652a7d61a3b},
 created = {2023-01-10T01:46:46.200Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:46.200Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {GATA-1 and FOG (Friend of GATA-1) are each essential for erythroid and megakaryocyte development. FOG, a zinc finger protein, interacts with the amino (N) finger of GATA-1 and cooperates with GATA-1 to promote differentiation. To determine whether this interaction is critical for GATA- 1 action, we selected GATA-1 mutants in yeast that fail to interact with FOG but retain normal DNA binding, as well a compensatory FOG mutant that restores interaction. These novel GATA-1 mutants do not promote erythroid differentiation of GATA-1- erythroid cells. Differentiation is rescued by the second-site FOG mutant. Thus, interaction of FOG with GATA-1 is essential for the function of GATA-1 in erythroid differentiation. These findings provide a paradigm for dissecting protein-protein associations involved in mammalian development.},
 bibtype = {article},
 author = {Crispino, J.D. and Lodish, M.B. and MacKay, J.P. and Orkin, S.H.},
 doi = {10.1016/S1097-2765(00)80312-3},
 journal = {Molecular Cell},
 number = {2}
}

@article{
 title = {The C-terminal region of the stalk domain of ubiquitous human kinesin heavy chain contains the binding site for kinesin light chain},
 type = {article},
 year = {1998},
 pages = {16663-16670},
 volume = {37},
 id = {b73ee519-2d24-3501-8e20-4c8b4ead145e},
 created = {2023-01-10T01:46:47.102Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:47.102Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The motor protein kinesin is a heterotetramer composed of two heavy chains of ~120 kDa and two light chains of ~6 kDa protein. Kinesin motor activity is dependent on the presence of ATP and microtubules. The kinesin light chain-binding site in human kinesin heavy chain was determined by reconstituting in vitro a complex of recombinant heavy and light chains. The proteins expressed in bacteria included oligohistidine-tagged fragments of human ubiquitous kinesin heavy chain, spanning most of the stalk and all of the tail domain (amino acids 555-963); and untagged, essentially full-length human kinesin light chain (4-569) along with N-terminal (4-363)and C-terminal (364-569) light chain fragments. Heavy chain fragments were attached to Ni2+-charged beads and incubated with untagged light chain fragments. Analysis of eluted complexes by SDS-PAGE and immunoblotting mapped the light chain-binding site in heavy chain to amino acids 771-813, a region close to the C-terminal end of the heavy chain stalk domain. In addition, only the full-length and N-terminal kinesin light chain fragments bound to this heavy chain region. Within this heavy chain region are four highly conserved contiguous heptad repeats (775-802) which are predicted to form a tight α- helical coiled-coil interaction with the heptad repeat-containing N-terminus of the light chain, in particular region 106-152 of human light chain. This predicted hydrophobic, α-helical coiled-coil interaction is supported by both circular dichroism spectroscopy of the recombinant kinesin heavy chain fragment 771-963, which displays an α-helical content of 70%, and the resistance of the heavy/light chain interdiction to high salt (0.5 M).},
 bibtype = {article},
 author = {Diefenbach, R.J. and Mackay, J.P. and Armati, P.J. and Cunningham, A.L.},
 doi = {10.1021/bi981163r},
 journal = {Biochemistry},
 number = {47}
}

@article{
 title = {Key residues characteristic of GATA N-fingers are recognized by FOG},
 type = {article},
 year = {1998},
 pages = {33595-33603},
 volume = {273},
 id = {e9e15c4c-0ebf-3abb-b74a-16640859e1ae},
 created = {2023-01-10T01:46:48.015Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:48.015Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Protein-protein interactions play significant roles in the control of gene expression. These interactions often occur between small, discrete domains within different transcription factors. In particular, zinc fingers, usually regarded as DNA-binding domains, are now also known to be involved in mediating contacts between proteins. We have investigated the interaction between the erythroid transcription factor GATA-1 and its partner, the 9 zinc finger protein, FOG (Friend Of GATA). We demonstrate that this interaction represents a genuine finger-finger contact, which is dependent on zinc- coordinating residues within each protein. We map the contact domains to the core of the N-terminal zinc finger of GATA-1 and the 6th zinc finger of FOG. Using a scanning substitution strategy we identify key residues within the GATA-1 N-finger which are required for FOG binding. These residues are conserved in the N-fingers of all GATA proteins known to bind FOG, but are not found in the respective C-fingers. This observation may, therefore, account for the particular specificity of FOG for N-fingers. Interestingly, the key N-finger residues are seen to form a contiguous surface, when mapped onto the structure of the N-finger of GATA-1.},
 bibtype = {article},
 author = {Fox, A.H. and Kowalski, K. and King, G.F. and Mackay, J.P. and Crossley, M.},
 doi = {10.1074/jbc.273.50.33595},
 journal = {Journal of Biological Chemistry},
 number = {50}
}

@article{
 title = {Zinc fingers are sticking together},
 type = {article},
 year = {1998},
 pages = {1-4},
 volume = {23},
 id = {76060f13-2fda-31b9-b159-68b649adeb20},
 created = {2023-01-10T01:46:48.915Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:48.915Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 bibtype = {article},
 author = {Mackay, J.P. and Crossley, M.},
 doi = {10.1016/S0968-0004(97)01168-7},
 journal = {Trends in Biochemical Sciences},
 number = {1}
}

@article{
 title = {Interactions of the Antitumor Agent Molybdocene Dichloride with Oligonucleotides},
 type = {article},
 year = {1998},
 pages = {2432-2437},
 volume = {37},
 id = {3622d446-6dc4-3dca-890e-5982c68ebdd0},
 created = {2023-01-10T01:46:49.823Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:49.823Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The interactions between the antitumor-active metallocene molybdocene dichloride (Cp2MoCl2) and four oligonucleotides have been studied by 1H and 31P NMR spectroscopy. In 50 mM salt solutions of molybdocene dichloride, hydrolysis of the halide ligands occurs to give a solution with pD 2, containing a species in which both Cp rings remain metal bound for 24 h. At pD 7, however, partial hydrolysis of the Cp rings (∼30%) occurs after 24 h. Addition of an aqueous solution of molybdocene dichloride in 50 mM salt to the self-complementary sequence d(CGCATATGCG)2, maintaining the pD at 6.0-7.0, showed no evidence for the formation of a metallocene-oligonucleotide complex, and only peaks arising from hydrolysis of molybdocene dichloride were detected. A similar result was obtained in titration experiments with the single-stranded sequence d(ATGGTA) at pD 6.5-7.0. However, at pD 3.0, new signals assigned to a molybdocene-oligonuleotide complex or complexes were detected in the 1H NMR spectrum. No change was observed in the 31P NMR spectrum. The complex or complexes formed between molybdocene dichloride and d(ATGGTA) are stable for hours at pD 3.0; at higher pD, the complex is destabilized and only peaks arising from hydrolysis of molybdocene dichloride are detected. Titration experiments at low pD with the dinucleotide dCpG showed a new set of signals in the 1H NMR spectrum, tentatively assigned to formation of a complex arising due to coordination of molybdenum to guanine N7 and/or cytosine N3. At pD 7.0, these signals disappeared. The results obtained show that stable oligonucleotide adducts are not formed in 50 mM salt at pD 7.0, and hence it is highly unlikely that formation of molybdocene - DNA adducts in vivo is the primary action that is responsible for the antitumor properties of molybdocene dichloride.},
 bibtype = {article},
 author = {Harding, M.M. and Mokdsi, G. and Mackay, J.P. and Prodigalidad, M. and Lucas, S.W.},
 doi = {10.1021/ic971205k},
 journal = {Inorganic Chemistry},
 number = {10}
}

@article{
 title = {Involvement of the N-finger in the self-association of GATA-1},
 type = {article},
 year = {1998},
 pages = {30560-30567},
 volume = {273},
 id = {4c338f22-0421-34a2-92f2-d97f844ae7db},
 created = {2023-01-10T01:46:50.749Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:50.749Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Zinc fingers are recognized as small protein domains that bind to specific DNA sequences. Recently however, zinc fingers from a number of proteins, in particular the GATA family of transcription factors, have also been implicated in specific protein-protein interactions. The erythroid protein GATA-1 contains two zinc fingers: the C-finger, which is sufficient for sequence-specific DNA-binding, and the N-finger, which appears both to modulate DNA-binding and to interact with other transcription factors. We have expressed and purified the N-finger domain and investigated its involvement in the self-association of GATA-1. We demonstrate that this domain does not homodimerize but instead makes intermolecular contacts with the C-finger, suggesting that GATA dimers are maintained by reciprocal N- finger-C-finger contacts. Deletion analysis identifies a 25-residue region, C-terminal to the core N-finger domain, that is sufficient for interaction with intact GATA-1. A similar subdomain exists C-terminal to the C-finger, and we show that self-association is substantially reduced when both subdomains are disrupted by mutation. Moreover, mutations that impair GATA-1 self-association also interfere with its ability to activate transcription in transfection studies.},
 bibtype = {article},
 author = {Mackay, J.P. and Kowalski, K. and Fox, A.H. and Czolij, R. and King, G.F. and Crossley, M.},
 doi = {10.1074/jbc.273.46.30560},
 journal = {Journal of Biological Chemistry},
 number = {46}
}

@article{
 title = {Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides},
 type = {article},
 year = {1997},
 pages = {67-77},
 volume = {4},
 id = {e3e2a974-5c9b-3e6e-8d6b-ec39b861bc0a},
 created = {2023-01-10T01:46:51.663Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:51.663Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides. © 1997 ESCOM Science Publishers B.V.},
 bibtype = {article},
 author = {Bains, N.P.S. and Wilce, J.A. and Heuer, K.H. and Tunstall, M. and Mackay, J.P. and Bennett, M.R. and Weiss, A.S. and King, G.F.},
 doi = {10.1007/BF02443517},
 journal = {International Journal of Peptide Research and Therapeutics},
 number = {2}
}

@article{
 title = {Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR},
 type = {article},
 year = {1997},
 pages = {1-8},
 volume = {10},
 id = {53beac46-6091-3dfa-988a-801d2a1ac05f},
 created = {2023-01-10T01:46:52.607Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:52.607Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.},
 bibtype = {article},
 author = {Dingley, A.J. and Mackay, J.P. and Shaw, G.L. and Hambly, B.D. and King, G.F.},
 doi = {10.1023/A:1018339526108},
 journal = {Journal of Biomolecular NMR},
 number = {1}
}

@article{
 title = {The structure of versutoxin (δ-atracotoxin-Hv1) provides insights into the binding of site 3 neurotoxins to the voltage-gated sodium channel},
 type = {article},
 year = {1997},
 pages = {1525-1535},
 volume = {5},
 id = {c641551b-c512-3935-9b0e-32d0d876b55e},
 created = {2023-01-10T01:46:53.511Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:53.511Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background: Versutoxin (δ-ACTX-Hv1) is the major component of the venom of the Australian Blue Mountains funnel web spider, Hadronyche versuta. δ-ACTX-Hv1 produces potentially fatal neurotoxic symptoms in primates by slowing the inactivation of voltage-gated sodium channels; δ-ACTX-Hv1 is therefore a useful tool for studying sodium channel function. We have determined the three-dimensional structure of δ-ACTX-Hv1 as the first step towards understanding the molecular basis of its interaction with these channels. Results: The solution structure of δ-ACTX-Hv1, determined using NMR spectroscopy, comprises a core β region containing a triple-stranded antiparallel β sheet, a thumb-like extension protruding from the β region and a C-terminal 310 helix that is appended to the β domain by virtue of a disulphide bond. The β region contains a cystine knot motif similar to that seen in other neurotoxic polypeptides. The structure shows homology with μ-agatoxin-I, a spider toxin that also modifies the inactivation kinetics of vertebrate voltage-gated sodium channels. More surprisingly, δ-ACTX-Hv1 shows both sequence and structural homology with gurmarin, a plant polypeptide. This similarity leads us to suggest that the sweet-taste suppression elicited by gurmarin may result from an interaction with one of the downstream ion channels involved in sweet-taste transduction. Conclusions: δ-ACTX-Hv1 shows no structural homology with either sea anemone or α-scorpion toxins, both of which also modify the inactivation kinetics of voltage-gated sodium channels by interacting with channel recognition site 3. However, we have shown that δ-ACTX-Hv1 contains charged residues that are topologically related to those implicated in the binding of sea anemone and α-scorpion toxins to mammalian voltage-gated sodium channels, suggesting similarities in their mode of interaction with these channels.},
 bibtype = {article},
 author = {Fletcher, J.I. and Chapman, B.E. and Mackay, J.P. and Howden, M.E.H. and King, G.F.},
 doi = {10.1016/S0969-2126(97)00301-8},
 journal = {Structure},
 number = {11}
}

@article{
 title = {Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides},
 type = {article},
 year = {1997},
 pages = {67-77},
 volume = {4},
 id = {424daf6d-00c1-3bcf-ad60-9abdb95c2e4b},
 created = {2023-01-10T01:46:54.417Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:54.417Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides.},
 bibtype = {article},
 author = {Bains, N.P.S. and Wilce, J.A. and Heuer, K.H. and Tunstall, M. and Mackay, J.P. and Bennett, M.R. and Weiss, A.S. and King, G.F.},
 doi = {10.1007/BF02443517},
 journal = {Letters in Peptide Science},
 number = {2}
}

@article{
 title = {DMA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II},
 type = {article},
 year = {1997},
 pages = {387-398},
 volume = {42},
 id = {964a530f-b34f-3e6c-917f-3ea08bb03126},
 created = {2023-01-10T01:46:55.338Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:55.338Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley & Sons, Inc.},
 bibtype = {article},
 author = {Harding, M.M. and Krippner, G.Y. and Shelton, C.J. and Rodger, A. and Sanders, K.J. and Mackay, J.P. and Prakash, A.S.},
 doi = {10.1002/(SICI)1097-0282(19971005)42:4<387::AID-BIP2>3.0.CO;2-M},
 journal = {Biopolymers - Nucleic Acid Sciences Section},
 number = {4}
}

@article{
 title = {Δ-cis-α-[Ru(RR-picchxnMe<inf>2</inf>)(phen)]<sup>2+</sup> shows minor groove AT selectivity with oligonucleotides},
 type = {article},
 year = {1997},
 pages = {1623-1624},
 id = {9b3e2289-c499-39c6-9a25-43e57803b26c},
 created = {2023-01-10T01:46:56.245Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:56.245Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {NMR studies show that the ternary octahedral Δ-cis-α-[Ru(RR-picchxnMe2)(phen)]2+ cation binds with AT selectivity in the minor groove of [d(CGCGATCGCG)2] and [d(ATATCGATAT)2] duplexes through a non-intercalative interaction.},
 bibtype = {article},
 author = {Proudfoot, E.M. and Mackay, J.P. and Vagg, R.S. and Vickery, K.A. and Williams, P.A. and Karuso, P.},
 doi = {10.1039/a704001f},
 journal = {Chemical Communications},
 number = {17}
}

@article{
 title = {Assignment of the <sup>1</sup>H NMR spectrum and solution conformation of the antitumour antibiotic ditrisarubicin B},
 type = {article},
 year = {1996},
 pages = {5617-5624},
 volume = {52},
 id = {47129b2c-d23a-3485-8fba-32923cad98a3},
 created = {2023-01-10T01:46:57.152Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:57.152Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Complete assignment of the 1H NMR spectrum of ditrisarubicin B, a member of the anthracycline antitumour antibiotics, in acetonitrile is reported. The two trisaccharide chains are highly structured and their conformation supports their role as preorganised DNA minor groove binders which bind to DNA on intercalation of the tetracyclic chromophore.},
 bibtype = {article},
 author = {Mackay, J.P. and Shelton, C.J. and Harding, M.M.},
 doi = {10.1016/0040-4020(96)00198-6},
 journal = {Tetrahedron},
 number = {15}
}

@article{
 title = {Backbone dynamics of the c-Jun leucine zipper: <sup>15</sup>N NMR relaxation studies},
 type = {article},
 year = {1996},
 pages = {4867-4877},
 volume = {35},
 id = {362dbcd0-e82e-357a-a5d5-97ec3d2ac62e},
 created = {2023-01-10T01:46:58.102Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:58.102Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {The backbone dynamics of the coiled-coil leucine zipper domain of c-Jun have been studied using proton-detected two-dimensional 1H-15N NMR spectroscopy. Longitudinal (T1) and transverse (T2) 15N relaxation times, together with 1H15N NOEs, were measured and analyzed by considering the protein to approximate a prolate ellipsoid. An analysis of the T1/T2 ratios for residues in the well-structured section of the protein showed that a model for the spectral density function in which the protein is considered to reorient anisotropically fitted the data significantly better than an isotropic model. Order parameters (S2) in the range 0.7-0.9 were observed for most residues, with lower values near the C-terminus, consistent with fraying of the two helices comprising the coiled-coil. Because nearly all of the N-H vectors have small angles to the long axis of the molecule, there was some uncertainty in the value of the rotational diffusion coefficient Dpar, which describes rotation about the long axis. Thus, an alternative method was examined for its ability to provide independent estimates of Dpar and Dperp (the diffusion coefficient describing rotation about axes perpendicular to the long axis); the translational diffusion coefficient (Dt) of the protein was measured, and hydrodynamic calculations were used to predict Dpar and Dperp. However, the derived rotational diffusion coefficients proved to be very dependent on the hydrodynamic model used to relate Dt to Dpar and Dperp, and consequently the values obtained from the T1/T2 analysis were used in the order-parameter analysis. Although it has previously been reported that the side chain of a polar residue at the dimer interface, Asn22, undergoes a conformational exchange process and destabilizes the dimer, no evidence of increased backbone mobility in this region was detected, suggesting that this process is confined to the Asn side chain.},
 bibtype = {article},
 author = {Mackay, J.P. and Shaw, G.L. and King, G.F.},
 doi = {10.1021/bi952761y},
 journal = {Biochemistry},
 number = {15}
}

@article{
 title = {Development of a sensitive peptide-based immunoassay: Application to detection of the Jun and Fos oncoproteins},
 type = {article},
 year = {1996},
 pages = {9069-9075},
 volume = {35},
 id = {3ec35428-2577-35f5-8d93-7aae09b3b3bc},
 created = {2023-01-10T01:46:59.013Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:46:59.013Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {c-Jun and c-Fos belong to the bZIP class of transcriptional activator proteins, many of which have been implicated in the neoplastic transformation of cells. We are interested in engineering dominant-negative leucine zipper (LZ) peptides as a means of sequestering these proteins in vivo in order to suppress their transcriptional regulatory activity. Toward this end, we have developed a novel immunoassay for measuring the dimerization affinities of dimeric Jun and Fos complexes. This peptide-based ELISA relies on the fact that Jun and Fos preferentially form heterodimers via their leucine zipper domains. Recombinant Jun leucine zipper peptides (either native JunLZ or a V36→E point mutant) were labeled with biotin and specifically bound through a leucine zipper interaction to a FosLZ-glutathione S-transferase fusion protein adsorbed onto the wells of an ELISA tray. Jun:Fos complexes were subsequently detected using a recently developed streptavidin-based amplification system known as enzyme complex amplification [Wilson, M. R., and Easterbrook-Smith, S. B. (1993) Anal. Biochem. 209, 183-187]. This ELISA system can detect subnanomolar concentrations of Jun and Fos, thus allowing determination of the dissociation constants for complex formation. The dissociation constant for formation of the native JunLZ:FosLZ heterodimer at 37 °C was determined to be 0.99 ± 0.30 nM, while that for JunLZ(V36E):FosLZ heterodimer was 0.90 ± 0.13 μM. These results demonstrate that the novel peptide-based ELISA described herein is simple and sensitive and can be used to rapidly screen for potential dominant-negative leucine zipper peptides.},
 bibtype = {article},
 author = {Heuer, K.H. and Mackay, J.P. and Podzebenko, P. and Bains, N.P.S. and Weiss, A.S. and King, G.F. and Easterbrook-Smith, S.B.},
 doi = {10.1021/bi952817o},
 journal = {Biochemistry},
 number = {28}
}

@article{
 title = {Measuring protein self-association using pulsed-field-gradient NMR spectroscopy: Application to myosin light chain 2},
 type = {article},
 year = {1995},
 pages = {321-328},
 volume = {6},
 id = {a162714d-04d2-3f71-8080-eeb26343a53a},
 created = {2023-01-10T01:47:00.198Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:47:00.198Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {At the millimolar concentrations required for structural studies, NMR spectra of the calcium-binding protein myosin light chain 2 (MLC2) showed resonance line widths indicative of extensive self-association. Pulsed-field-gradient (PFG) NMR spectroscopy was used to examine whether MLC2 aggregation could be prevented by the zwitterionic bile salt derivative 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). PFG NMR measurements indicated that CHAPS was capable of preventing MLC2 self-association, but only at concentrations well above the critical micelle concentration of ∼7.5 mM. CHAPS was most effective at a concentration of 22.5 mM, where the apparent molecular mass of MLC2 correponded to a protein monomer plus seven molecules of bound detergent. The resolution and sensitivity of 2D 15N-1H HSQC spectra of MLC2 were markedly improved by the addition of 25 mM CHAPS, consistent with a reduction in aggregation following addition of the detergent. The average amide nitrogen T2 value for MLC2 increased from ∼30 ms in the absence of CHAPS to ∼56 ms in the presence of 25 mM CHAPS. The results of this study lead us to propose that PFG NMR spectroscopy can be used as a facile alternative to conventional techniques such as analytical ultracentrifugation for examining the self-association of biological macromolecules. © 1995 ESCOM Science Publishers B.V.},
 bibtype = {article},
 author = {Dingley, A.J. and Mackay, J.P. and Chapman, B.E. and Morris, M.B. and Kuchel, P.W. and Hambly, B.D. and King, G.F.},
 doi = {10.1007/BF00197813},
 journal = {Journal of Biomolecular NMR},
 number = {3}
}

@article{
 title = {Functional roles of natural products: The involvement of extended arrays of weak interactions in cooperative binding phenomena},
 type = {article},
 year = {1994},
 pages = {1975-1982},
 volume = {66},
 id = {c10c940c-9f14-3cf0-888e-39c896da615c},
 created = {2023-01-10T01:47:01.128Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:47:01.128Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {A factorisation of the free energy of binding into various “costs” and “benefits” has provided the basis for a semi-quantitation of some weak interactions in solution. It has become clear that the entropie cost of motional restriction on binding (A + B → A.B) increases with increasing exothermicity of the association; this exothermicity must of course reflect not only interactions at the interface between A and B, but also the change in bonding throughout B (if B represents the receptor). We illustrate cooperativity and anti-cooperativity by reference to effects of ligand binding (as models of classical agonists and antagonists) on the dimerisation of vancomycin-group antibiotics. Since dimerization of receptors (promoted by ligand binding) is a common theme in biological signalling, it must presumably have an advantage in natural selection. We suggest that concurrent demands of ligand binding and receptor dimerization may permit a more specific control of those ligand structures which can cause signal transmission. In this way, specificity in biological signalling might be aided. © 1994 IUPAC},
 bibtype = {article},
 author = {Williams, D.H. and Searle, M.S. and Groves, P. and Mackay, J.P. and Westwell, M.S. and Beauregard, D.A. and Cristofaro, M.F.},
 doi = {10.1351/pac199466101975},
 journal = {Pure and Applied Chemistry},
 number = {10-11}
}

@article{
 title = {Dissection of the Contributions toward Dimerization of Glycopeptide Antibiotics},
 type = {article},
 year = {1994},
 pages = {4573-4580},
 volume = {116},
 id = {5793ec9e-c2d9-3eba-9500-f54fc95f52df},
 created = {2023-01-10T01:47:02.045Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:47:02.045Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {A procedure for the determination of association constants in aqueous solution using hydrogen-deuterium exchange has been developed and used to measure the dimerization constant, Kam, for a number of strongly dimerizing glycopeptide antibiotics. These values provide further insight into the thermodynamic contributions of various structural epitopes to the dimerization of these antibiotics. Consideration of ligand binding affinities together with dimerization potentials provides evidence that dimerization is implicated in the physiological mode of action of these antibiotics. © 1994, American Chemical Society. All rights reserved.},
 bibtype = {article},
 author = {Mackay, J.P. and Gerhard, U. and Beauregard, D.A. and Maplestone, R.A. and Williams, D.H.},
 doi = {10.1021/ja00090a005},
 journal = {Journal of the American Chemical Society},
 number = {11}
}

@article{
 title = {Glycopeptide Antibiotic Activity and the Possible Role of Dimerization: A Model for Biological Signaling},
 type = {article},
 year = {1994},
 pages = {4581-4590},
 volume = {116},
 id = {71f945c5-5554-3649-bfe7-e37debf946b9},
 created = {2023-01-10T01:47:02.960Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:47:02.960Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {It is demonstrated that the presence of bacterial cell wall analogues may either enhance or, in the case of ristocetin A, oppose dimerization of glycopeptide antibiotics. These observations may imply that dimerization plays a role in the mode of action of these antibiotics, and a mechanism is proposed to take account of this possibility. The glycopeptide dimers are also found to be formed more exothermically in the presence of cell wall analogues, and the nature of biological signaling events is discussed in this context. It is pointed out that binding enthalpy (rather than simply binding free energy, ΔG) may be an important quantity in signaling events. If this is so, then oligomers may be abundant in signaling processes partly because the extended aggregates they form are able to cooperatively amplify the conformational changes which are incurred on ligand binding, which occur through relatively small changes in free energy but larger opposing changes in enthalpy and entropy. © 1994, American Chemical Society. All rights reserved.},
 bibtype = {article},
 author = {Mackay, J.P. and Gerhard, U. and Beauregard, D.A. and Westwell, M.S. and Searle, M.S. and Williams, D.H.},
 doi = {10.1021/ja00090a006},
 journal = {Journal of the American Chemical Society},
 number = {11}
}

@article{
 title = {The structure of an asymmetric dimer relevant to the mode of action of the glycopeptide antibiotics},
 type = {article},
 year = {1994},
 pages = {747-754},
 volume = {2},
 id = {19ebea8a-a41d-36ba-9081-a4f636b1f554},
 created = {2023-01-10T01:47:03.895Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:47:03.895Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Background Glycopeptide antibiotics of the vancomycin group are of crucial clinical importance in the treatment of methicillin resistant Staphylococcus aureus (MRSA) - the often lethal 'super-bug ' - characterized by its resistance to a wide range of antibiotics in common use. The antibiotics exert their physiological action by blocking cell wall synthesis through recognition of nascent cell wall mucopeptides terminating in the sequence -D-Ala-D -Ala. Evidence suggests that the antibiotics are able to enhance their biological activity by the formation of homodimers, and this is supported by the observation that dimerization and peptide binding in vitro are cooperative phenomena. The basis of this enhancement is not understood at the molecular level. Results The first detailed structure of a dimeric glycopeptide antibiotic, that of eremomycin, is presented based upon solution NMR data. The overall structure of the dimer complex is asymmetric. The source of this asymmetry - a parallel alignment and mutual interaction of the disaccharides - appears to promote dimerization through specific sugar- sugar recognition. Conclusions A molecular basis for the observed cooperativity of cell wall peptide binding by eremomycin is evident from these studies of the dimer. The carboxylate anion of the cell wall component, which is crucial to binding, forms an amide-mediated ion-pair interaction to the alkylammonium ion of the ring 6 sugar in the other half of the dimer making the structure and positioning of this sugar important in mediating cooperativity. © 1994 Elsevier Science Ltd. All rights reserved.},
 bibtype = {article},
 author = {Groves, P. and Searle, M.S. and Mackay, J.P. and Williams, D.H.},
 doi = {10.1016/S0969-2126(94)00075-1},
 journal = {Structure},
 number = {8}
}

@article{
 title = {The Role of the Sugar and Chlorine Substituents in the Dimerization of Vancomycin Antibiotics},
 type = {article},
 year = {1993},
 pages = {232-237},
 volume = {115},
 id = {26477609-41c1-3bb1-b211-10fe3f017959},
 created = {2023-01-10T01:47:04.826Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:47:04.826Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {Evidence is presented for the formation of dimers in aqueous solutions of the glycopeptide antibiotics eremomycin, A82846B, vancomycin, and eremomycin-ψ. The dimerization constant Kdim is determined by 1H NMR spectroscopy for the last two compounds and also for the related compound ristocetin-ψ, for which dimerization has previously been reported in mixed solvents. Values of Kdim are obtained for these compounds over a range of temperatures, and thus ΔHdim and ΔSdim are calculated. In addition, a lower limit for Kdim in the case of eremomycin is calculated (105 M−1). This is a remarkably large value, and it may be that dimerization is implicated in antibiotic action. The possibility that natural selection has led to adaptations which promote dimerization (such as the nature and sites of attachment of the sugars and a ring 2 chlorine atom) is discussed. © 1993, American Chemical Society. All rights reserved.},
 bibtype = {article},
 author = {Gerhard, U. and Mackay, J.P. and Maplestone, R.A. and Williams, D.H.},
 doi = {10.1021/ja00054a033},
 journal = {Journal of the American Chemical Society},
 number = {1}
}

@article{
 title = {Toward an estimation of binding constants in aqueous solution: Studies of associations of vancomycin group antibiotics},
 type = {article},
 year = {1993},
 pages = {1172-1178},
 volume = {90},
 id = {6e99bb91-e44e-3b3e-a076-022d9dbbba8a},
 created = {2023-01-10T01:47:05.775Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2023-01-10T01:47:05.775Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 private_publication = {false},
 abstract = {An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.},
 bibtype = {article},
 author = {Williams, D.H. and Searle, M.S. and Mackay, J.P. and Gerhard, U. and Maplestone, R.A.},
 doi = {10.1073/pnas.90.4.1172},
 journal = {Proceedings of the National Academy of Sciences of the United States of America},
 number = {4}
}

@article{
 title = {Macrocyclic peptides as a new class of targeted protein degraders},
 type = {article},
 year = {2025},
 pages = {326-337},
 volume = {6},
 id = {30eeaf46-43ed-3d7a-9d00-edc3ea4d8b2e},
 created = {2025-05-31T09:41:34.856Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:34.856Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Jing, X and Mackay, J P and Passioura, T},
 doi = {10.1039/D4CB00199K},
 journal = {RSC Chem Biol},
 number = {3}
}

@article{
 title = {Rapid Elaboration of Fragments into Leads applied to BRD3-extra terminal domain},
 type = {article},
 year = {2023},
 pages = {5859-5872},
 volume = {66},
 id = {8a395652-12ac-3c77-ab3f-a26469271e65},
 created = {2025-05-31T09:41:35.965Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:35.965Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Adams, L A and Wilkinson-White, L E and Gunzburg, M J and Headey, S J and Mohanty, B and Scanlon, M J and Capuano, B and Mackay, J P and Doak, B C},
 doi = {https://doi.org/10.1021/acs.jmedchem.3c00137},
 journal = {Journal of Medicinal Chemistry},
 number = {8}
}

@article{
 title = {Cross-linking mass spectrometry discovers, evaluates, and validates the experimental and predicted structural proteome},
 type = {article},
 year = {2023},
 pages = {e2219418120},
 volume = {120},
 id = {9ef702f4-96aa-3165-9804-08012f491b9c},
 created = {2025-05-31T09:41:37.182Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:37.182Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Bartolec, T K and Vazquez-Campos, X and Norman, A and Luong, C and Johnson, M and Payne, R J and Wilkins, M R and Mackay, J P and Low, J K K},
 doi = {10.1073/pnas.2219418120},
 journal = {Proceedings of the National Academy of Sciences, USA},
 number = {17}
}

@article{
 title = {Discovery and characterization of cyclic peptides selective for the C-terminal bromodomains of BET family proteins},
 type = {article},
 year = {2023},
 pages = {912-923},
 volume = {31},
 id = {648b9068-ebfb-3346-8d27-9d45c727b70a},
 created = {2025-05-31T09:41:38.442Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:38.442Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Franck, C and Patel, K and Walport, L J and Christie, M and Norman, A and Passioura, T and Suga, H and Payne, R J and Mackay, J P},
 journal = {Structure},
 number = {8}
}

@article{
 title = {The H2A.Z.1/PWWP2A/NuRD-associated protein HMG20A controls early head and heart developmental transcription programs},
 type = {article},
 year = {2023},
 pages = {472},
 volume = {14},
 id = {48231f79-8372-353d-b59c-8e93cd741d78},
 created = {2025-05-31T09:41:39.559Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:39.559Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Herchenrother, A and Gossen, S and Friedrich, T and Reim, A and Daus, N and Diegmuller, F and Sgellmacher, I and Szymkowiak, L and Nist, A and Stiewe, T and Borggrefe, T and Mann, M and Mackay, J P and Bartkuhn, M and Borchers, C H and Lan, J and Hake, S},
 journal = {Nature Commun}
}

@article{
 title = {DUF2285 is a novel helix-turn-helix domain variant that orchestrates both activation and antiactivation of conjugative element transfer in proteobacteria},
 type = {article},
 year = {2023},
 pages = {6841-6856},
 volume = {51},
 id = {cbf4a57e-6ac6-38ba-9663-ebbd12bff75a},
 created = {2025-05-31T09:41:40.678Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:40.678Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Jowsey, W J and Morris, C R P and Hall, D and Sullivan, J T and Fagerlund, R and Eto, K Y and Solomon, P D and Mackay, J P and Bond, C S and Ramsay, J P and Ronson, C W},
 journal = {Nucl Acids Res},
 number = {13}
}

@article{
 title = {Pathogenic human variant that dislocates GATA2 zinc fingers establishes hematopoiesis-disrupting gene expression and signaling networks},
 type = {article},
 year = {2023},
 pages = {e162685},
 volume = {133},
 id = {2b3b5924-1e5f-320a-adb4-f63ad5c8514c},
 created = {2025-05-31T09:41:41.809Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:41.809Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Jung, M M and Shen, S and Botten, G and Olender, T and Katsumura, K R and Johnson, K D and Soukup, A A and Liu, P and Zhang, Q and Jensfold, Z and Lewis, P W and Beagrie, R A and Low, J K K and Yang, L and Mackay, J P and Godley, J A and Brand, M and Xu, J and Keles, S and Bresnick, E H},
 journal = {Journal of Clinical Investigation},
 number = {7}
}

@article{
 title = {Fibroblast-expressed LRRC15 is a receptor for SARS-CoV-2 spike and controls antiviral and antifibrotic transcriptional programs},
 type = {article},
 year = {2023},
 pages = {e3001967},
 volume = {21},
 id = {963b0ec7-22a6-3509-8b48-23463178bccb},
 created = {2025-05-31T09:41:42.920Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:42.920Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Loo, L and Waller, M and Moreno, C and Cole, A and Ospina Stella, A and Pop, O-T and Jochum, A-K and Ali, O H and Denes, C and Hamoudi, Z and Chung, F and Aggarwal, A and Low, J K K and Patel, K and Siddiquee, R and Kang, T and Mathivanan, S and Mackay, J P and Jochum, W and Flatz, L and Hesselson, D and Turville, S G and Neely, G G},
 journal = {PLoS Biology},
 number = {2}
}

@article{
 title = {mRNA display reveals a class of high-affinity bromodomain-binding motifs that are not found in the human proteome},
 type = {article},
 year = {2023},
 pages = {105482-105495},
 volume = {299},
 id = {8138462d-5db6-3c2b-b928-0fc2e79158ae},
 created = {2025-05-31T09:41:44.027Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:44.027Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Low, J K K and Patel, K and Jones, N and Solomon, P and Norman, A and Maxwell, J W C and Pachl, P and Matthews, J M and Payne, R J and Passioura, T and Suga, H and Walport, L J and Mackay, J P},
 journal = {Journal of Biological Chemistry},
 number = {12}
}

@article{
 title = {A NuRD for all seasons},
 type = {article},
 year = {2023},
 pages = {11-25},
 volume = {48},
 id = {4bc6fc94-f576-3a43-8c67-cc27c02c71e4},
 created = {2025-05-31T09:41:45.132Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:45.132Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Reid, X J and Low, J K K and Mackay, J P},
 doi = {10.1016/j.tibs.2022.06.002},
 journal = {Trends Biochem Sci},
 number = {1}
}

@article{
 title = {De novo variants in GATAD2A in individuals with developmental disorders},
 type = {article},
 year = {2023},
 pages = {100198},
 volume = {4},
 id = {32a23d36-31d5-311d-9a62-efb778e063ed},
 created = {2025-05-31T09:41:46.229Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:46.229Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Werren, E A and Guxholli, A and Jones, N and Wagner, M and Hannibal, I and Granadillo, J L and Tyndall, A V and Moccia, A and Kuehl, R and Levandoski, K M and Day-Salvatore, D L and Wheeler, M and Genomics, University of Washington Center for Mendelian and Chong, J X and Bamshad, M J and Innes, M and Pierson, T M and Mackay, J P and Bielas, S L and Martin, D M},
 journal = {Human Genet Genom Adv},
 number = {3}
}

@article{
 title = {Discovery of High Affinity Cyclic Peptide Ligands for Human ACE2 with SARS-CoV-2 Entry Inhibitory Activity},
 type = {article},
 year = {2024},
 pages = {141-152},
 volume = {19},
 id = {4696afdc-c968-372b-a402-8946a161aaca},
 created = {2025-05-31T09:41:47.301Z},
 file_attached = {false},
 profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},
 group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},
 last_modified = {2025-05-31T09:41:47.301Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {false},
 hidden = {false},
 source_type = {Journal Article},
 private_publication = {false},
 bibtype = {article},
 author = {Bedding, M J and Franck, C and Johansen-Leete, J and Aggarwal, A and Maxwell, J W C and Patel, K and Hawkins, P and Low, J K K and Siddiquee, R and Moghaddas Sani, H and Ford, D J and Turville, S G and Mackay, J P and Passioura, T and Christie, M and Payne, R J},
 doi = {10.1021/acschembio.3c00568},
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 title = {ZNF827 is a single-stranded DNA binding protein that interacts with TOPBP1 to regulate the ATR-CHK1 DNA damage response pathway},
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 year = {2024},
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}\">\n            <i class=\"fa fa-download fa-fw\"></i> Download BibTeX\n          </a>\n        </li>\n        <li>\n          <a href=\"//bibbase.org/rss?bib=\">\n            <i class=\"fa fa-rss fa-fw\"></i> RSS Feed\n          </a>\n          </li>\n      </ul>\n      </li>\n\n      \n\n\n      \n    </ul>\n\n\n    <div class=\"alert alert-success bibbase_msg\" id=\"bibbase_msg_embed\"\n         style=\"display: none;\">\n\n      Excellent! 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In order to keep using Mendeley with BibBase past April\n        14th, you need to:\n        <ol>\n          <li>renew the authorization for BibBase on Mendeley, and</li>\n          <li>update the BibBase URL\n            in your page the same way you did when you initially set up\n            this page.\n          </li>\n        </ol>\n      </p>\n\n      <p>\n        <a href=\"http://bibbase.org/cgi-bin/mendeley2/get.py\">\n          <i class=\"fa fa-angle-double-right\"></i>\n          Fix it now</a>\n      </p>\n    </div>\n\n  </div>\n\n\n  <div id=\"bibbase_body\">\n    \n  \n  <div class=\"bibbase_group_whole\" id=\"group_2025\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2025')\"\n      onkeypress=\"toggleGroup('group_2025')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2025</span>\n      <span class=\"bibbase_group_count\">\n        \n        (1)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"jing-mackay-passioura-macrocyclicpeptidesasanewclassoftargetedproteindegraders-2025\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Macrocyclic peptides as a new class of targeted protein degraders.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Jing,  X.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Passioura,  T.\n</span>\n\n  \n\n</span>\n\n  <i>RSC Chem Biol</i>, 6(3): 326-337.  2025.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1039/D4CB00199K\"\n     onclick=\"log_download('jing-mackay-passioura-macrocyclicpeptidesasanewclassoftargetedproteindegraders-2025', 'http://doi.org/10.1039/D4CB00199K', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/jing-mackay-passioura-macrocyclicpeptidesasanewclassoftargetedproteindegraders-2025\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('jing-mackay-passioura-macrocyclicpeptidesasanewclassoftargetedproteindegraders-2025')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Macrocyclic peptides as a new class of targeted protein degraders},\n type = {article},\n year = {2025},\n pages = {326-337},\n volume = {6},\n id = {30eeaf46-43ed-3d7a-9d00-edc3ea4d8b2e},\n created = {2025-05-31T09:41:34.856Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:34.856Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Jing, X and Mackay, J P and Passioura, T},\n doi = {10.1039/D4CB00199K},\n journal = {RSC Chem Biol},\n number = {3}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2024\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2024')\"\n      onkeypress=\"toggleGroup('group_2024')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2024</span>\n      <span class=\"bibbase_group_count\">\n        \n        (11)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"bedding-franck-johansenleete-aggarwal-maxwell-patel-hawkins-low-etal-discoveryofhighaffinitycyclicpeptideligandsforhumanace2withsarscov2entryinhibitoryactivity-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Discovery of High Affinity Cyclic Peptide Ligands for Human ACE2 with SARS-CoV-2 Entry Inhibitory Activity.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Bedding,  M., J.; Franck,  C.; Johansen-Leete,  J.; Aggarwal,  A.; Maxwell,  J., W., C.; Patel,  K.; Hawkins,  P.; Low,  J., K., K.; Siddiquee,  R.; Moghaddas Sani,  H.; Ford,  D., J.; Turville,  S., G.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Passioura,  T.; Christie,  M.;  and Payne,  R., J.\n</span>\n\n  \n\n</span>\n\n  <i>ACS Chemical Biology</i>, 19: 141-152.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/acschembio.3c00568\"\n     onclick=\"log_download('bedding-franck-johansenleete-aggarwal-maxwell-patel-hawkins-low-etal-discoveryofhighaffinitycyclicpeptideligandsforhumanace2withsarscov2entryinhibitoryactivity-2024', 'http://doi.org/10.1021/acschembio.3c00568', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/bedding-franck-johansenleete-aggarwal-maxwell-patel-hawkins-low-etal-discoveryofhighaffinitycyclicpeptideligandsforhumanace2withsarscov2entryinhibitoryactivity-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('bedding-franck-johansenleete-aggarwal-maxwell-patel-hawkins-low-etal-discoveryofhighaffinitycyclicpeptideligandsforhumanace2withsarscov2entryinhibitoryactivity-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Discovery of High Affinity Cyclic Peptide Ligands for Human ACE2 with SARS-CoV-2 Entry Inhibitory Activity},\n type = {article},\n year = {2024},\n pages = {141-152},\n volume = {19},\n id = {4696afdc-c968-372b-a402-8946a161aaca},\n created = {2025-05-31T09:41:47.301Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:47.301Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Bedding, M J and Franck, C and Johansen-Leete, J and Aggarwal, A and Maxwell, J W C and Patel, K and Hawkins, P and Low, J K K and Siddiquee, R and Moghaddas Sani, H and Ford, D J and Turville, S G and Mackay, J P and Passioura, T and Christie, M and Payne, R J},\n doi = {10.1021/acschembio.3c00568},\n journal = {ACS Chemical Biology}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"dilly-franck-bhusal-morgan-bedding-low-mackay-stone-etal-tyrosinesulfationmodulatesthebindingactivityofchemokinetargetingnanobodies-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Tyrosine Sulfation Modulates the Binding Activity of Chemokine-Targeting Nanobodies.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dilly,  J., J.; Franck,  C.; Bhusal,  R.; Morgan,  A.; Bedding,  M., J.; Low,  J., K., K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Stone,  M., J.;  and Payne,  R., J.\n</span>\n\n  \n\n</span>\n\n  <i>ACS Chemical Biology</i>, 19(7): 1426-1432.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dilly-franck-bhusal-morgan-bedding-low-mackay-stone-etal-tyrosinesulfationmodulatesthebindingactivityofchemokinetargetingnanobodies-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dilly-franck-bhusal-morgan-bedding-low-mackay-stone-etal-tyrosinesulfationmodulatesthebindingactivityofchemokinetargetingnanobodies-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Tyrosine Sulfation Modulates the Binding Activity of Chemokine-Targeting Nanobodies},\n type = {article},\n year = {2024},\n pages = {1426-1432},\n volume = {19},\n id = {66562371-6596-3a35-a0d3-6e4f7cdec103},\n created = {2025-05-31T09:41:48.446Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:48.446Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Dilly, J J and Franck, C and Bhusal, R and Morgan, A and Bedding, M J and Low, J K K and Mackay, J P and Stone, M J and Payne, R J},\n journal = {ACS Chemical Biology},\n number = {7}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"du-hall-chung-kurdyukov-crittenden-patel-dawson-westhorpe-etal-moleculardissectionofcobravenomhighlightsheparinoidsasaneffectivesnakebiteantidote-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Molecular dissection of cobra venom highlights heparinoids as an effective snakebite antidote.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Du,  T., Y.; Hall,  S., R.; Chung,  F.; Kurdyukov,  S.; Crittenden,  E.; Patel,  K.; Dawson,  C., A.; Westhorpe,  A., P.; Bartlett,  K., E.; Rasmussen,  S., A.; Moreno,  C.; Denes,  C., E.; Albulescu,  L.; Marriott,  A., E.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Wilkinson,  M., C.; Gutierrez,  J., M.; Casewell,  N., R.;  and Neely,  G., G.\n</span>\n\n  \n\n</span>\n\n  <i>Science Transl. Med.</i>, 16: eadk4802.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/du-hall-chung-kurdyukov-crittenden-patel-dawson-westhorpe-etal-moleculardissectionofcobravenomhighlightsheparinoidsasaneffectivesnakebiteantidote-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('du-hall-chung-kurdyukov-crittenden-patel-dawson-westhorpe-etal-moleculardissectionofcobravenomhighlightsheparinoidsasaneffectivesnakebiteantidote-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Molecular dissection of cobra venom highlights heparinoids as an effective snakebite antidote},\n type = {article},\n year = {2024},\n pages = {eadk4802},\n volume = {16},\n id = {b64633c3-ede3-300e-a1e1-178673c2746c},\n created = {2025-05-31T09:41:49.759Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:49.759Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Du, T Y and Hall, S R and Chung, F and Kurdyukov, S and Crittenden, E and Patel, K and Dawson, C A and Westhorpe, A P and Bartlett, K E and Rasmussen, S A and Moreno, C and Denes, C E and Albulescu, L-O and Marriott, A E and Mackay, J P and Wilkinson, M C and Gutierrez, J M and Casewell, N R and Neely, G G},\n journal = {Science Transl. Med.}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"imoto-xue-luo-raychaudhuri-itoh-ma-craft-kwan-etal-dynamin1xainteractswithendophilina1viaitssplicedlongcterminusforultrafastendocytosis-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Dynamin 1xA interacts with Endophilin A1 via its spliced long C-terminus for ultrafast endocytosis.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Imoto,  Y.; Xue,  J.; Luo,  L.; Raychaudhuri,  S.; Itoh,  K.; Ma,  Y.; Craft,  G., E.; Kwan,  A., H.; Ogunmowo,  T., H.; Ho,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Ha,  T.; Watanabe,  S.;  and Robinson,  P., J.\n</span>\n\n  \n\n</span>\n\n  <i>EMBO Journal</i>, 43(16): 3327-3357.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s44318-024-00145-x\"\n     onclick=\"log_download('imoto-xue-luo-raychaudhuri-itoh-ma-craft-kwan-etal-dynamin1xainteractswithendophilina1viaitssplicedlongcterminusforultrafastendocytosis-2024', 'http://doi.org/10.1038/s44318-024-00145-x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/imoto-xue-luo-raychaudhuri-itoh-ma-craft-kwan-etal-dynamin1xainteractswithendophilina1viaitssplicedlongcterminusforultrafastendocytosis-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('imoto-xue-luo-raychaudhuri-itoh-ma-craft-kwan-etal-dynamin1xainteractswithendophilina1viaitssplicedlongcterminusforultrafastendocytosis-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Dynamin 1xA interacts with Endophilin A1 via its spliced long C-terminus for ultrafast endocytosis},\n type = {article},\n year = {2024},\n pages = {3327-3357},\n volume = {43},\n id = {7a93a949-e534-3a31-a560-479c1efe8953},\n created = {2025-05-31T09:41:51.012Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:51.012Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Imoto, Y and Xue, J and Luo, L and Raychaudhuri, S and Itoh, K and Ma, Y and Craft, G E and Kwan, A H and Ogunmowo, T H and Ho, A and Mackay, J P and Ha, T and Watanabe, S and Robinson, P J},\n doi = {10.1038/s44318-024-00145-x},\n journal = {EMBO Journal},\n number = {16}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kambanis-ayoub-bedding-egelund-maxwell-franck-lambrechts-hawkins-etal-expressedproteinligationinflow-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Expressed protein ligation in flow.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kambanis,  L.; Ayoub,  A.; Bedding,  M., J.; Egelund,  P., H., G.; Maxwell,  J., W., C.; Franck,  C.; Lambrechts,  L.; Hawkins,  P., M., E.; Chisholm,  T., S.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Sierecki,  E.; Gambin,  Y.; Kulkarni,  S., S.;  and Payne,  R., J.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of the American Chemical Society</i>, 146(31): 22027-22035.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kambanis-ayoub-bedding-egelund-maxwell-franck-lambrechts-hawkins-etal-expressedproteinligationinflow-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kambanis-ayoub-bedding-egelund-maxwell-franck-lambrechts-hawkins-etal-expressedproteinligationinflow-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Expressed protein ligation in flow},\n type = {article},\n year = {2024},\n pages = {22027-22035},\n volume = {146},\n id = {0b1e7050-2eb0-370b-bb29-430cfb06aad1},\n created = {2025-05-31T09:41:52.471Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:52.471Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Kambanis, L and Ayoub, A and Bedding, M J and Egelund, P H G and Maxwell, J W C and Franck, C and Lambrechts, L and Hawkins, P M E and Chisholm, T S and Mackay, J P and Sierecki, E and Gambin, Y and Kulkarni, S S and Payne, R J},\n journal = {Journal of the American Chemical Society},\n number = {31}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kyrilis-low-mackay-panagiotis-structuralbiologyincellulomindingthegapbetweenconceptualizationandrealization-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural Biology in cellulo: Minding the gap between conceptualization and realization.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kyrilis,  F.; Low,  J., K., K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Panagiotis,  K.\n</span>\n\n  \n\n</span>\n\n  <i>Current Opinion in Structural Biology</i>, 87: 102843.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kyrilis-low-mackay-panagiotis-structuralbiologyincellulomindingthegapbetweenconceptualizationandrealization-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kyrilis-low-mackay-panagiotis-structuralbiologyincellulomindingthegapbetweenconceptualizationandrealization-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural Biology in cellulo: Minding the gap between conceptualization and realization},\n type = {article},\n year = {2024},\n pages = {102843},\n volume = {87},\n id = {442e4df5-4a08-3fc8-a4aa-a0a2f78862ec},\n created = {2025-05-31T09:41:53.718Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:53.718Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Kyrilis, F and Low, J K K and Mackay, J P and Panagiotis, K},\n journal = {Current Opinion in Structural Biology}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"murray-valli-milazzo-mayoh-gifford-fletcher-xue-jayatilleke-etal-thetranscriptionalcorepressorrunx1t1isessentialfornmycdrivenneuroblastomatumorigenesis-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The transcriptional co-repressor Runx1t1 is essential for N-Myc-driven neuroblastoma tumorigenesis.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Murray,  J., E.; Valli,  E.; Milazzo,  G.; Mayoh,  C.; Gifford,  A., J.; Fletcher,  J., I.; Xue,  C.; Jayatilleke,  N.; Salehzadeh,  F.; Gamble,  L.; Rouaen,  J.; Carter,  D.; Forgham,  H.; Sekyere,  E., O.; Keating,  J.; Eden,  G.; Allan,  S.; Alfred,  S.; Kusuma,  F.; Clark,  A.; Webber,  H.; Russell,  A., J.; de Weck,  A.; Kile,  B., T.; Santulli,  M.; De Rosa,  P.; Fleuren,  E., D., G.; Gao,  W.; Wilkinson-White,  L.; Low,  J., K., K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Marshall,  G., M.; Hilton,  D., J.; Giorgi,  F.; Koster,  J.; Perini,  G.; Haber,  M.;  and Norris,  M., D.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Commun</i>, 15(1): 5585.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/murray-valli-milazzo-mayoh-gifford-fletcher-xue-jayatilleke-etal-thetranscriptionalcorepressorrunx1t1isessentialfornmycdrivenneuroblastomatumorigenesis-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('murray-valli-milazzo-mayoh-gifford-fletcher-xue-jayatilleke-etal-thetranscriptionalcorepressorrunx1t1isessentialfornmycdrivenneuroblastomatumorigenesis-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The transcriptional co-repressor Runx1t1 is essential for N-Myc-driven neuroblastoma tumorigenesis},\n type = {article},\n year = {2024},\n pages = {5585},\n volume = {15},\n id = {68dab786-beb0-3d2d-865d-aae75e26b4d3},\n created = {2025-05-31T09:41:54.902Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:54.902Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Murray, J E and Valli, E and Milazzo, G and Mayoh, C and Gifford, A J and Fletcher, J I and Xue, C and Jayatilleke, N and Salehzadeh, F and Gamble, L and Rouaen, J and Carter, D and Forgham, H and Sekyere, E O and Keating, J and Eden, G and Allan, S and Alfred, S and Kusuma, F and Clark, A and Webber, H and Russell, A J and de Weck, A and Kile, B T and Santulli, M and De Rosa, P and Fleuren, E D G and Gao, W and Wilkinson-White, L and Low, J K K and Mackay, J P and Marshall, G M and Hilton, D J and Giorgi, F and Koster, J and Perini, G and Haber, M and Norris, M D},\n journal = {Nature Commun},\n number = {1}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"rajendiran-devaraju-haddad-ravi-panigrahi-paul-gopalakrishnan-wyman-etal-baseeditingofkeyresiduesinthebcl11axlspecificzincfingerdomainsderepressesfetalglobinexpression-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Base editing of key residues in the BCL11A-XL-specific zinc finger domains derepresses fetal globin expression.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Rajendiran,  V.; Devaraju,  N.; Haddad,  M.; Ravi,  N., S.; Panigrahi,  L.; Paul,  J.; Gopalakrishnan,  C.; Wyman,  S.; Ariudainambi,  K.; Mahalingam,  G.; Perisasami,  Y.; Prasad,  K.; George,  A.; Sukimaran,  D.; Gopinathan,  S.; Pai,  A., A.; Nakamura,  Y.; Balsubramanian,  P.; Ramalingam,  R.; Thangavel,  S.; Velayudhan,  S., R.; Corn,  J., E.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Marepally,  S.; Srivastava,  A.; Crossley,  M.;  and Mohankumar,  K., M.\n</span>\n\n  \n\n</span>\n\n  <i>Molecular Therapy</i>, 32: 663-677.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/rajendiran-devaraju-haddad-ravi-panigrahi-paul-gopalakrishnan-wyman-etal-baseeditingofkeyresiduesinthebcl11axlspecificzincfingerdomainsderepressesfetalglobinexpression-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('rajendiran-devaraju-haddad-ravi-panigrahi-paul-gopalakrishnan-wyman-etal-baseeditingofkeyresiduesinthebcl11axlspecificzincfingerdomainsderepressesfetalglobinexpression-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Base editing of key residues in the BCL11A-XL-specific zinc finger domains derepresses fetal globin expression},\n type = {article},\n year = {2024},\n pages = {663-677},\n volume = {32},\n id = {9d25591f-af39-35e5-8b3c-f698858d8d65},\n created = {2025-05-31T09:41:56.107Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:56.107Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Rajendiran, V and Devaraju, N and Haddad, M and Ravi, N S and Panigrahi, L and Paul, J and Gopalakrishnan, C and Wyman, S and Ariudainambi, K and Mahalingam, G and Perisasami, Y and Prasad, K and George, A and Sukimaran, D and Gopinathan, S and Pai, A A and Nakamura, Y and Balsubramanian, P and Ramalingam, R and Thangavel, S and Velayudhan, S R and Corn, J E and Mackay, J P and Marepally, S and Srivastava, A and Crossley, M and Mohankumar, K M},\n journal = {Molecular Therapy}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wunderlich-deshpande-paasche-friedrich-diegmuller-haddad-kreienbaum-naseer-etal-znf512bbindsrbbp4viaavariantnurdinteractionmotifandaggregateschromatininanurdcomplexindependentmanner-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  ZNF512B binds RBBP4 via a variant NuRD interaction motif and aggregates chromatin in a NuRD complex-independent manner.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wunderlich,  T.; Deshpande,  C.; Paasche,  L.; Friedrich,  T.; Diegmuller,  F.; Haddad,  E.; Kreienbaum,  C.; Naseer,  H.; Stebel,  S.; Daus,  N.; Leers,  J.; Lan,  J.; Trinh,  V.; Vazquez,  O.; Butter,  F.; Bartkuhn,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Hake,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Nucl Acids Res</i>, 52(21): 12831-12849.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wunderlich-deshpande-paasche-friedrich-diegmuller-haddad-kreienbaum-naseer-etal-znf512bbindsrbbp4viaavariantnurdinteractionmotifandaggregateschromatininanurdcomplexindependentmanner-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wunderlich-deshpande-paasche-friedrich-diegmuller-haddad-kreienbaum-naseer-etal-znf512bbindsrbbp4viaavariantnurdinteractionmotifandaggregateschromatininanurdcomplexindependentmanner-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {ZNF512B binds RBBP4 via a variant NuRD interaction motif and aggregates chromatin in a NuRD complex-independent manner},\n type = {article},\n year = {2024},\n pages = {12831-12849},\n volume = {52},\n id = {6f12feef-8089-3e58-aa15-f95cdeaaa0ec},\n created = {2025-05-31T09:41:57.310Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:57.310Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Wunderlich, T and Deshpande, C and Paasche, L and Friedrich, T and Diegmuller, F and Haddad, E and Kreienbaum, C and Naseer, H and Stebel, S and Daus, N and Leers, J and Lan, J and Trinh, V and Vazquez, O and Butter, F and Bartkuhn, M and Mackay, J P and Hake, S},\n journal = {Nucl Acids Res},\n number = {21}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"reid-zhong-mackay-howdoeschd4slidenucleosomes-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  How does CHD4 slide nucleosomes?.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Reid,  X., J.; Zhong,  Y.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Biochemical Society Transactions</i>, 52(5): 1995-2008.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1042/BST20230070\"\n     onclick=\"log_download('reid-zhong-mackay-howdoeschd4slidenucleosomes-2024', 'http://doi.org/10.1042/BST20230070', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/reid-zhong-mackay-howdoeschd4slidenucleosomes-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('reid-zhong-mackay-howdoeschd4slidenucleosomes-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {How does CHD4 slide nucleosomes?},\n type = {article},\n year = {2024},\n pages = {1995-2008},\n volume = {52},\n id = {b7e08595-1fa1-3d29-941a-320ad35365fe},\n created = {2025-05-31T09:41:58.530Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:58.530Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Reid, X J and Zhong, Y and Mackay, J P},\n doi = {10.1042/BST20230070},\n journal = {Biochemical Society Transactions},\n number = {5}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"yang-nelson-wells-fernando-lu-allen-malloy-lamm-etal-znf827isasinglestrandeddnabindingproteinthatinteractswithtopbp1toregulatetheatrchk1dnadamageresponsepathway-2024\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  ZNF827 is a single-stranded DNA binding protein that interacts with TOPBP1 to regulate the ATR-CHK1 DNA damage response pathway.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Yang,  S., F.; Nelson,  C., B.; Wells,  J., K.; Fernando,  M.; Lu,  R.; Allen,  R., A.; Malloy,  L.; Lamm,  N.; Murphy,  V., J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Deans,  A., J.; Cesare,  A., J.; Sobinoff,  A., P.;  and Pickett,  H., A.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Commun</i>, 15: 2210.  2024.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/yang-nelson-wells-fernando-lu-allen-malloy-lamm-etal-znf827isasinglestrandeddnabindingproteinthatinteractswithtopbp1toregulatetheatrchk1dnadamageresponsepathway-2024\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('yang-nelson-wells-fernando-lu-allen-malloy-lamm-etal-znf827isasinglestrandeddnabindingproteinthatinteractswithtopbp1toregulatetheatrchk1dnadamageresponsepathway-2024')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {ZNF827 is a single-stranded DNA binding protein that interacts with TOPBP1 to regulate the ATR-CHK1 DNA damage response pathway},\n type = {article},\n year = {2024},\n pages = {2210},\n volume = {15},\n id = {62819c41-9ebd-3ddf-9fc4-41f873dc1a24},\n created = {2025-05-31T09:41:59.693Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:59.693Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Yang, S F and Nelson, C B and Wells, J K and Fernando, M and Lu, R and Allen, R A and Malloy, L and Lamm, N and Murphy, V J and Mackay, J P and Deans, A J and Cesare, A J and Sobinoff, A P and Pickett, H A},\n journal = {Nature Commun}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2023\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2023')\"\n      onkeypress=\"toggleGroup('group_2023')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2023</span>\n      <span class=\"bibbase_group_count\">\n        \n        (10)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"adams-wilkinsonwhite-gunzburg-headey-mohanty-scanlon-capuano-mackay-etal-rapidelaborationoffragmentsintoleadsappliedtobrd3extraterminaldomain-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Rapid Elaboration of Fragments into Leads applied to BRD3-extra terminal domain.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Adams,  L., A.; Wilkinson-White,  L., E.; Gunzburg,  M., J.; Headey,  S., J.; Mohanty,  B.; Scanlon,  M., J.; Capuano,  B.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Doak,  B., C.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Medicinal Chemistry</i>, 66(8): 5859-5872.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/https://doi.org/10.1021/acs.jmedchem.3c00137\"\n     onclick=\"log_download('adams-wilkinsonwhite-gunzburg-headey-mohanty-scanlon-capuano-mackay-etal-rapidelaborationoffragmentsintoleadsappliedtobrd3extraterminaldomain-2023', 'http://doi.org/https://doi.org/10.1021/acs.jmedchem.3c00137', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/adams-wilkinsonwhite-gunzburg-headey-mohanty-scanlon-capuano-mackay-etal-rapidelaborationoffragmentsintoleadsappliedtobrd3extraterminaldomain-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('adams-wilkinsonwhite-gunzburg-headey-mohanty-scanlon-capuano-mackay-etal-rapidelaborationoffragmentsintoleadsappliedtobrd3extraterminaldomain-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Rapid Elaboration of Fragments into Leads applied to BRD3-extra terminal domain},\n type = {article},\n year = {2023},\n pages = {5859-5872},\n volume = {66},\n id = {8a395652-12ac-3c77-ab3f-a26469271e65},\n created = {2025-05-31T09:41:35.965Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:35.965Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Adams, L A and Wilkinson-White, L E and Gunzburg, M J and Headey, S J and Mohanty, B and Scanlon, M J and Capuano, B and Mackay, J P and Doak, B C},\n doi = {https://doi.org/10.1021/acs.jmedchem.3c00137},\n journal = {Journal of Medicinal Chemistry},\n number = {8}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"bartolec-vazquezcampos-norman-luong-johnson-payne-wilkins-mackay-etal-crosslinkingmassspectrometrydiscoversevaluatesandvalidatestheexperimentalandpredictedstructuralproteome-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Cross-linking mass spectrometry discovers, evaluates, and validates the experimental and predicted structural proteome.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Bartolec,  T., K.; Vazquez-Campos,  X.; Norman,  A.; Luong,  C.; Johnson,  M.; Payne,  R., J.; Wilkins,  M., R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Low,  J., K., K.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences, USA</i>, 120(17): e2219418120.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.2219418120\"\n     onclick=\"log_download('bartolec-vazquezcampos-norman-luong-johnson-payne-wilkins-mackay-etal-crosslinkingmassspectrometrydiscoversevaluatesandvalidatestheexperimentalandpredictedstructuralproteome-2023', 'http://doi.org/10.1073/pnas.2219418120', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/bartolec-vazquezcampos-norman-luong-johnson-payne-wilkins-mackay-etal-crosslinkingmassspectrometrydiscoversevaluatesandvalidatestheexperimentalandpredictedstructuralproteome-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('bartolec-vazquezcampos-norman-luong-johnson-payne-wilkins-mackay-etal-crosslinkingmassspectrometrydiscoversevaluatesandvalidatestheexperimentalandpredictedstructuralproteome-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Cross-linking mass spectrometry discovers, evaluates, and validates the experimental and predicted structural proteome},\n type = {article},\n year = {2023},\n pages = {e2219418120},\n volume = {120},\n id = {9ef702f4-96aa-3165-9804-08012f491b9c},\n created = {2025-05-31T09:41:37.182Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:37.182Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Bartolec, T K and Vazquez-Campos, X and Norman, A and Luong, C and Johnson, M and Payne, R J and Wilkins, M R and Mackay, J P and Low, J K K},\n doi = {10.1073/pnas.2219418120},\n journal = {Proceedings of the National Academy of Sciences, USA},\n number = {17}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"franck-patel-walport-christie-norman-passioura-suga-payne-etal-discoveryandcharacterizationofcyclicpeptidesselectiveforthecterminalbromodomainsofbetfamilyproteins-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Discovery and characterization of cyclic peptides selective for the C-terminal bromodomains of BET family proteins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Franck,  C.; Patel,  K.; Walport,  L., J.; Christie,  M.; Norman,  A.; Passioura,  T.; Suga,  H.; Payne,  R., J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 31(8): 912-923.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/franck-patel-walport-christie-norman-passioura-suga-payne-etal-discoveryandcharacterizationofcyclicpeptidesselectiveforthecterminalbromodomainsofbetfamilyproteins-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('franck-patel-walport-christie-norman-passioura-suga-payne-etal-discoveryandcharacterizationofcyclicpeptidesselectiveforthecterminalbromodomainsofbetfamilyproteins-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Discovery and characterization of cyclic peptides selective for the C-terminal bromodomains of BET family proteins},\n type = {article},\n year = {2023},\n pages = {912-923},\n volume = {31},\n id = {648b9068-ebfb-3346-8d27-9d45c727b70a},\n created = {2025-05-31T09:41:38.442Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:38.442Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Franck, C and Patel, K and Walport, L J and Christie, M and Norman, A and Passioura, T and Suga, H and Payne, R J and Mackay, J P},\n journal = {Structure},\n number = {8}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"herchenrother-gossen-friedrich-reim-daus-diegmuller-sgellmacher-szymkowiak-etal-theh2az1pwwp2anurdassociatedproteinhmg20acontrolsearlyheadandheartdevelopmentaltranscriptionprograms-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The H2A.Z.1/PWWP2A/NuRD-associated protein HMG20A controls early head and heart developmental transcription programs.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Herchenrother,  A.; Gossen,  S.; Friedrich,  T.; Reim,  A.; Daus,  N.; Diegmuller,  F.; Sgellmacher,  I.; Szymkowiak,  L.; Nist,  A.; Stiewe,  T.; Borggrefe,  T.; Mann,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Bartkuhn,  M.; Borchers,  C., H.; Lan,  J.;  and Hake,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Commun</i>, 14: 472.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/herchenrother-gossen-friedrich-reim-daus-diegmuller-sgellmacher-szymkowiak-etal-theh2az1pwwp2anurdassociatedproteinhmg20acontrolsearlyheadandheartdevelopmentaltranscriptionprograms-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('herchenrother-gossen-friedrich-reim-daus-diegmuller-sgellmacher-szymkowiak-etal-theh2az1pwwp2anurdassociatedproteinhmg20acontrolsearlyheadandheartdevelopmentaltranscriptionprograms-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The H2A.Z.1/PWWP2A/NuRD-associated protein HMG20A controls early head and heart developmental transcription programs},\n type = {article},\n year = {2023},\n pages = {472},\n volume = {14},\n id = {48231f79-8372-353d-b59c-8e93cd741d78},\n created = {2025-05-31T09:41:39.559Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:39.559Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Herchenrother, A and Gossen, S and Friedrich, T and Reim, A and Daus, N and Diegmuller, F and Sgellmacher, I and Szymkowiak, L and Nist, A and Stiewe, T and Borggrefe, T and Mann, M and Mackay, J P and Bartkuhn, M and Borchers, C H and Lan, J and Hake, S},\n journal = {Nature Commun}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"jowsey-morris-hall-sullivan-fagerlund-eto-solomon-mackay-etal-duf2285isanovelhelixturnhelixdomainvariantthatorchestratesbothactivationandantiactivationofconjugativeelementtransferinproteobacteria-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  DUF2285 is a novel helix-turn-helix domain variant that orchestrates both activation and antiactivation of conjugative element transfer in proteobacteria.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Jowsey,  W., J.; Morris,  C., R., P.; Hall,  D.; Sullivan,  J., T.; Fagerlund,  R.; Eto,  K., Y.; Solomon,  P., D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Bond,  C., S.; Ramsay,  J., P.;  and Ronson,  C., W.\n</span>\n\n  \n\n</span>\n\n  <i>Nucl Acids Res</i>, 51(13): 6841-6856.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/jowsey-morris-hall-sullivan-fagerlund-eto-solomon-mackay-etal-duf2285isanovelhelixturnhelixdomainvariantthatorchestratesbothactivationandantiactivationofconjugativeelementtransferinproteobacteria-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('jowsey-morris-hall-sullivan-fagerlund-eto-solomon-mackay-etal-duf2285isanovelhelixturnhelixdomainvariantthatorchestratesbothactivationandantiactivationofconjugativeelementtransferinproteobacteria-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {DUF2285 is a novel helix-turn-helix domain variant that orchestrates both activation and antiactivation of conjugative element transfer in proteobacteria},\n type = {article},\n year = {2023},\n pages = {6841-6856},\n volume = {51},\n id = {cbf4a57e-6ac6-38ba-9663-ebbd12bff75a},\n created = {2025-05-31T09:41:40.678Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:40.678Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Jowsey, W J and Morris, C R P and Hall, D and Sullivan, J T and Fagerlund, R and Eto, K Y and Solomon, P D and Mackay, J P and Bond, C S and Ramsay, J P and Ronson, C W},\n journal = {Nucl Acids Res},\n number = {13}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"jung-shen-botten-olender-katsumura-johnson-soukup-liu-etal-pathogenichumanvariantthatdislocatesgata2zincfingersestablisheshematopoiesisdisruptinggeneexpressionandsignalingnetworks-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Pathogenic human variant that dislocates GATA2 zinc fingers establishes hematopoiesis-disrupting gene expression and signaling networks.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Jung,  M., M.; Shen,  S.; Botten,  G.; Olender,  T.; Katsumura,  K., R.; Johnson,  K., D.; Soukup,  A., A.; Liu,  P.; Zhang,  Q.; Jensfold,  Z.; Lewis,  P., W.; Beagrie,  R., A.; Low,  J., K., K.; Yang,  L.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Godley,  J., A.; Brand,  M.; Xu,  J.; Keles,  S.;  and Bresnick,  E., H.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Clinical Investigation</i>, 133(7): e162685.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/jung-shen-botten-olender-katsumura-johnson-soukup-liu-etal-pathogenichumanvariantthatdislocatesgata2zincfingersestablisheshematopoiesisdisruptinggeneexpressionandsignalingnetworks-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('jung-shen-botten-olender-katsumura-johnson-soukup-liu-etal-pathogenichumanvariantthatdislocatesgata2zincfingersestablisheshematopoiesisdisruptinggeneexpressionandsignalingnetworks-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Pathogenic human variant that dislocates GATA2 zinc fingers establishes hematopoiesis-disrupting gene expression and signaling networks},\n type = {article},\n year = {2023},\n pages = {e162685},\n volume = {133},\n id = {2b3b5924-1e5f-320a-adb4-f63ad5c8514c},\n created = {2025-05-31T09:41:41.809Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:41.809Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Jung, M M and Shen, S and Botten, G and Olender, T and Katsumura, K R and Johnson, K D and Soukup, A A and Liu, P and Zhang, Q and Jensfold, Z and Lewis, P W and Beagrie, R A and Low, J K K and Yang, L and Mackay, J P and Godley, J A and Brand, M and Xu, J and Keles, S and Bresnick, E H},\n journal = {Journal of Clinical Investigation},\n number = {7}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"loo-waller-moreno-cole-ospinastella-pop-jochum-ali-etal-fibroblastexpressedlrrc15isareceptorforsarscov2spikeandcontrolsantiviralandantifibrotictranscriptionalprograms-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Fibroblast-expressed LRRC15 is a receptor for SARS-CoV-2 spike and controls antiviral and antifibrotic transcriptional programs.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Loo,  L.; Waller,  M.; Moreno,  C.; Cole,  A.; Ospina Stella,  A.; Pop,  O.; Jochum,  A.; Ali,  O., H.; Denes,  C.; Hamoudi,  Z.; Chung,  F.; Aggarwal,  A.; Low,  J., K., K.; Patel,  K.; Siddiquee,  R.; Kang,  T.; Mathivanan,  S.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Jochum,  W.; Flatz,  L.; Hesselson,  D.; Turville,  S., G.;  and Neely,  G., G.\n</span>\n\n  \n\n</span>\n\n  <i>PLoS Biology</i>, 21(2): e3001967.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/loo-waller-moreno-cole-ospinastella-pop-jochum-ali-etal-fibroblastexpressedlrrc15isareceptorforsarscov2spikeandcontrolsantiviralandantifibrotictranscriptionalprograms-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('loo-waller-moreno-cole-ospinastella-pop-jochum-ali-etal-fibroblastexpressedlrrc15isareceptorforsarscov2spikeandcontrolsantiviralandantifibrotictranscriptionalprograms-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Fibroblast-expressed LRRC15 is a receptor for SARS-CoV-2 spike and controls antiviral and antifibrotic transcriptional programs},\n type = {article},\n year = {2023},\n pages = {e3001967},\n volume = {21},\n id = {963b0ec7-22a6-3509-8b48-23463178bccb},\n created = {2025-05-31T09:41:42.920Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:42.920Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Loo, L and Waller, M and Moreno, C and Cole, A and Ospina Stella, A and Pop, O-T and Jochum, A-K and Ali, O H and Denes, C and Hamoudi, Z and Chung, F and Aggarwal, A and Low, J K K and Patel, K and Siddiquee, R and Kang, T and Mathivanan, S and Mackay, J P and Jochum, W and Flatz, L and Hesselson, D and Turville, S G and Neely, G G},\n journal = {PLoS Biology},\n number = {2}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"low-patel-jones-solomon-norman-maxwell-pachl-matthews-etal-mrnadisplayrevealsaclassofhighaffinitybromodomainbindingmotifsthatarenotfoundinthehumanproteome-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  mRNA display reveals a class of high-affinity bromodomain-binding motifs that are not found in the human proteome.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Low,  J., K., K.; Patel,  K.; Jones,  N.; Solomon,  P.; Norman,  A.; Maxwell,  J., W., C.; Pachl,  P.; Matthews,  J., M.; Payne,  R., J.; Passioura,  T.; Suga,  H.; Walport,  L., J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 299(12): 105482-105495.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/low-patel-jones-solomon-norman-maxwell-pachl-matthews-etal-mrnadisplayrevealsaclassofhighaffinitybromodomainbindingmotifsthatarenotfoundinthehumanproteome-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('low-patel-jones-solomon-norman-maxwell-pachl-matthews-etal-mrnadisplayrevealsaclassofhighaffinitybromodomainbindingmotifsthatarenotfoundinthehumanproteome-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {mRNA display reveals a class of high-affinity bromodomain-binding motifs that are not found in the human proteome},\n type = {article},\n year = {2023},\n pages = {105482-105495},\n volume = {299},\n id = {8138462d-5db6-3c2b-b928-0fc2e79158ae},\n created = {2025-05-31T09:41:44.027Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:44.027Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Low, J K K and Patel, K and Jones, N and Solomon, P and Norman, A and Maxwell, J W C and Pachl, P and Matthews, J M and Payne, R J and Passioura, T and Suga, H and Walport, L J and Mackay, J P},\n journal = {Journal of Biological Chemistry},\n number = {12}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"reid-low-mackay-anurdforallseasons-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A NuRD for all seasons.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Reid,  X., J.; Low,  J., K., K.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Trends Biochem Sci</i>, 48(1): 11-25.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.tibs.2022.06.002\"\n     onclick=\"log_download('reid-low-mackay-anurdforallseasons-2023', 'http://doi.org/10.1016/j.tibs.2022.06.002', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/reid-low-mackay-anurdforallseasons-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>11 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('reid-low-mackay-anurdforallseasons-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A NuRD for all seasons},\n type = {article},\n year = {2023},\n pages = {11-25},\n volume = {48},\n id = {4bc6fc94-f576-3a43-8c67-cc27c02c71e4},\n created = {2025-05-31T09:41:45.132Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:45.132Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Reid, X J and Low, J K K and Mackay, J P},\n doi = {10.1016/j.tibs.2022.06.002},\n journal = {Trends Biochem Sci},\n number = {1}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"werren-guxholli-jones-wagner-hannibal-granadillo-tyndall-moccia-etal-denovovariantsingatad2ainindividualswithdevelopmentaldisorders-2023\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  De novo variants in GATAD2A in individuals with developmental disorders.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Werren,  E., A.; Guxholli,  A.; Jones,  N.; Wagner,  M.; Hannibal,  I.; Granadillo,  J., L.; Tyndall,  A., V.; Moccia,  A.; Kuehl,  R.; Levandoski,  K., M.; Day-Salvatore,  D., L.; Wheeler,  M.; Genomics,  U., o., W., C., f., M.; Chong,  J., X.; Bamshad,  M., J.; Innes,  M.; Pierson,  T., M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Bielas,  S., L.;  and Martin,  D., M.\n</span>\n\n  \n\n</span>\n\n  <i>Human Genet Genom Adv</i>, 4(3): 100198.  2023.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/werren-guxholli-jones-wagner-hannibal-granadillo-tyndall-moccia-etal-denovovariantsingatad2ainindividualswithdevelopmentaldisorders-2023\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('werren-guxholli-jones-wagner-hannibal-granadillo-tyndall-moccia-etal-denovovariantsingatad2ainindividualswithdevelopmentaldisorders-2023')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {De novo variants in GATAD2A in individuals with developmental disorders},\n type = {article},\n year = {2023},\n pages = {100198},\n volume = {4},\n id = {32a23d36-31d5-311d-9a62-efb778e063ed},\n created = {2025-05-31T09:41:46.229Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2025-05-31T09:41:46.229Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n source_type = {Journal Article},\n private_publication = {false},\n bibtype = {article},\n author = {Werren, E A and Guxholli, A and Jones, N and Wagner, M and Hannibal, I and Granadillo, J L and Tyndall, A V and Moccia, A and Kuehl, R and Levandoski, K M and Day-Salvatore, D L and Wheeler, M and Genomics, University of Washington Center for Mendelian and Chong, J X and Bamshad, M J and Innes, M and Pierson, T M and Mackay, J P and Bielas, S L and Martin, D M},\n journal = {Human Genet Genom Adv},\n number = {3}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2022\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2022')\"\n      onkeypress=\"toggleGroup('group_2022')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2022</span>\n      <span class=\"bibbase_group_count\">\n        \n        (8)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"dowman-kulkarni-alegrerequena-giltrap-norman-sharma-gallegos-mackay-etal-siteselectivephotocatalyticfunctionalizationofpeptidesandproteinsatselenocysteine-2022\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Site-selective photocatalytic functionalization of peptides and proteins at selenocysteine.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dowman,  L.; Kulkarni,  S.; Alegre-Requena,  J.; Giltrap,  A.; Norman,  A.; Sharma,  A.; Gallegos,  L.; Mackay,  A.; Welegedara,  A.; Watson,  E.; Paton,  R.;  and Payne,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Communications</i>, 13(1).  2022.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41467-022-34530-z\"\n     onclick=\"log_download('dowman-kulkarni-alegrerequena-giltrap-norman-sharma-gallegos-mackay-etal-siteselectivephotocatalyticfunctionalizationofpeptidesandproteinsatselenocysteine-2022', 'http://doi.org/10.1038/s41467-022-34530-z', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dowman-kulkarni-alegrerequena-giltrap-norman-sharma-gallegos-mackay-etal-siteselectivephotocatalyticfunctionalizationofpeptidesandproteinsatselenocysteine-2022\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dowman-kulkarni-alegrerequena-giltrap-norman-sharma-gallegos-mackay-etal-siteselectivephotocatalyticfunctionalizationofpeptidesandproteinsatselenocysteine-2022')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Site-selective photocatalytic functionalization of peptides and proteins at selenocysteine},\n type = {article},\n year = {2022},\n volume = {13},\n id = {659f1a34-764d-3452-8843-3347bb2c9170},\n created = {2023-01-10T01:43:41.173Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:41.173Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The importance of modified peptides and proteins for applications in drug discovery, and for illuminating biological processes at the molecular level, is fueling a demand for efficient methods that facilitate the precise modification of these biomolecules. Herein, we describe the development of a photocatalytic method for the rapid and efficient dimerization and site-specific functionalization of peptide and protein diselenides. This methodology, dubbed the photocatalytic diselenide contraction, involves irradiation at 450 nm in the presence of an iridium photocatalyst and a phosphine and results in rapid and clean conversion of diselenides to reductively stable selenoethers. A mechanism for this photocatalytic transformation is proposed, which is supported by photoluminescence spectroscopy and density functional theory calculations. The utility of the photocatalytic diselenide contraction transformation is highlighted through the dimerization of selenopeptides, and by the generation of two families of protein conjugates via the site-selective modification of calmodulin containing the 21st amino acid selenocysteine, and the C-terminal modification of a ubiquitin diselenide.},\n bibtype = {article},\n author = {Dowman, L.J. and Kulkarni, S.S. and Alegre-Requena, J.V. and Giltrap, A.M. and Norman, A.R. and Sharma, A. and Gallegos, L.C. and Mackay, A.S. and Welegedara, A.P. and Watson, E.E. and Paton, R.S. and Payne, R.J.},\n doi = {10.1038/s41467-022-34530-z},\n journal = {Nature Communications},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The importance of modified peptides and proteins for applications in drug discovery, and for illuminating biological processes at the molecular level, is fueling a demand for efficient methods that facilitate the precise modification of these biomolecules. Herein, we describe the development of a photocatalytic method for the rapid and efficient dimerization and site-specific functionalization of peptide and protein diselenides. This methodology, dubbed the photocatalytic diselenide contraction, involves irradiation at 450 nm in the presence of an iridium photocatalyst and a phosphine and results in rapid and clean conversion of diselenides to reductively stable selenoethers. A mechanism for this photocatalytic transformation is proposed, which is supported by photoluminescence spectroscopy and density functional theory calculations. The utility of the photocatalytic diselenide contraction transformation is highlighted through the dimerization of selenopeptides, and by the generation of two families of protein conjugates via the site-selective modification of calmodulin containing the 21st amino acid selenocysteine, and the C-terminal modification of a ubiquitin diselenide.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"zhong-moghaddassani-paudel-low-silva-mueller-deshpande-panjikar-etal-theroleofauxiliarydomainsinmodulatingchd4activitysuggestsmechanisticcommonalitybetweenenzymefamilies-2022\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The role of auxiliary domains in modulating CHD4 activity suggests mechanistic commonality between enzyme families.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Zhong,  Y.; Moghaddas Sani,  H.; Paudel,  B.; Low,  J.; Silva,  A.; Mueller,  S.; Deshpande,  C.; Panjikar,  S.; Reid,  X.; Bedward,  M.; van Oijen,  A.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Nature Communications</i>, 13(1).  2022.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41467-022-35002-0\"\n     onclick=\"log_download('zhong-moghaddassani-paudel-low-silva-mueller-deshpande-panjikar-etal-theroleofauxiliarydomainsinmodulatingchd4activitysuggestsmechanisticcommonalitybetweenenzymefamilies-2022', 'http://doi.org/10.1038/s41467-022-35002-0', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/zhong-moghaddassani-paudel-low-silva-mueller-deshpande-panjikar-etal-theroleofauxiliarydomainsinmodulatingchd4activitysuggestsmechanisticcommonalitybetweenenzymefamilies-2022\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>4 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('zhong-moghaddassani-paudel-low-silva-mueller-deshpande-panjikar-etal-theroleofauxiliarydomainsinmodulatingchd4activitysuggestsmechanisticcommonalitybetweenenzymefamilies-2022')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The role of auxiliary domains in modulating CHD4 activity suggests mechanistic commonality between enzyme families},\n type = {article},\n year = {2022},\n volume = {13},\n id = {76602a66-c88f-3ef1-8f5b-ae0a77f5e423},\n created = {2023-01-10T01:43:42.100Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:42.100Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {CHD4 is an essential, widely conserved ATP-dependent translocase that is also a broad tumour dependency. In common with other SF2-family chromatin remodelling enzymes, it alters chromatin accessibility by repositioning histone octamers. Besides the helicase and adjacent tandem chromodomains and PHD domains, CHD4 features 1000 residues of N- and C-terminal sequence with unknown structure and function. We demonstrate that these regions regulate CHD4 activity through different mechanisms. An N-terminal intrinsically disordered region (IDR) promotes remodelling integrity in a manner that depends on the composition but not sequence of the IDR. The C-terminal region harbours an auto-inhibitory region that contacts the helicase domain. Auto-inhibition is relieved by a previously unrecognized C-terminal SANT-SLIDE domain split by ~150 residues of disordered sequence, most likely by binding of this domain to substrate DNA. Our data shed light on CHD4 regulation and reveal strong mechanistic commonality between CHD family members, as well as with ISWI-family remodellers.},\n bibtype = {article},\n author = {Zhong, Y. and Moghaddas Sani, H. and Paudel, B.P. and Low, J.K.K. and Silva, A.P.G. and Mueller, S. and Deshpande, C. and Panjikar, S. and Reid, X.J. and Bedward, M.J. and van Oijen, A.M. and Mackay, J.P.},\n doi = {10.1038/s41467-022-35002-0},\n journal = {Nature Communications},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  CHD4 is an essential, widely conserved ATP-dependent translocase that is also a broad tumour dependency. In common with other SF2-family chromatin remodelling enzymes, it alters chromatin accessibility by repositioning histone octamers. Besides the helicase and adjacent tandem chromodomains and PHD domains, CHD4 features 1000 residues of N- and C-terminal sequence with unknown structure and function. We demonstrate that these regions regulate CHD4 activity through different mechanisms. An N-terminal intrinsically disordered region (IDR) promotes remodelling integrity in a manner that depends on the composition but not sequence of the IDR. The C-terminal region harbours an auto-inhibitory region that contacts the helicase domain. Auto-inhibition is relieved by a previously unrecognized C-terminal SANT-SLIDE domain split by ~150 residues of disordered sequence, most likely by binding of this domain to substrate DNA. Our data shed light on CHD4 regulation and reveal strong mechanistic commonality between CHD family members, as well as with ISWI-family remodellers.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"ashhurst-johansen-maxwell-stockdale-ashley-aggarwal-siddiquee-miemczyk-etal-mucosaltlr2activatingproteinbasedvaccinationinducespotentpulmonaryimmunityandprotectionagainstsarscov2inmice-2022\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Mucosal TLR2-activating protein-based vaccination induces potent pulmonary immunity and protection against SARS-CoV-2 in mice.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Ashhurst,  A.; Johansen,  M.; Maxwell,  J.; Stockdale,  S.; Ashley,  C.; Aggarwal,  A.; Siddiquee,  R.; Miemczyk,  S.; Nguyen,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Payne,  R.;  and Britton,  W.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Communications</i>, 13(1).  2022.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41467-022-34297-3\"\n     onclick=\"log_download('ashhurst-johansen-maxwell-stockdale-ashley-aggarwal-siddiquee-miemczyk-etal-mucosaltlr2activatingproteinbasedvaccinationinducespotentpulmonaryimmunityandprotectionagainstsarscov2inmice-2022', 'http://doi.org/10.1038/s41467-022-34297-3', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/ashhurst-johansen-maxwell-stockdale-ashley-aggarwal-siddiquee-miemczyk-etal-mucosaltlr2activatingproteinbasedvaccinationinducespotentpulmonaryimmunityandprotectionagainstsarscov2inmice-2022\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('ashhurst-johansen-maxwell-stockdale-ashley-aggarwal-siddiquee-miemczyk-etal-mucosaltlr2activatingproteinbasedvaccinationinducespotentpulmonaryimmunityandprotectionagainstsarscov2inmice-2022')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Mucosal TLR2-activating protein-based vaccination induces potent pulmonary immunity and protection against SARS-CoV-2 in mice},\n type = {article},\n year = {2022},\n volume = {13},\n id = {817b23fa-ab01-305f-8861-84807b7e3327},\n created = {2023-01-10T01:43:43.018Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:43.018Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Current vaccines against SARS-CoV-2 substantially reduce mortality, but protection against infection is less effective. Enhancing immunity in the respiratory tract, via mucosal vaccination, may provide protection against infection and minimise viral spread. Here, we report testing of a subunit vaccine in mice, consisting of SARS-CoV-2 Spike protein with a TLR2-stimulating adjuvant (Pam2Cys), delivered to mice parenterally or mucosally. Both routes of vaccination induce substantial neutralising antibody (nAb) titres, however, mucosal vaccination uniquely generates anti-Spike IgA, increases nAb in the serum and airways, and increases lung CD4+ T-cell responses. TLR2 is expressed by respiratory epithelia and immune cells. Using TLR2 deficient chimeric mice, we determine that TLR2 expression in either compartment facilitates early innate responses to mucosal vaccination. By contrast, TLR2 on hematopoietic cells is essential for optimal lung-localised, antigen-specific responses. In K18-hACE2 mice, vaccination provides complete protection against disease and sterilising lung immunity against SARS-CoV-2, with a short-term non-specific protective effect from mucosal Pam2Cys alone. These data support mucosal vaccination as a strategy to improve protection in the respiratory tract against SARS-CoV-2 and other respiratory viruses.},\n bibtype = {article},\n author = {Ashhurst, A.S. and Johansen, M.D. and Maxwell, J.W.C. and Stockdale, S. and Ashley, C.L. and Aggarwal, A. and Siddiquee, R. and Miemczyk, S. and Nguyen, D.H. and Mackay, J.P. and Payne, R.J. and Britton, W.J.},\n doi = {10.1038/s41467-022-34297-3},\n journal = {Nature Communications},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Current vaccines against SARS-CoV-2 substantially reduce mortality, but protection against infection is less effective. Enhancing immunity in the respiratory tract, via mucosal vaccination, may provide protection against infection and minimise viral spread. Here, we report testing of a subunit vaccine in mice, consisting of SARS-CoV-2 Spike protein with a TLR2-stimulating adjuvant (Pam2Cys), delivered to mice parenterally or mucosally. Both routes of vaccination induce substantial neutralising antibody (nAb) titres, however, mucosal vaccination uniquely generates anti-Spike IgA, increases nAb in the serum and airways, and increases lung CD4+ T-cell responses. TLR2 is expressed by respiratory epithelia and immune cells. Using TLR2 deficient chimeric mice, we determine that TLR2 expression in either compartment facilitates early innate responses to mucosal vaccination. By contrast, TLR2 on hematopoietic cells is essential for optimal lung-localised, antigen-specific responses. In K18-hACE2 mice, vaccination provides complete protection against disease and sterilising lung immunity against SARS-CoV-2, with a short-term non-specific protective effect from mucosal Pam2Cys alone. These data support mucosal vaccination as a strategy to improve protection in the respiratory tract against SARS-CoV-2 and other respiratory viruses.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"ullah-thlken-zhong-john-rossbach-lenz-gringer-nist-etal-rnainhibitsdmi2chd4chromatinbindingandnucleosomeremodeling-2022\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  RNA inhibits dMi-2/CHD4 chromatin binding and nucleosome remodeling.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Ullah,  I.; Thölken,  C.; Zhong,  Y.; John,  M.; Rossbach,  O.; Lenz,  J.; Gößringer,  M.; Nist,  A.; Albert,  L.; Stiewe,  T.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Brehm,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Cell Reports</i>, 39(9).  2022.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.celrep.2022.110895\"\n     onclick=\"log_download('ullah-thlken-zhong-john-rossbach-lenz-gringer-nist-etal-rnainhibitsdmi2chd4chromatinbindingandnucleosomeremodeling-2022', 'http://doi.org/10.1016/j.celrep.2022.110895', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/ullah-thlken-zhong-john-rossbach-lenz-gringer-nist-etal-rnainhibitsdmi2chd4chromatinbindingandnucleosomeremodeling-2022\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('ullah-thlken-zhong-john-rossbach-lenz-gringer-nist-etal-rnainhibitsdmi2chd4chromatinbindingandnucleosomeremodeling-2022')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {RNA inhibits dMi-2/CHD4 chromatin binding and nucleosome remodeling},\n type = {article},\n year = {2022},\n volume = {39},\n id = {d99d4ee9-901c-3e7f-9a93-d47a33ffacef},\n created = {2023-01-10T01:43:43.942Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:43.942Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The ATP-dependent nucleosome remodeler Mi-2/CHD4 broadly modulates chromatin landscapes to repress transcription and to maintain genome integrity. Here we use individual nucleotide resolution crosslinking and immunoprecipitation (iCLIP) to show that Drosophila Mi-2 associates with thousands of mRNA molecules in vivo. Biochemical data reveal that recombinant dMi-2 preferentially binds to G-rich RNA molecules using two intrinsically disordered regions of unclear function. Pharmacological inhibition of transcription and RNase digestion approaches establish that RNA inhibits the association of dMi-2 with chromatin. We also show that RNA inhibits dMi-2-mediated nucleosome mobilization by competing with the nucleosome substrate. Importantly, this activity is shared by CHD4, the human homolog of dMi-2, strongly suggesting that RNA-mediated regulation of remodeler activity is an evolutionary conserved mechanism. Our data support a model in which RNA serves to protect actively transcribed regions of the genome from dMi-2/CHD4-mediated establishment of repressive chromatin structures.},\n bibtype = {article},\n author = {Ullah, I. and Thölken, C. and Zhong, Y. and John, M. and Rossbach, O. and Lenz, J. and Gößringer, M. and Nist, A. and Albert, L. and Stiewe, T. and Mackay, J.P. and Brehm, A.},\n doi = {10.1016/j.celrep.2022.110895},\n journal = {Cell Reports},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The ATP-dependent nucleosome remodeler Mi-2/CHD4 broadly modulates chromatin landscapes to repress transcription and to maintain genome integrity. Here we use individual nucleotide resolution crosslinking and immunoprecipitation (iCLIP) to show that Drosophila Mi-2 associates with thousands of mRNA molecules in vivo. Biochemical data reveal that recombinant dMi-2 preferentially binds to G-rich RNA molecules using two intrinsically disordered regions of unclear function. Pharmacological inhibition of transcription and RNase digestion approaches establish that RNA inhibits the association of dMi-2 with chromatin. We also show that RNA inhibits dMi-2-mediated nucleosome mobilization by competing with the nucleosome substrate. Importantly, this activity is shared by CHD4, the human homolog of dMi-2, strongly suggesting that RNA-mediated regulation of remodeler activity is an evolutionary conserved mechanism. Our data support a model in which RNA serves to protect actively transcribed regions of the genome from dMi-2/CHD4-mediated establishment of repressive chromatin structures.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"hodson-low-vantwest-jones-swuec-murphy-tsukada-fawkes-etal-mechanismofbloomsyndromecomplexassemblyrequiredfordoublehollidayjunctiondissolutionandgenomestability-2022\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Mechanism of Bloom syndrome complex assembly required for double Holliday junction dissolution and genome stability.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Hodson,  C.; Low,  J.; van Twest,  S.; Jones,  S.; Swuec,  P.; Murphy,  V.; Tsukada,  K.; Fawkes,  M.; Bythell-Douglas,  R.; Davies,  A.; Costa,  A.;  and Deans,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 119(6).  2022.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.2109093119\"\n     onclick=\"log_download('hodson-low-vantwest-jones-swuec-murphy-tsukada-fawkes-etal-mechanismofbloomsyndromecomplexassemblyrequiredfordoublehollidayjunctiondissolutionandgenomestability-2022', 'http://doi.org/10.1073/pnas.2109093119', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/hodson-low-vantwest-jones-swuec-murphy-tsukada-fawkes-etal-mechanismofbloomsyndromecomplexassemblyrequiredfordoublehollidayjunctiondissolutionandgenomestability-2022\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('hodson-low-vantwest-jones-swuec-murphy-tsukada-fawkes-etal-mechanismofbloomsyndromecomplexassemblyrequiredfordoublehollidayjunctiondissolutionandgenomestability-2022')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Mechanism of Bloom syndrome complex assembly required for double Holliday junction dissolution and genome stability},\n type = {article},\n year = {2022},\n volume = {119},\n id = {eea6affc-fd8d-3936-9453-941dacf51a3d},\n created = {2023-01-10T01:43:44.899Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:44.899Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The RecQ-like helicase BLM cooperates with topoisomerase IIIα, RMI1, and RMI2 in a heterotetrameric complex (the “Bloom syndrome complex”) for dissolution of double Holliday junctions, key intermediates in homologous recombination. Mutations in any component of the Bloom syndrome complex can cause genome instability and a highly cancer-prone disorder called Bloom syndrome. Some heterozygous carriers are also predisposed to breast cancer. To understand how the activities of BLM helicase and topoisomerase IIIα are coupled, we purified the active four-subunit complex. Chemical cross-linking and mass spectrometry revealed a unique architecture that links the helicase and topoisomerase domains. Using biochemical experiments, we demonstrated dimerization mediated by the N terminus of BLM with a 2:2:2:2 stoichiometry within the Bloom syndrome complex. We identified mutations that independently abrogate dimerization or association of BLM with RMI1, and we show that both are dysfunctional for dissolution using in vitro assays and cause genome instability and synthetic lethal interactions with GEN1/MUS81 in cells. Truncated BLM can also inhibit the activity of full-length BLM in mixed dimers, suggesting a putative mechanism of dominant-negative action in carriers of BLM truncation alleles. Our results identify critical molecular determinants of Bloom syndrome complex assembly required for double Holliday junction dissolution and maintenance of genome stability.},\n bibtype = {article},\n author = {Hodson, C. and Low, J.K.K. and van Twest, S. and Jones, S.E. and Swuec, P. and Murphy, V. and Tsukada, K. and Fawkes, M. and Bythell-Douglas, R. and Davies, A. and Costa, A. and Deans, A.J.},\n doi = {10.1073/pnas.2109093119},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The RecQ-like helicase BLM cooperates with topoisomerase IIIα, RMI1, and RMI2 in a heterotetrameric complex (the “Bloom syndrome complex”) for dissolution of double Holliday junctions, key intermediates in homologous recombination. Mutations in any component of the Bloom syndrome complex can cause genome instability and a highly cancer-prone disorder called Bloom syndrome. Some heterozygous carriers are also predisposed to breast cancer. To understand how the activities of BLM helicase and topoisomerase IIIα are coupled, we purified the active four-subunit complex. Chemical cross-linking and mass spectrometry revealed a unique architecture that links the helicase and topoisomerase domains. Using biochemical experiments, we demonstrated dimerization mediated by the N terminus of BLM with a 2:2:2:2 stoichiometry within the Bloom syndrome complex. We identified mutations that independently abrogate dimerization or association of BLM with RMI1, and we show that both are dysfunctional for dissolution using in vitro assays and cause genome instability and synthetic lethal interactions with GEN1/MUS81 in cells. Truncated BLM can also inhibit the activity of full-length BLM in mixed dimers, suggesting a putative mechanism of dominant-negative action in carriers of BLM truncation alleles. Our results identify critical molecular determinants of Bloom syndrome complex assembly required for double Holliday junction dissolution and maintenance of genome stability.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"arvindekar-jackman-low-landsberg-mackay-viswanath-moleculararchitectureofnucleosomeremodelinganddeacetylasesubcomplexesbyintegrativestructuredetermination-2022\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Molecular architecture of nucleosome remodeling and deacetylase sub-complexes by integrative structure determination.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Arvindekar,  S.; Jackman,  M.; Low,  J.; Landsberg,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Viswanath,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 31(9).  2022.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.4387\"\n     onclick=\"log_download('arvindekar-jackman-low-landsberg-mackay-viswanath-moleculararchitectureofnucleosomeremodelinganddeacetylasesubcomplexesbyintegrativestructuredetermination-2022', 'http://doi.org/10.1002/pro.4387', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/arvindekar-jackman-low-landsberg-mackay-viswanath-moleculararchitectureofnucleosomeremodelinganddeacetylasesubcomplexesbyintegrativestructuredetermination-2022\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('arvindekar-jackman-low-landsberg-mackay-viswanath-moleculararchitectureofnucleosomeremodelinganddeacetylasesubcomplexesbyintegrativestructuredetermination-2022')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Molecular architecture of nucleosome remodeling and deacetylase sub-complexes by integrative structure determination},\n type = {article},\n year = {2022},\n volume = {31},\n id = {eef4f63c-8daa-3623-b17b-a5cf28db9aee},\n created = {2023-01-10T01:43:45.810Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:45.810Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a chromatin-modifying assembly that regulates gene expression and DNA damage repair. Despite its importance, limited structural information describing the complete NuRD complex is available and a detailed understanding of its mechanism is therefore lacking. Drawing on information from SEC-MALLS, DIA-MS, XLMS, negative-stain EM, X-ray crystallography, NMR spectroscopy, secondary structure predictions, and homology models, we applied Bayesian integrative structure determination to investigate the molecular architecture of three NuRD sub-complexes: MTA1-HDAC1-RBBP4, MTA1N-HDAC1-MBD3GATAD2CC, and MTA1-HDAC1-RBBP4-MBD3-GATAD2A [nucleosome deacetylase (NuDe)]. The integrative structures were corroborated by examining independent crosslinks, cryo-EM maps, biochemical assays, known cancer-associated mutations, and structure predictions from AlphaFold. The robustness of the models was assessed by jack-knifing. Localization of the full-length MBD3, which connects the deacetylase and chromatin remodeling modules in NuRD, has not previously been possible; our models indicate two different locations for MBD3, suggesting a mechanism by which MBD3 in the presence of GATAD2A asymmetrically bridges the two modules in NuRD. Further, our models uncovered three previously unrecognized subunit interfaces in NuDe: HDAC1C-MTA1BAH, MTA1BAH-MBD3MBD, and HDAC160–100-MBD3MBD. Our approach also allowed us to localize regions of unknown structure, such as HDAC1C and MBD3IDR, thereby resulting in the most complete and robustly cross-validated structural characterization of these NuRD sub-complexes so far.},\n bibtype = {article},\n author = {Arvindekar, S. and Jackman, M.J. and Low, J.K.K. and Landsberg, M.J. and Mackay, J.P. and Viswanath, S.},\n doi = {10.1002/pro.4387},\n journal = {Protein Science},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nucleosome remodeling and deacetylase (NuRD) complex is a chromatin-modifying assembly that regulates gene expression and DNA damage repair. Despite its importance, limited structural information describing the complete NuRD complex is available and a detailed understanding of its mechanism is therefore lacking. Drawing on information from SEC-MALLS, DIA-MS, XLMS, negative-stain EM, X-ray crystallography, NMR spectroscopy, secondary structure predictions, and homology models, we applied Bayesian integrative structure determination to investigate the molecular architecture of three NuRD sub-complexes: MTA1-HDAC1-RBBP4, MTA1N-HDAC1-MBD3GATAD2CC, and MTA1-HDAC1-RBBP4-MBD3-GATAD2A [nucleosome deacetylase (NuDe)]. The integrative structures were corroborated by examining independent crosslinks, cryo-EM maps, biochemical assays, known cancer-associated mutations, and structure predictions from AlphaFold. The robustness of the models was assessed by jack-knifing. Localization of the full-length MBD3, which connects the deacetylase and chromatin remodeling modules in NuRD, has not previously been possible; our models indicate two different locations for MBD3, suggesting a mechanism by which MBD3 in the presence of GATAD2A asymmetrically bridges the two modules in NuRD. Further, our models uncovered three previously unrecognized subunit interfaces in NuDe: HDAC1C-MTA1BAH, MTA1BAH-MBD3MBD, and HDAC160–100-MBD3MBD. Our approach also allowed us to localize regions of unknown structure, such as HDAC1C and MBD3IDR, thereby resulting in the most complete and robustly cross-validated structural characterization of these NuRD sub-complexes so far.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"desmarini-truong-wilkinsonwhite-desphande-torrado-mackay-matthews-sorrell-etal-tnpanaloguesinhibitthevirulencepromotingipinf34infkinasearg1inthefungalpathogencryptococcusneoformans-2022\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  TNP Analogues Inhibit the Virulence Promoting IP<inf>3-4</inf> Kinase Arg1 in the Fungal Pathogen Cryptococcus neoformans.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Desmarini,  D.; Truong,  D.; Wilkinson-White,  L.; Desphande,  C.; Torrado,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Matthews,  J.; Sorrell,  T.; Lev,  S.; Thompson,  P.; Thompson,  P.;  and Djordjevic,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecules</i>, 12(10).  2022.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.3390/biom12101526\"\n     onclick=\"log_download('desmarini-truong-wilkinsonwhite-desphande-torrado-mackay-matthews-sorrell-etal-tnpanaloguesinhibitthevirulencepromotingipinf34infkinasearg1inthefungalpathogencryptococcusneoformans-2022', 'http://doi.org/10.3390/biom12101526', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/desmarini-truong-wilkinsonwhite-desphande-torrado-mackay-matthews-sorrell-etal-tnpanaloguesinhibitthevirulencepromotingipinf34infkinasearg1inthefungalpathogencryptococcusneoformans-2022\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('desmarini-truong-wilkinsonwhite-desphande-torrado-mackay-matthews-sorrell-etal-tnpanaloguesinhibitthevirulencepromotingipinf34infkinasearg1inthefungalpathogencryptococcusneoformans-2022')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {TNP Analogues Inhibit the Virulence Promoting IP&lt;inf&gt;3-4&lt;/inf&gt; Kinase Arg1 in the Fungal Pathogen Cryptococcus neoformans},\n type = {article},\n year = {2022},\n volume = {12},\n id = {9cbc3fce-c9c1-35f8-ba6d-09bac2420d29},\n created = {2023-01-10T01:43:46.835Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:46.835Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {New antifungals with unique modes of action are urgently needed to treat the increasing global burden of invasive fungal infections. The fungal inositol polyphosphate kinase (IPK) pathway, comprised of IPKs that convert IP3 to IP8, provides a promising new target due to its impact on multiple, critical cellular functions and, unlike in mammalian cells, its lack of redundancy. Nearly all IPKs in the fungal pathway are essential for virulence, with IP3-4 kinase (IP3-4K) the most critical. The dibenzylaminopurine compound, N2-(m-trifluorobenzylamino)-N6-(p-nitrobenzylamino)purine (TNP), is a commercially available inhibitor of mammalian IPKs. The ability of TNP to be adapted as an inhibitor of fungal IP3-4K has not been investigated. We purified IP3-4K from the human pathogens, Cryptococcus neoformans and Candida albicans, and optimised enzyme and surface plasmon resonance (SPR) assays to determine the half inhibitory concentration (IC50) and binding affinity (KD), respectively, of TNP and 38 analogues. A novel chemical route was developed to efficiently prepare TNP analogues. TNP and its analogues demonstrated inhibition of recombinant IP3-4K from C. neoformans (CnArg1) at low µM IC50s, but not IP3-4K from C. albicans (CaIpk2) and many analogues exhibited selectivity for CnArg1 over the human equivalent, HsIPMK. Our results provide a foundation for improving potency and selectivity of the TNP series for fungal IP3-4K.},\n bibtype = {article},\n author = {Desmarini, D. and Truong, D. and Wilkinson-White, L. and Desphande, C. and Torrado, M. and Mackay, J.P. and Matthews, J.M. and Sorrell, T.C. and Lev, S. and Thompson, P.E. and Thompson, P.E. and Djordjevic, J.T.},\n doi = {10.3390/biom12101526},\n journal = {Biomolecules},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  New antifungals with unique modes of action are urgently needed to treat the increasing global burden of invasive fungal infections. The fungal inositol polyphosphate kinase (IPK) pathway, comprised of IPKs that convert IP3 to IP8, provides a promising new target due to its impact on multiple, critical cellular functions and, unlike in mammalian cells, its lack of redundancy. Nearly all IPKs in the fungal pathway are essential for virulence, with IP3-4 kinase (IP3-4K) the most critical. The dibenzylaminopurine compound, N2-(m-trifluorobenzylamino)-N6-(p-nitrobenzylamino)purine (TNP), is a commercially available inhibitor of mammalian IPKs. The ability of TNP to be adapted as an inhibitor of fungal IP3-4K has not been investigated. We purified IP3-4K from the human pathogens, Cryptococcus neoformans and Candida albicans, and optimised enzyme and surface plasmon resonance (SPR) assays to determine the half inhibitory concentration (IC50) and binding affinity (KD), respectively, of TNP and 38 analogues. A novel chemical route was developed to efficiently prepare TNP analogues. TNP and its analogues demonstrated inhibition of recombinant IP3-4K from C. neoformans (CnArg1) at low µM IC50s, but not IP3-4K from C. albicans (CaIpk2) and many analogues exhibited selectivity for CnArg1 over the human equivalent, HsIPMK. Our results provide a foundation for improving potency and selectivity of the TNP series for fungal IP3-4K.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"sharifitabar-giardina-feng-francis-moghaddassani-low-mackay-bailey-etal-uniqueproteininteractionnetworksdefinethechromatinremodellingmoduleofthenurdcomplex-2022\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Unique protein interaction networks define the chromatin remodelling module of the NuRD complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Sharifi Tabar,  M.; Giardina,  C.; Feng,  Y.; Francis,  H.; Moghaddas Sani,  H.; Low,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Bailey,  C.;  and Rasko,  J.\n</span>\n\n  \n\n</span>\n\n  <i>FEBS Journal</i>, 289(1): 199-214.  2022.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1111/febs.16112\"\n     onclick=\"log_download('sharifitabar-giardina-feng-francis-moghaddassani-low-mackay-bailey-etal-uniqueproteininteractionnetworksdefinethechromatinremodellingmoduleofthenurdcomplex-2022', 'http://doi.org/10.1111/febs.16112', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/sharifitabar-giardina-feng-francis-moghaddassani-low-mackay-bailey-etal-uniqueproteininteractionnetworksdefinethechromatinremodellingmoduleofthenurdcomplex-2022\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('sharifitabar-giardina-feng-francis-moghaddassani-low-mackay-bailey-etal-uniqueproteininteractionnetworksdefinethechromatinremodellingmoduleofthenurdcomplex-2022')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Unique protein interaction networks define the chromatin remodelling module of the NuRD complex},\n type = {article},\n year = {2022},\n pages = {199-214},\n volume = {289},\n id = {a6268275-0bc8-3002-bce9-6d7a08deb3b1},\n created = {2023-01-10T01:43:47.765Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:47.765Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The combination of four proteins and their paralogues including MBD2/3, GATAD2A/B, CDK2AP1 and CHD3/4/5, which we refer to as the MGCC module, form the chromatin remodelling module of the nucleosome remodelling and deacetylase (NuRD) complex. To date, mechanisms by which the MGCC module acquires paralogue-specific function and specificity have not been addressed. Understanding the protein–protein interaction (PPI) network of the MGCC subunits is essential for defining underlying mechanisms of gene regulation. Therefore, using pulldown followed by mass spectrometry analysis (PD-MS), we report a proteome-wide interaction network of the MGCC module in a paralogue-specific manner. Our data also demonstrate that the disordered C-terminal region of CHD3/4/5 is a gateway to incorporate remodelling activity into both ChAHP (CHD4, ADNP, HP1γ) and NuRD complexes in a mutually exclusive manner. We define a short aggregation-prone region (APR) within the C-terminal segment of GATAD2B that is essential for the interaction of CHD4 and CDK2AP1 with the NuRD complex. Finally, we also report an association of CDK2AP1 with the nuclear receptor co-repressor (NCOR) complex. Overall, this study provides insight into the possible mechanisms through which the MGCC module can achieve specificity and diverse biological functions.},\n bibtype = {article},\n author = {Sharifi Tabar, M. and Giardina, C. and Feng, Y. and Francis, H. and Moghaddas Sani, H. and Low, J.K.K. and Mackay, J.P. and Bailey, C.G. and Rasko, J.E.J.},\n doi = {10.1111/febs.16112},\n journal = {FEBS Journal},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The combination of four proteins and their paralogues including MBD2/3, GATAD2A/B, CDK2AP1 and CHD3/4/5, which we refer to as the MGCC module, form the chromatin remodelling module of the nucleosome remodelling and deacetylase (NuRD) complex. To date, mechanisms by which the MGCC module acquires paralogue-specific function and specificity have not been addressed. Understanding the protein–protein interaction (PPI) network of the MGCC subunits is essential for defining underlying mechanisms of gene regulation. Therefore, using pulldown followed by mass spectrometry analysis (PD-MS), we report a proteome-wide interaction network of the MGCC module in a paralogue-specific manner. Our data also demonstrate that the disordered C-terminal region of CHD3/4/5 is a gateway to incorporate remodelling activity into both ChAHP (CHD4, ADNP, HP1γ) and NuRD complexes in a mutually exclusive manner. We define a short aggregation-prone region (APR) within the C-terminal segment of GATAD2B that is essential for the interaction of CHD4 and CDK2AP1 with the NuRD complex. Finally, we also report an association of CDK2AP1 with the nuclear receptor co-repressor (NCOR) complex. Overall, this study provides insight into the possible mechanisms through which the MGCC module can achieve specificity and diverse biological functions.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2021\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2021')\"\n      onkeypress=\"toggleGroup('group_2021')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2021</span>\n      <span class=\"bibbase_group_count\">\n        \n        (7)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"patel-solomon-walshe-ford-wilkinsonwhite-payne-low-mackay-betfamilybromodomainscanrecognizediacetylatedsequencesfromtranscriptionfactorsusingaconservedmechanism-2021\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  BET-Family Bromodomains Can Recognize Diacetylated Sequences from Transcription Factors Using a Conserved Mechanism.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Patel,  K.; Solomon,  P.; Walshe,  J.; Ford,  D.; Wilkinson-White,  L.; Payne,  R.; Low,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 60(9): 648-662.  2021.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/acs.biochem.0c00816\"\n     onclick=\"log_download('patel-solomon-walshe-ford-wilkinsonwhite-payne-low-mackay-betfamilybromodomainscanrecognizediacetylatedsequencesfromtranscriptionfactorsusingaconservedmechanism-2021', 'http://doi.org/10.1021/acs.biochem.0c00816', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/patel-solomon-walshe-ford-wilkinsonwhite-payne-low-mackay-betfamilybromodomainscanrecognizediacetylatedsequencesfromtranscriptionfactorsusingaconservedmechanism-2021\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('patel-solomon-walshe-ford-wilkinsonwhite-payne-low-mackay-betfamilybromodomainscanrecognizediacetylatedsequencesfromtranscriptionfactorsusingaconservedmechanism-2021')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {BET-Family Bromodomains Can Recognize Diacetylated Sequences from Transcription Factors Using a Conserved Mechanism},\n type = {article},\n year = {2021},\n pages = {648-662},\n volume = {60},\n id = {50cd9f3e-3d6d-35ea-b47c-1090ed52dff5},\n created = {2023-01-10T01:43:48.713Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:48.713Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Almost all eukaryotic proteins receive diverse post-translational modifications (PTMs) that modulate protein activity. Many histone PTMs are well characterized, heavily influence gene regulation, and are often predictors of distinct transcriptional programs. Although our understanding of the histone PTM network has matured, much is yet to be understood about the roles of transcription factor (TF) PTMs, which might well represent a similarly complex and dynamic network of functional regulation. Members of the bromodomain and extra-terminal domain (BET) family of proteins recognize acetyllysine residues and relay the signals encoded by these modifications. Here, we have investigated the acetylation dependence of several functionally relevant BET-TF interactions in vitro using surface plasmon resonance, nuclear magnetic resonance, and X-ray crystallography. We show that motifs known to be acetylated in TFs E2F1 and MyoD1 can interact with all bromodomains of BRD2, BRD3, and BRD4. The interactions are dependent on diacetylation of the motifs and show a preference for the first BET bromodomain. Structural mapping of the interactions confirms a conserved mode of binding for the two TFs to the acetyllysine binding pocket of the BET bromodomains, mimicking that of other already established functionally important histone- and TF-BET interactions. We also examined a motif from the TF RelA that is known to be acetylated but were unable to observe any interaction, regardless of the acetylation state of the sequence. Our findings overall advance our understanding of BET-TF interactions and suggest a physical link between the important diacetylated motifs found in E2F1 and MyoD1 and the BET-family proteins.},\n bibtype = {article},\n author = {Patel, K. and Solomon, P.D. and Walshe, J.L. and Ford, D.J. and Wilkinson-White, L. and Payne, R.J. and Low, J.K.K. and Mackay, J.P.},\n doi = {10.1021/acs.biochem.0c00816},\n journal = {Biochemistry},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Almost all eukaryotic proteins receive diverse post-translational modifications (PTMs) that modulate protein activity. Many histone PTMs are well characterized, heavily influence gene regulation, and are often predictors of distinct transcriptional programs. Although our understanding of the histone PTM network has matured, much is yet to be understood about the roles of transcription factor (TF) PTMs, which might well represent a similarly complex and dynamic network of functional regulation. Members of the bromodomain and extra-terminal domain (BET) family of proteins recognize acetyllysine residues and relay the signals encoded by these modifications. Here, we have investigated the acetylation dependence of several functionally relevant BET-TF interactions in vitro using surface plasmon resonance, nuclear magnetic resonance, and X-ray crystallography. We show that motifs known to be acetylated in TFs E2F1 and MyoD1 can interact with all bromodomains of BRD2, BRD3, and BRD4. The interactions are dependent on diacetylation of the motifs and show a preference for the first BET bromodomain. Structural mapping of the interactions confirms a conserved mode of binding for the two TFs to the acetyllysine binding pocket of the BET bromodomains, mimicking that of other already established functionally important histone- and TF-BET interactions. We also examined a motif from the TF RelA that is known to be acetylated but were unable to observe any interaction, regardless of the acetylation state of the sequence. Our findings overall advance our understanding of BET-TF interactions and suggest a physical link between the important diacetylated motifs found in E2F1 and MyoD1 and the BET-family proteins.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"walport-low-matthews-mackay-thecharacterizationofproteininteractionswhathowandhowmuch-2021\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The characterization of protein interactions-what, how and how much?.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Walport,  L.; Low,  J.; Matthews,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Chemical Society Reviews</i>, 50(22): 12292-12307.  2021.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1039/d1cs00548k\"\n     onclick=\"log_download('walport-low-matthews-mackay-thecharacterizationofproteininteractionswhathowandhowmuch-2021', 'http://doi.org/10.1039/d1cs00548k', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/walport-low-matthews-mackay-thecharacterizationofproteininteractionswhathowandhowmuch-2021\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>4 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('walport-low-matthews-mackay-thecharacterizationofproteininteractionswhathowandhowmuch-2021')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The characterization of protein interactions-what, how and how much?},\n type = {article},\n year = {2021},\n pages = {12292-12307},\n volume = {50},\n id = {2d324f7d-025f-3a4a-af9f-b3ca4892be6b},\n created = {2023-01-10T01:43:49.626Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:49.626Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Protein interactions underlie most molecular events in biology. Many methods have been developed to identify protein partners, to measure the affinity with which these biomolecules interact and to characterise the structures of the complexes. Each approach has its own advantages and limitations, and it can be difficult for the newcomer to determine which methodology would best suit their system. This review provides an overview of many of the techniques most widely used to identify protein partners, assess stoichiometry and binding affinity, and determine low-resolution models for complexes. Key methods covered include: yeast two-hybrid analysis, affinity purification mass spectrometry and proximity labelling to identify partners; size-exclusion chromatography, scattering methods, native mass spectrometry and analytical ultracentrifugation to estimate stoichiometry; isothermal titration calorimetry, biosensors and fluorometric methods (including microscale thermophoresis, anisotropy/polarisation, resonance energy transfer, AlphaScreen, and differential scanning fluorimetry) to measure binding affinity; and crosslinking and hydrogen-deuterium exchange mass spectrometry to probe the structure of complexes. This journal is},\n bibtype = {article},\n author = {Walport, L.J. and Low, J.K.K. and Matthews, J.M. and MacKay, J.P.},\n doi = {10.1039/d1cs00548k},\n journal = {Chemical Society Reviews},\n number = {22}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Protein interactions underlie most molecular events in biology. Many methods have been developed to identify protein partners, to measure the affinity with which these biomolecules interact and to characterise the structures of the complexes. Each approach has its own advantages and limitations, and it can be difficult for the newcomer to determine which methodology would best suit their system. This review provides an overview of many of the techniques most widely used to identify protein partners, assess stoichiometry and binding affinity, and determine low-resolution models for complexes. Key methods covered include: yeast two-hybrid analysis, affinity purification mass spectrometry and proximity labelling to identify partners; size-exclusion chromatography, scattering methods, native mass spectrometry and analytical ultracentrifugation to estimate stoichiometry; isothermal titration calorimetry, biosensors and fluorometric methods (including microscale thermophoresis, anisotropy/polarisation, resonance energy transfer, AlphaScreen, and differential scanning fluorimetry) to measure binding affinity; and crosslinking and hydrogen-deuterium exchange mass spectrometry to probe the structure of complexes. This journal is\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"patel-solomon-walshe-low-mackay-thebromodomainsofbetfamilyproteinscanrecognizediacetylatedhistoneh2az-2021\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The bromodomains of BET family proteins can recognize diacetylated histone H2A.Z.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Patel,  K.; Solomon,  P.; Walshe,  J.; Low,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 30(2): 464-476.  2021.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.4006\"\n     onclick=\"log_download('patel-solomon-walshe-low-mackay-thebromodomainsofbetfamilyproteinscanrecognizediacetylatedhistoneh2az-2021', 'http://doi.org/10.1002/pro.4006', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/patel-solomon-walshe-low-mackay-thebromodomainsofbetfamilyproteinscanrecognizediacetylatedhistoneh2az-2021\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('patel-solomon-walshe-low-mackay-thebromodomainsofbetfamilyproteinscanrecognizediacetylatedhistoneh2az-2021')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The bromodomains of BET family proteins can recognize diacetylated histone H2A.Z},\n type = {article},\n year = {2021},\n pages = {464-476},\n volume = {30},\n id = {b502f967-797b-3fb0-8bb3-8923a5fa0bbd},\n created = {2023-01-10T01:43:50.588Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:50.588Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Chemical modifications of histone tails influence genome accessibility and the transcriptional state of eukaryotic cells. Lysine acetylation is one of the most common modifications and acetyllysine-binding bromodomains (BDs) provide a means for acetyllysine marks to be translated into meaningful cellular responses. Here, we have investigated the mechanism underlying the reported association between the Bromodomain and Extra Terminal (BET) family of BD proteins and the essential histone variant H2A.Z. We use NMR spectroscopy to demonstrate a physical interaction between the N-terminal tail of H2A.Z and the BDs of BRD2, BRD3, and BRD4, and show that the interaction is dependent on lysine acetylation in H2A.Z. The BDs preferentially engage a diacetylated H2A.Z-K4acK7ac motif that is reminiscent of sequences found in other biologically important BET BD target proteins, including histones and transcription factors. A H2A.Z-K7acK11ac motif can also bind BET BDs—with a preference for the second BD of each protein. Chemical shift perturbation mapping of the interactions, together with an X-ray crystal structure of BRD2-BD1 bound to H2A.Z-K4acK7ac, shows that H2A.Z binds the canonical AcK binding pocket of the BDs. This mechanism mirrors the conserved binding mode that is unique to the BET BDs, in which two acetylation marks are read simultaneously by a single BD. Our findings provide structural corroboration of biochemical and cell biological data that link H2A.Z and BET-family proteins, suggesting that the function of H2A.Z is enacted through interactions with these chromatin readers.},\n bibtype = {article},\n author = {Patel, K. and Solomon, P.D. and Walshe, J.L. and Low, J.K.K. and Mackay, J.P.},\n doi = {10.1002/pro.4006},\n journal = {Protein Science},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chemical modifications of histone tails influence genome accessibility and the transcriptional state of eukaryotic cells. Lysine acetylation is one of the most common modifications and acetyllysine-binding bromodomains (BDs) provide a means for acetyllysine marks to be translated into meaningful cellular responses. Here, we have investigated the mechanism underlying the reported association between the Bromodomain and Extra Terminal (BET) family of BD proteins and the essential histone variant H2A.Z. We use NMR spectroscopy to demonstrate a physical interaction between the N-terminal tail of H2A.Z and the BDs of BRD2, BRD3, and BRD4, and show that the interaction is dependent on lysine acetylation in H2A.Z. The BDs preferentially engage a diacetylated H2A.Z-K4acK7ac motif that is reminiscent of sequences found in other biologically important BET BD target proteins, including histones and transcription factors. A H2A.Z-K7acK11ac motif can also bind BET BDs—with a preference for the second BD of each protein. Chemical shift perturbation mapping of the interactions, together with an X-ray crystal structure of BRD2-BD1 bound to H2A.Z-K4acK7ac, shows that H2A.Z binds the canonical AcK binding pocket of the BDs. This mechanism mirrors the conserved binding mode that is unique to the BET BDs, in which two acetylation marks are read simultaneously by a single BD. Our findings provide structural corroboration of biochemical and cell biological data that link H2A.Z and BET-family proteins, suggesting that the function of H2A.Z is enacted through interactions with these chromatin readers.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"byrne-bedding-corcilius-ford-zhong-franck-larance-mackay-etal-latestagemodificationofpeptidesandproteinsatcysteinewithdiaryliodoniumsalts-2021\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Late-stage modification of peptides and proteins at cysteine with diaryliodonium salts.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Byrne,  S.; Bedding,  M.; Corcilius,  L.; Ford,  D.; Zhong,  Y.; Franck,  C.; Larance,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Payne,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Chemical Science</i>, 12(42): 14159-14166.  2021.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1039/d1sc03127a\"\n     onclick=\"log_download('byrne-bedding-corcilius-ford-zhong-franck-larance-mackay-etal-latestagemodificationofpeptidesandproteinsatcysteinewithdiaryliodoniumsalts-2021', 'http://doi.org/10.1039/d1sc03127a', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/byrne-bedding-corcilius-ford-zhong-franck-larance-mackay-etal-latestagemodificationofpeptidesandproteinsatcysteinewithdiaryliodoniumsalts-2021\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('byrne-bedding-corcilius-ford-zhong-franck-larance-mackay-etal-latestagemodificationofpeptidesandproteinsatcysteinewithdiaryliodoniumsalts-2021')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Late-stage modification of peptides and proteins at cysteine with diaryliodonium salts},\n type = {article},\n year = {2021},\n pages = {14159-14166},\n volume = {12},\n id = {fe1847bf-a481-3fe2-9167-b70bf15b8ea0},\n created = {2023-01-10T01:43:51.522Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:51.522Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The modification of peptides and proteins has emerged as a powerful means to efficiently prepare high value bioconjugates for a range of applications in chemical biology and for the development of next-generation therapeutics. Herein, we report a novel method for the chemoselective late-stage modification of peptides and proteins at cysteine in aqueous buffer with suitably functionalised diaryliodonium salts, furnishing stable thioether-linked synthetic conjugates. The power of this new platform is showcased through the late-stage modification of the affibody zEGFR and the histone protein H2A.},\n bibtype = {article},\n author = {Byrne, S.A. and Bedding, M.J. and Corcilius, L. and Ford, D.J. and Zhong, Y. and Franck, C. and Larance, M. and Mackay, J.P. and Payne, R.J.},\n doi = {10.1039/d1sc03127a},\n journal = {Chemical Science},\n number = {42}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The modification of peptides and proteins has emerged as a powerful means to efficiently prepare high value bioconjugates for a range of applications in chemical biology and for the development of next-generation therapeutics. Herein, we report a novel method for the chemoselective late-stage modification of peptides and proteins at cysteine in aqueous buffer with suitably functionalised diaryliodonium salts, furnishing stable thioether-linked synthetic conjugates. The power of this new platform is showcased through the late-stage modification of the affibody zEGFR and the histone protein H2A.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"counoupas-johansen-stella-nguyen-ferguson-aggarwal-bhattacharyya-grey-etal-asingledosebcgadjuvantedcovid19vaccineprovidessterilisingimmunityagainstsarscov2infection-2021\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A single dose, BCG-adjuvanted COVID-19 vaccine provides sterilising immunity against SARS-CoV-2 infection.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Counoupas,  C.; Johansen,  M.; Stella,  A.; Nguyen,  D.; Ferguson,  A.; Aggarwal,  A.; Bhattacharyya,  N.; Grey,  A.; Hutchings,  O.; Patel,  K.; Hansbro,  P.;  and Triccas,  J.\n</span>\n\n  \n\n</span>\n\n  <i>npj Vaccines</i>, 6(1).  2021.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41541-021-00406-4\"\n     onclick=\"log_download('counoupas-johansen-stella-nguyen-ferguson-aggarwal-bhattacharyya-grey-etal-asingledosebcgadjuvantedcovid19vaccineprovidessterilisingimmunityagainstsarscov2infection-2021', 'http://doi.org/10.1038/s41541-021-00406-4', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/counoupas-johansen-stella-nguyen-ferguson-aggarwal-bhattacharyya-grey-etal-asingledosebcgadjuvantedcovid19vaccineprovidessterilisingimmunityagainstsarscov2infection-2021\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('counoupas-johansen-stella-nguyen-ferguson-aggarwal-bhattacharyya-grey-etal-asingledosebcgadjuvantedcovid19vaccineprovidessterilisingimmunityagainstsarscov2infection-2021')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A single dose, BCG-adjuvanted COVID-19 vaccine provides sterilising immunity against SARS-CoV-2 infection},\n type = {article},\n year = {2021},\n volume = {6},\n id = {5f7017bd-f4d1-30e2-95a1-458fa2083a65},\n created = {2023-01-10T01:43:52.472Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:52.472Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Global control of COVID-19 requires broadly accessible vaccines that are effective against SARS-CoV-2 variants. In this report, we exploit the immunostimulatory properties of bacille Calmette-Guérin (BCG), the existing tuberculosis vaccine, to deliver a vaccination regimen with potent SARS-CoV-2-specific protective immunity. Combination of BCG with a stabilised, trimeric form of SARS-CoV-2 spike antigen promoted rapid development of virus-specific IgG antibodies in the blood of vaccinated mice, that was further augmented by the addition of alum. This vaccine formulation, BCG:CoVac, induced high-titre SARS-CoV-2 neutralising antibodies (NAbs) and Th1-biased cytokine release by vaccine-specific T cells, which correlated with the early emergence of T follicular helper cells in local lymph nodes and heightened levels of antigen-specific plasma B cells after vaccination. Vaccination of K18-hACE2 mice with a single dose of BCG:CoVac almost completely abrogated disease after SARS-CoV-2 challenge, with minimal inflammation and no detectable virus in the lungs of infected animals. Boosting BCG:CoVac-primed mice with a heterologous vaccine further increased SARS-CoV-2-specific antibody responses, which effectively neutralised B.1.1.7 and B.1.351 SARS-CoV-2 variants of concern. These findings demonstrate the potential for BCG-based vaccination to protect against major SARS-CoV-2 variants circulating globally.},\n bibtype = {article},\n author = {Counoupas, C. and Johansen, M.D. and Stella, A.O. and Nguyen, D.H. and Ferguson, A.L. and Aggarwal, A. and Bhattacharyya, N.D. and Grey, A. and Hutchings, O. and Patel, K. and Hansbro, P.M. and Triccas, J.A.},\n doi = {10.1038/s41541-021-00406-4},\n journal = {npj Vaccines},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Global control of COVID-19 requires broadly accessible vaccines that are effective against SARS-CoV-2 variants. In this report, we exploit the immunostimulatory properties of bacille Calmette-Guérin (BCG), the existing tuberculosis vaccine, to deliver a vaccination regimen with potent SARS-CoV-2-specific protective immunity. Combination of BCG with a stabilised, trimeric form of SARS-CoV-2 spike antigen promoted rapid development of virus-specific IgG antibodies in the blood of vaccinated mice, that was further augmented by the addition of alum. This vaccine formulation, BCG:CoVac, induced high-titre SARS-CoV-2 neutralising antibodies (NAbs) and Th1-biased cytokine release by vaccine-specific T cells, which correlated with the early emergence of T follicular helper cells in local lymph nodes and heightened levels of antigen-specific plasma B cells after vaccination. Vaccination of K18-hACE2 mice with a single dose of BCG:CoVac almost completely abrogated disease after SARS-CoV-2 challenge, with minimal inflammation and no detectable virus in the lungs of infected animals. Boosting BCG:CoVac-primed mice with a heterologous vaccine further increased SARS-CoV-2-specific antibody responses, which effectively neutralised B.1.1.7 and B.1.351 SARS-CoV-2 variants of concern. These findings demonstrate the potential for BCG-based vaccination to protect against major SARS-CoV-2 variants circulating globally.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"jindra-mckinstry-nebl-bittova-ren-shaw-phan-lu-etal-purificationofaninsectjuvenilehormonereceptorcomplexenablesinsightsintoitsposttranslationalphosphorylation-2021\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Purification of an insect juvenile hormone receptor complex enables insights into its post-translational phosphorylation.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Jindra,  M.; McKinstry,  W.; Nebl,  T.; Bittova,  L.; Ren,  B.; Shaw,  J.; Phan,  T.; Lu,  L.; Low,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Lovrecz,  G.;  and Hill,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 297(6).  2021.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jbc.2021.101387\"\n     onclick=\"log_download('jindra-mckinstry-nebl-bittova-ren-shaw-phan-lu-etal-purificationofaninsectjuvenilehormonereceptorcomplexenablesinsightsintoitsposttranslationalphosphorylation-2021', 'http://doi.org/10.1016/j.jbc.2021.101387', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/jindra-mckinstry-nebl-bittova-ren-shaw-phan-lu-etal-purificationofaninsectjuvenilehormonereceptorcomplexenablesinsightsintoitsposttranslationalphosphorylation-2021\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('jindra-mckinstry-nebl-bittova-ren-shaw-phan-lu-etal-purificationofaninsectjuvenilehormonereceptorcomplexenablesinsightsintoitsposttranslationalphosphorylation-2021')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Purification of an insect juvenile hormone receptor complex enables insights into its post-translational phosphorylation},\n type = {article},\n year = {2021},\n volume = {297},\n id = {f039ef2e-e6b3-3564-a85c-0dc612c27e87},\n created = {2023-01-10T01:43:53.398Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:53.398Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Juvenile hormone (JH) plays vital roles in insect reproduction, development, and in many aspects of physiology. JH primarily acts at the gene-regulatory level through interaction with an intracellular receptor (JH receptor [JHR]), a ligand-activated complex of transcription factors consisting of the JH-binding protein methoprene-tolerant (MET) and its partner taiman (TAI). Initial studies indicated significance of post-transcriptional phosphorylation, subunit assembly, and nucleocytoplasmic transport of JHR in JH signaling. However, our knowledge of JHR regulation at the protein level remains rudimentary, partly because of the difficulty of obtaining purified and functional JHR proteins. Here, we present a method for high-yield expression and purification of JHR complexes from two insect species, the beetle T. castaneum and the mosquito Aedes aegypti. Recombinant JHR subunits from each species were coexpressed in an insect cell line using a baculovirus system. MET–TAI complexes were purified through affinity chromatography and anion exchange columns to yield proteins capable of binding both the hormonal ligand (JH III) and DNA bearing cognate JH-response elements. We further examined the beetle JHR complex in greater detail. Biochemical analyses and MS confirmed that T. castaneum JHR was a 1:1 heterodimer consisting of MET and Taiman proteins, stabilized by the JHR agonist ligand methoprene. Phosphoproteomics uncovered multiple phosphorylation sites in the MET protein, some of which were induced by methoprene treatment. Finally, we report a functional bipartite nuclear localization signal, straddled by phosphorylated residues, within the disordered C-terminal region of MET. Our present characterization of the recombinant JHR is an initial step toward understanding JHR structure and function.},\n bibtype = {article},\n author = {Jindra, M. and McKinstry, W.J. and Nebl, T. and Bittova, L. and Ren, B. and Shaw, J. and Phan, T. and Lu, L. and Low, J.K.K. and Mackay, J.P. and Lovrecz, G.O. and Hill, R.J.},\n doi = {10.1016/j.jbc.2021.101387},\n journal = {Journal of Biological Chemistry},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Juvenile hormone (JH) plays vital roles in insect reproduction, development, and in many aspects of physiology. JH primarily acts at the gene-regulatory level through interaction with an intracellular receptor (JH receptor [JHR]), a ligand-activated complex of transcription factors consisting of the JH-binding protein methoprene-tolerant (MET) and its partner taiman (TAI). Initial studies indicated significance of post-transcriptional phosphorylation, subunit assembly, and nucleocytoplasmic transport of JHR in JH signaling. However, our knowledge of JHR regulation at the protein level remains rudimentary, partly because of the difficulty of obtaining purified and functional JHR proteins. Here, we present a method for high-yield expression and purification of JHR complexes from two insect species, the beetle T. castaneum and the mosquito Aedes aegypti. Recombinant JHR subunits from each species were coexpressed in an insect cell line using a baculovirus system. MET–TAI complexes were purified through affinity chromatography and anion exchange columns to yield proteins capable of binding both the hormonal ligand (JH III) and DNA bearing cognate JH-response elements. We further examined the beetle JHR complex in greater detail. Biochemical analyses and MS confirmed that T. castaneum JHR was a 1:1 heterodimer consisting of MET and Taiman proteins, stabilized by the JHR agonist ligand methoprene. Phosphoproteomics uncovered multiple phosphorylation sites in the MET protein, some of which were induced by methoprene treatment. Finally, we report a functional bipartite nuclear localization signal, straddled by phosphorylated residues, within the disordered C-terminal region of MET. Our present characterization of the recombinant JHR is an initial step toward understanding JHR structure and function.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"norman-franck-christie-hawkins-patel-ashhurst-aggarwal-low-etal-discoveryofcyclicpeptideligandstothesarscov2spikeproteinusingmrnadisplay-2021\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Discovery of Cyclic Peptide Ligands to the SARS-CoV-2 Spike Protein Using mRNA Display.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Norman,  A.; Franck,  C.; Christie,  M.; Hawkins,  P.; Patel,  K.; Ashhurst,  A.; Aggarwal,  A.; Low,  J.; Siddiquee,  R.; Ashley,  C.; Passioura,  T.;  and Payne,  R.\n</span>\n\n  \n\n</span>\n\n  <i>ACS Central Science</i>, 7(6): 1001-1008.  2021.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/acscentsci.0c01708\"\n     onclick=\"log_download('norman-franck-christie-hawkins-patel-ashhurst-aggarwal-low-etal-discoveryofcyclicpeptideligandstothesarscov2spikeproteinusingmrnadisplay-2021', 'http://doi.org/10.1021/acscentsci.0c01708', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/norman-franck-christie-hawkins-patel-ashhurst-aggarwal-low-etal-discoveryofcyclicpeptideligandstothesarscov2spikeproteinusingmrnadisplay-2021\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>4 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('norman-franck-christie-hawkins-patel-ashhurst-aggarwal-low-etal-discoveryofcyclicpeptideligandstothesarscov2spikeproteinusingmrnadisplay-2021')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Discovery of Cyclic Peptide Ligands to the SARS-CoV-2 Spike Protein Using mRNA Display},\n type = {article},\n year = {2021},\n pages = {1001-1008},\n volume = {7},\n id = {278be5c6-73f1-3344-9bd1-51ffab0169e6},\n created = {2023-01-10T01:43:54.314Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:54.314Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The COVID-19 pandemic, caused by SARS-CoV-2, has led to substantial morbidity, mortality, and disruption globally. Cellular entry of SARS-CoV-2 is mediated by the viral spike protein, and affinity ligands to this surface protein have the potential for applications as antivirals and diagnostic reagents. Here, we describe the affinity selection of cyclic peptide ligands to the SARS-CoV-2 spike protein receptor binding domain (RBD) from three distinct libraries (in excess of a trillion molecules each) by mRNA display. We identified six high affinity molecules with dissociation constants (KD) in the nanomolar range (15-550 nM) to the RBD. The highest affinity ligand could be used as an affinity reagent to detect the spike protein in solution by ELISA, and the cocrystal structure of this molecule bound to the RBD demonstrated that it binds to a cryptic binding site, displacing a β-strand near the C-terminus. Our findings provide key mechanistic insight into the binding of peptide ligands to the SARS-CoV-2 spike RBD, and the ligands discovered in this work may find future use as reagents for diagnostic applications.},\n bibtype = {article},\n author = {Norman, A. and Franck, C. and Christie, M. and Hawkins, P.M.E. and Patel, K. and Ashhurst, A.S. and Aggarwal, A. and Low, J.K.K. and Siddiquee, R. and Ashley, C.L. and Passioura, T. and Payne, R.J.},\n doi = {10.1021/acscentsci.0c01708},\n journal = {ACS Central Science},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The COVID-19 pandemic, caused by SARS-CoV-2, has led to substantial morbidity, mortality, and disruption globally. Cellular entry of SARS-CoV-2 is mediated by the viral spike protein, and affinity ligands to this surface protein have the potential for applications as antivirals and diagnostic reagents. Here, we describe the affinity selection of cyclic peptide ligands to the SARS-CoV-2 spike protein receptor binding domain (RBD) from three distinct libraries (in excess of a trillion molecules each) by mRNA display. We identified six high affinity molecules with dissociation constants (KD) in the nanomolar range (15-550 nM) to the RBD. The highest affinity ligand could be used as an affinity reagent to detect the spike protein in solution by ELISA, and the cocrystal structure of this molecule bound to the RBD demonstrated that it binds to a cryptic binding site, displacing a β-strand near the C-terminus. Our findings provide key mechanistic insight into the binding of peptide ligands to the SARS-CoV-2 spike RBD, and the ligands discovered in this work may find future use as reagents for diagnostic applications.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2020\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2020')\"\n      onkeypress=\"toggleGroup('group_2020')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2020</span>\n      <span class=\"bibbase_group_count\">\n        \n        (15)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.org/10.1038/s41436-019-0747-z\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020', 'http://doi.org/10.1038/s41436-019-0747-z', event);\">\n    GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Shieh,  C.; Jones,  N.; Vanle,  B.; Au,  M.; Huang,  A., Y.; Silva,  A., P., G.; Lee,  H.; Douine,  E., D.; Otero,  M., G.; Choi,  A.; Grand,  K.; Taff,  I., P.; Delgado,  M., R.; Hajianpour,  M., J.; Seeley,  A.; Rohena,  L.; Vernon,  H.; Gripp,  K., W.; Vergano,  S., A.; Mahida,  S.; Naidu,  S.; Sousa,  A., B.; Wain,  K., E.; Challman,  T., D.; Beek,  G.; Basel,  D.; Ranells,  J.; Smith,  R.; Yusupov,  R.; Freckmann,  M.; Ohden,  L.; Davis-Keppen,  L.; Chitayat,  D.; Dowling,  J., J.; Finkel,  R.; Dauber,  A.; Spillmann,  R.; Pena,  L., D., M.; Metcalfe,  K.; Splitt,  M.; Lachlan,  K.; McKee,  S., A.; Hurst,  J.; Fitzpatrick,  D., R.; Morton,  J., E., V.; Cox,  H.; Venkateswaran,  S.; Young,  J., I.; Marsh,  E., D.; Nelson,  S., F.; Martinez,  J., A.; Graham,  J., M.; Kini,  U.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Pierson,  T., M.\n</span>\n\n  \n\n</span>\n\n  <i>Genetics in Medicine</i>, 22(5): 878-888. 5 2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.org/10.1038/s41436-019-0747-z\"\n     onclick=\"log_download('shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020', 'http://doi.org/10.1038/s41436-019-0747-z', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41436-019-0747-z\"\n     onclick=\"log_download('shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020', 'http://doi.org/10.1038/s41436-019-0747-z', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder},\n type = {article},\n year = {2020},\n keywords = {GATAD2B,NuRD complex,apraxia of speech,chromati},\n pages = {878-888},\n volume = {22},\n websites = {http://doi.org/10.1038/s41436-019-0747-z},\n month = {5},\n day = {17},\n id = {6308af4d-95ba-3986-b5f1-b8362afa6d89},\n created = {2020-12-17T05:29:53.237Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:53.237Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Shieh2020},\n source_type = {article},\n private_publication = {false},\n abstract = {Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.},\n bibtype = {article},\n author = {Shieh, Christine and Jones, Natasha and Vanle, Brigitte and Au, Margaret and Huang, Alden Y. and Silva, Ana P. G. and Lee, Hane and Douine, Emilie D. and Otero, Maria G. and Choi, Andrew and Grand, Katheryn and Taff, Ingrid P. and Delgado, Mauricio R. and Hajianpour, M. J. and Seeley, Andrea and Rohena, Luis and Vernon, Hilary and Gripp, Karen W. and Vergano, Samantha A. and Mahida, Sonal and Naidu, Sakkubai and Sousa, Ana Berta and Wain, Karen E. and Challman, Thomas D. and Beek, Geoffrey and Basel, Donald and Ranells, Judith and Smith, Rosemarie and Yusupov, Roman and Freckmann, Mary-Louise and Ohden, Lisa and Davis-Keppen, Laura and Chitayat, David and Dowling, James J. and Finkel, Richard and Dauber, Andrew and Spillmann, Rebecca and Pena, Loren D. M. and Metcalfe, Kay and Splitt, Miranda and Lachlan, Katherine and McKee, Shane A. and Hurst, Jane and Fitzpatrick, David R. and Morton, Jenny E. V. and Cox, Helen and Venkateswaran, Sunita and Young, Juan I. and Marsh, Eric D. and Nelson, Stanley F. and Martinez, Julian A. and Graham, John M. and Kini, Usha and Mackay, Joel P. and Pierson, Tyler Mark},\n doi = {10.1038/s41436-019-0747-z},\n journal = {Genetics in Medicine},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/31622516\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020', 'http://www.ncbi.nlm.nih.gov/pubmed/31622516', event);\">\n    Peppy: A virtual reality environment for exploring the principles of polypeptide structure.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Doak,  D., G.; Denyer,  G., S.; Gerrard,  J., A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Allison,  J., R.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 29(1): 157-168. 1 2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/31622516\"\n     onclick=\"log_download('doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020', 'http://www.ncbi.nlm.nih.gov/pubmed/31622516', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Peppy: A virtual reality environment for exploring the principles of polypeptide structure [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.3752\"\n     onclick=\"log_download('doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020', 'http://doi.org/10.1002/pro.3752', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Peppy: A virtual reality environment for exploring the principles of polypeptide structure},\n type = {article},\n year = {2020},\n keywords = {polypeptide,protein,secondary structure,teaching,undergraduate,virtual reality},\n pages = {157-168},\n volume = {29},\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/31622516},\n month = {1},\n day = {11},\n id = {fe01e53b-d05d-3939-8395-e8da99c6004e},\n created = {2020-12-17T05:29:55.556Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:55.556Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Doak2020},\n source_type = {article},\n private_publication = {false},\n abstract = {A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.},\n bibtype = {article},\n author = {Doak, David G. and Denyer, Gareth S. and Gerrard, Juliet A. and Mackay, Joel P. and Allison, Jane R.},\n doi = {10.1002/pro.3752},\n journal = {Protein Science},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Cyclic peptides can engage a single binding pocket through highly divergent modes.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Patel,  K.; Walport,  L.; Walshe,  J.; Solomon,  P.; Low,  J.; Tran,  D.; Mouradian,  K.; Silva,  A.; Wilkinson-White,  L.; Norman,  A.; Franck,  C.; Matthews,  J.; Mitchell Guss,  J.; Payne,  R.; Passioura,  T.; Suga,  H.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 117(43).  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.2003086117\"\n     onclick=\"log_download('patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020', 'http://doi.org/10.1073/pnas.2003086117', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Cyclic peptides can engage a single binding pocket through highly divergent modes},\n type = {article},\n year = {2020},\n keywords = {BET bromodomain inhibition,BRD3,BRD4,De novo cyclic peptides,Structural biology},\n volume = {117},\n id = {6636c6a0-e2f0-3aea-af8c-7865a5f17fcf},\n created = {2020-12-17T07:26:41.435Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T07:26:41.435Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {© 2020 National Academy of Sciences. All rights reserved. Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.},\n bibtype = {article},\n author = {Patel, K. and Walport, L.J. and Walshe, J.L. and Solomon, P.D. and Low, J.K.K. and Tran, D.H. and Mouradian, K.S. and Silva, A.P.G. and Wilkinson-White, L. and Norman, A. and Franck, C. and Matthews, J.M. and Mitchell Guss, J. and Payne, R.J. and Passioura, T. and Suga, H. and Mackay, J.P.},\n doi = {10.1073/pnas.2003086117},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {43}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  © 2020 National Academy of Sciences. All rights reserved. Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Shieh,  C.; Jones,  N.; Vanle,  B.; Au,  M.; Huang,  A.; Silva,  A.; Lee,  H.; Douine,  E.; Otero,  M.; Choi,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Pierson,  T.\n</span>\n\n  \n\n</span>\n\n  <i>Genetics in Medicine</i>, 22(5): 878-888.  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41436-019-0747-z\"\n     onclick=\"log_download('shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020', 'http://doi.org/10.1038/s41436-019-0747-z', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('shieh-jones-vanle-au-huang-silva-lee-douine-etal-gatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisorder-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder},\n type = {article},\n year = {2020},\n pages = {878-888},\n volume = {22},\n id = {85308366-aa04-3a25-93ec-e3edc0ad78cc},\n created = {2023-01-10T01:43:55.280Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:55.280Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.},\n bibtype = {article},\n author = {Shieh, C. and Jones, N. and Vanle, B. and Au, M. and Huang, A.Y. and Silva, A.P.G. and Lee, H. and Douine, E.D. and Otero, M.G. and Choi, A. and Mackay, J.P. and Pierson, T.M.},\n doi = {10.1038/s41436-019-0747-z},\n journal = {Genetics in Medicine},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Purpose: Determination of genotypic/phenotypic features of GATAD2B-associated neurodevelopmental disorder(GAND). Methods: Fifty GAND subjects were evaluated to determine consistentgenotypic/phenotypic features. Immunoprecipitation assays utilizing in vitrotranscription–translation products were used to evaluate GATAD2B missensevariants’ ability to interact with binding partners within the nucleosomeremodeling and deacetylase (NuRD) complex. Results: Subjects had clinical findings that included macrocephaly,hypotonia, intellectual disability, neonatal feeding issues, polyhydramnios,apraxia of speech, epilepsy, and bicuspid aortic valves. Forty-one novelGATAD2B variants were identified withmultiple variant types (nonsense, truncating frameshift, splice-site variants,deletions, and missense). Seven subjects were identified with missense variantsthat localized within two conserved region domains (CR1 or CR2) of the GATAD2Bprotein. Immunoprecipitation assays revealed several of these missense variantsdisrupted GATAD2B interactions with its NuRD complex binding partners. Conclusions: A consistent GAND phenotype was caused by a range of geneticvariants in GATAD2B that includeloss-of-function and missense subtypes. Missense variants were present inconserved region domains that disrupted assembly of NuRD complex proteins.GAND’s clinical phenotype had substantial clinical overlap with other disordersassociated with the NuRD complex that involve CHD3 and CHD4, with clinicalfeatures of hypotonia, intellectual disability, cardiac defects, childhoodapraxia of speech, and macrocephaly.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"werner-wang-hamagami-hsu-yano-stonestrom-behera-zhong-etal-correctioncomparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystemappliedoptics202029518981914doi101074jbcra119010679-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Correction: Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system.(Applied Optics(2020)295(1898–1914)Doi: 10.1074/jbc.RA119.010679).\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Werner,  M.; Wang,  H.; Hamagami,  N.; Hsu,  S.; Yano,  J.; Stonestrom,  A.; Behera,  V.; Zhong,  Y.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Blobel,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 295(18): 6251.  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.AAC120.013771\"\n     onclick=\"log_download('werner-wang-hamagami-hsu-yano-stonestrom-behera-zhong-etal-correctioncomparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystemappliedoptics202029518981914doi101074jbcra119010679-2020', 'http://doi.org/10.1074/jbc.AAC120.013771', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/werner-wang-hamagami-hsu-yano-stonestrom-behera-zhong-etal-correctioncomparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystemappliedoptics202029518981914doi101074jbcra119010679-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('werner-wang-hamagami-hsu-yano-stonestrom-behera-zhong-etal-correctioncomparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystemappliedoptics202029518981914doi101074jbcra119010679-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Correction: Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system.(Applied Optics(2020)295(1898–1914)Doi: 10.1074/jbc.RA119.010679)},\n type = {article},\n year = {2020},\n pages = {6251},\n volume = {295},\n id = {9fc40b26-4df6-3126-8c8e-92fc8825b167},\n created = {2023-01-10T01:43:56.242Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:56.242Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Yichen Zhong&#x27;s name was misspelled. The correct spelling is shown above.},\n bibtype = {article},\n author = {Werner, M.T. and Wang, H. and Hamagami, N. and Hsu, S.C. and Yano, J.A. and Stonestrom, A.J. and Behera, V. and Zhong, Y. and MacKay, J.P. and Blobel, G.A.},\n doi = {10.1074/jbc.AAC120.013771},\n journal = {Journal of Biological Chemistry},\n number = {18}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Yichen Zhong's name was misspelled. The correct spelling is shown above.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Cyclic peptides can engage a single binding pocket through highly divergent modes.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Patel,  K.; Walport,  L.; Walshe,  J.; Solomon,  P.; Low,  J.; Tran,  D.; Mouradian,  K.; Silva,  A.; Wilkinson-White,  L.; Norman,  A.; Suga,  H.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 117(43): 26728-26738.  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.2003086117\"\n     onclick=\"log_download('patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020', 'http://doi.org/10.1073/pnas.2003086117', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('patel-walport-walshe-solomon-low-tran-mouradian-silva-etal-cyclicpeptidescanengageasinglebindingpocketthroughhighlydivergentmodes-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Cyclic peptides can engage a single binding pocket through highly divergent modes},\n type = {article},\n year = {2020},\n pages = {26728-26738},\n volume = {117},\n id = {9d68a322-e2a2-3b30-b21a-47d62f211339},\n created = {2023-01-10T01:43:57.164Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:57.164Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.},\n bibtype = {article},\n author = {Patel, K. and Walport, L.J. and Walshe, J.L. and Solomon, P.D. and Low, J.K.K. and Tran, D.H. and Mouradian, K.S. and Silva, A.P.G. and Wilkinson-White, L. and Norman, A. and Suga, H. and Mackay, J.P.},\n doi = {10.1073/pnas.2003086117},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {43}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Cyclic peptide library screening technologies show immense promise for identifying drug leads and chemical probes for challenging targets. However, the structural and functional diversity encoded within such libraries is largely undefined. We have systematically profiled the affinity, selectivity, and structural features of library-derived cyclic peptides selected to recognize three closely related targets: the acetyllysine-binding bromodomain proteins BRD2, -3, and -4. We report affinities as low as 100 pM and specificities of up to 106-fold. Crystal structures of 13 peptide–bromodomain complexes reveal remarkable diversity in both structure and binding mode, including both α-helical and β-sheet structures as well as bivalent binding modes. The peptides can also exhibit a high degree of structural preorganization. Our data demonstrate the enormous potential within these libraries to provide diverse binding modes against a single target, which underpins their capacity to yield highly potent and selective ligands.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"franck-foster-johansenleete-chowdhury-cielesh-bhusal-mackay-larance-etal-semisynthesisofanevasinfromticksalivarevealsacriticalroleoftyrosinesulfationforchemokinebindingandinhibition-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Semisynthesis of an evasin from tick saliva reveals a critical role of tyrosine sulfation for chemokine binding and inhibition.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Franck,  C.; Foster,  S.; Johansen-Leete,  J.; Chowdhury,  S.; Cielesh,  M.; Bhusal,  R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Larance,  M.; Stone,  M.;  and Payne,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 117(23): 12657-12664.  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.2000605117\"\n     onclick=\"log_download('franck-foster-johansenleete-chowdhury-cielesh-bhusal-mackay-larance-etal-semisynthesisofanevasinfromticksalivarevealsacriticalroleoftyrosinesulfationforchemokinebindingandinhibition-2020', 'http://doi.org/10.1073/pnas.2000605117', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/franck-foster-johansenleete-chowdhury-cielesh-bhusal-mackay-larance-etal-semisynthesisofanevasinfromticksalivarevealsacriticalroleoftyrosinesulfationforchemokinebindingandinhibition-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>4 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('franck-foster-johansenleete-chowdhury-cielesh-bhusal-mackay-larance-etal-semisynthesisofanevasinfromticksalivarevealsacriticalroleoftyrosinesulfationforchemokinebindingandinhibition-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Semisynthesis of an evasin from tick saliva reveals a critical role of tyrosine sulfation for chemokine binding and inhibition},\n type = {article},\n year = {2020},\n pages = {12657-12664},\n volume = {117},\n id = {b8c84666-fc07-3ad8-aaf9-d251ef3b0ee8},\n created = {2023-01-10T01:43:58.083Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:58.083Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Blood-feeding arthropods produce antiinflammatory salivary proteins called evasins that function through inhibition of chemokine-receptor signaling in the host. Herein, we show that the evasin ACA-01 from the Amblyomma cajennense tick can be posttranslationally sulfated at two tyrosine residues, albeit as a mixture of sulfated variants. Homogenously sulfated variants of the proteins were efficiently assembled via a semisynthetic native chemical ligation strategy. Sulfation significantly improved the binding affinity of ACA-01 for a range of proinflammatory chemokines and enhanced the ability of ACA-01 to inhibit chemokine signaling through cognate receptors. Comparisons of evasin sequences and structural data suggest that tyrosine sulfation serves as a receptor mimetic strategy for recognizing and suppressing the proinflammatory activity of a wide variety of mammalian chemokines. As such, the incorporation of this posttranslational modification (PTM) or mimics thereof into evasins may provide a strategy to optimize tick salivary proteins for antiinflammatory applications.},\n bibtype = {article},\n author = {Franck, C. and Foster, S.R. and Johansen-Leete, J. and Chowdhury, S. and Cielesh, M. and Bhusal, R.P. and Mackay, J.P. and Larance, M. and Stone, M.J. and Payne, R.J.},\n doi = {10.1073/pnas.2000605117},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {23}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Blood-feeding arthropods produce antiinflammatory salivary proteins called evasins that function through inhibition of chemokine-receptor signaling in the host. Herein, we show that the evasin ACA-01 from the Amblyomma cajennense tick can be posttranslationally sulfated at two tyrosine residues, albeit as a mixture of sulfated variants. Homogenously sulfated variants of the proteins were efficiently assembled via a semisynthetic native chemical ligation strategy. Sulfation significantly improved the binding affinity of ACA-01 for a range of proinflammatory chemokines and enhanced the ability of ACA-01 to inhibit chemokine signaling through cognate receptors. Comparisons of evasin sequences and structural data suggest that tyrosine sulfation serves as a receptor mimetic strategy for recognizing and suppressing the proinflammatory activity of a wide variety of mammalian chemokines. As such, the incorporation of this posttranslational modification (PTM) or mimics thereof into evasins may provide a strategy to optimize tick salivary proteins for antiinflammatory applications.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"xi-difazio-nadvi-patel-xiang-zhang-deshpande-low-etal-anovelpurificationprocedureforactiverecombinanthumandpp4andtheinabilityofdpp4tobindsarscov2-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A Novel Purification Procedure for Active Recombinant Human DPP4 and the Inability of DPP4 to Bind SARS-CoV-2.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Xi,  C.; Di Fazio,  A.; Nadvi,  N.; Patel,  K.; Xiang,  M.; Zhang,  H.; Deshpande,  C.; Low,  J.; Wang,  X.; Chen,  Y.; Church,  W.;  and Gorrell,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Molecules</i>, 25(22).  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.3390/MOLECULES25225392\"\n     onclick=\"log_download('xi-difazio-nadvi-patel-xiang-zhang-deshpande-low-etal-anovelpurificationprocedureforactiverecombinanthumandpp4andtheinabilityofdpp4tobindsarscov2-2020', 'http://doi.org/10.3390/MOLECULES25225392', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/xi-difazio-nadvi-patel-xiang-zhang-deshpande-low-etal-anovelpurificationprocedureforactiverecombinanthumandpp4andtheinabilityofdpp4tobindsarscov2-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('xi-difazio-nadvi-patel-xiang-zhang-deshpande-low-etal-anovelpurificationprocedureforactiverecombinanthumandpp4andtheinabilityofdpp4tobindsarscov2-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A Novel Purification Procedure for Active Recombinant Human DPP4 and the Inability of DPP4 to Bind SARS-CoV-2},\n type = {article},\n year = {2020},\n volume = {25},\n id = {016baf4e-9b3e-39c6-87c2-be2e11c5cbaa},\n created = {2023-01-10T01:43:59.005Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:59.005Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Proteases catalyse irreversible posttranslational modifications that often alter a biological function of the substrate. The protease dipeptidyl peptidase 4 (DPP4) is a pharmacological target in type 2 diabetes therapy primarily because it inactivates glucagon-like protein-1. DPP4 also has roles in steatosis, insulin resistance, cancers and inflammatory and fibrotic diseases. In addition, DPP4 binds to the spike protein of the MERS virus, causing it to be the human cell surface receptor for that virus. DPP4 has been identified as a potential binding target of SARS-CoV-2 spike protein, so this question requires experimental investigation. Understanding protein structure and function requires reliable protocols for production and purification. We developed such strategies for baculovirus generated soluble recombinant human DPP4 (residues 29–766) produced in insect cells. Purification used differential ammonium sulphate precipitation, hydrophobic interaction chromatography, dye affinity chromatography in series with immobilised metal affinity chromatography, and ion-exchange chromatography. The binding affinities of DPP4 to the SARS-CoV-2 full-length spike protein and its receptor-binding domain (RBD) were measured using surface plasmon resonance and ELISA. This optimised DPP4 purification procedure yielded 1 to 1.8 mg of pure fully active soluble DPP4 protein per litre of insect cell culture with specific activity &gt;30 U/mg, indicative of high purity. No specific binding between DPP4 and CoV-2 spike protein was detected by surface plasmon resonance or ELISA. In summary, a procedure for high purity high yield soluble human DPP4 was achieved and used to show that, unlike MERS, SARS-CoV-2 does not bind human DPP4.},\n bibtype = {article},\n author = {Xi, C.R. and Di Fazio, A. and Nadvi, N.A. and Patel, K. and Xiang, M.S.W. and Zhang, H.E. and Deshpande, C. and Low, J.K.K. and Wang, X.T. and Chen, Y. and Church, W.B. and Gorrell, M.D.},\n doi = {10.3390/MOLECULES25225392},\n journal = {Molecules},\n number = {22}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Proteases catalyse irreversible posttranslational modifications that often alter a biological function of the substrate. The protease dipeptidyl peptidase 4 (DPP4) is a pharmacological target in type 2 diabetes therapy primarily because it inactivates glucagon-like protein-1. DPP4 also has roles in steatosis, insulin resistance, cancers and inflammatory and fibrotic diseases. In addition, DPP4 binds to the spike protein of the MERS virus, causing it to be the human cell surface receptor for that virus. DPP4 has been identified as a potential binding target of SARS-CoV-2 spike protein, so this question requires experimental investigation. Understanding protein structure and function requires reliable protocols for production and purification. We developed such strategies for baculovirus generated soluble recombinant human DPP4 (residues 29–766) produced in insect cells. Purification used differential ammonium sulphate precipitation, hydrophobic interaction chromatography, dye affinity chromatography in series with immobilised metal affinity chromatography, and ion-exchange chromatography. The binding affinities of DPP4 to the SARS-CoV-2 full-length spike protein and its receptor-binding domain (RBD) were measured using surface plasmon resonance and ELISA. This optimised DPP4 purification procedure yielded 1 to 1.8 mg of pure fully active soluble DPP4 protein per litre of insect cell culture with specific activity >30 U/mg, indicative of high purity. No specific binding between DPP4 and CoV-2 spike protein was detected by surface plasmon resonance or ELISA. In summary, a procedure for high purity high yield soluble human DPP4 was achieved and used to show that, unlike MERS, SARS-CoV-2 does not bind human DPP4.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"zhong-paudel-ryan-low-franck-patel-bedward-torrado-etal-chd4slidesnucleosomesbydecouplingentryandexitsidednatranslocation-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  CHD4 slides nucleosomes by decoupling entry- and exit-side DNA translocation.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Zhong,  Y.; Paudel,  B.; Ryan,  D.; Low,  J.; Franck,  C.; Patel,  K.; Bedward,  M.; Torrado,  M.; Payne,  R.; van Oijen,  A.; van Oijen,  A.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Nature Communications</i>, 11(1).  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41467-020-15183-2\"\n     onclick=\"log_download('zhong-paudel-ryan-low-franck-patel-bedward-torrado-etal-chd4slidesnucleosomesbydecouplingentryandexitsidednatranslocation-2020', 'http://doi.org/10.1038/s41467-020-15183-2', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/zhong-paudel-ryan-low-franck-patel-bedward-torrado-etal-chd4slidesnucleosomesbydecouplingentryandexitsidednatranslocation-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('zhong-paudel-ryan-low-franck-patel-bedward-torrado-etal-chd4slidesnucleosomesbydecouplingentryandexitsidednatranslocation-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {CHD4 slides nucleosomes by decoupling entry- and exit-side DNA translocation},\n type = {article},\n year = {2020},\n volume = {11},\n id = {5dc9783b-933a-3c63-9cd9-b4ae4e6fd62e},\n created = {2023-01-10T01:43:59.935Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:43:59.935Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Chromatin remodellers hydrolyse ATP to move nucleosomal DNA against histone octamers. The mechanism, however, is only partially resolved, and it is unclear if it is conserved among the four remodeller families. Here we use single-molecule assays to examine the mechanism of action of CHD4, which is part of the least well understood family. We demonstrate that the binding energy for CHD4-nucleosome complex formation—even in the absence of nucleotide—triggers significant conformational changes in DNA at the entry side, effectively priming the system for remodelling. During remodelling, flanking DNA enters the nucleosome in a continuous, gradual manner but exits in concerted 4–6 base-pair steps. This decoupling of entry- and exit-side translocation suggests that ATP-driven movement of entry-side DNA builds up strain inside the nucleosome that is subsequently released at the exit side by DNA expulsion. Based on our work and previous studies, we propose a mechanism for nucleosome sliding.},\n bibtype = {article},\n author = {Zhong, Y. and Paudel, B.P. and Ryan, D.P. and Low, J.K.K. and Franck, C. and Patel, K. and Bedward, M.J. and Torrado, M. and Payne, R.J. and van Oijen, A.M. and van Oijen, A.M. and Mackay, J.P.},\n doi = {10.1038/s41467-020-15183-2},\n journal = {Nature Communications},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chromatin remodellers hydrolyse ATP to move nucleosomal DNA against histone octamers. The mechanism, however, is only partially resolved, and it is unclear if it is conserved among the four remodeller families. Here we use single-molecule assays to examine the mechanism of action of CHD4, which is part of the least well understood family. We demonstrate that the binding energy for CHD4-nucleosome complex formation—even in the absence of nucleotide—triggers significant conformational changes in DNA at the entry side, effectively priming the system for remodelling. During remodelling, flanking DNA enters the nucleosome in a continuous, gradual manner but exits in concerted 4–6 base-pair steps. This decoupling of entry- and exit-side translocation suggests that ATP-driven movement of entry-side DNA builds up strain inside the nucleosome that is subsequently released at the exit side by DNA expulsion. Based on our work and previous studies, we propose a mechanism for nucleosome sliding.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"latham-torrado-atto-walshe-wilson-guss-mackay-tristram-etal-ahemebindingproteinproducedbyhaemophilushaemolyticusinhibitsnontypeablehaemophilusinfluenzae-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A heme-binding protein produced by Haemophilus haemolyticus inhibits non-typeable Haemophilus influenzae.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Latham,  R.; Torrado,  M.; Atto,  B.; Walshe,  J.; Wilson,  R.; Guss,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Tristram,  S.;  and Gell,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Molecular Microbiology</i>, 113(2): 381-398.  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1111/mmi.14426\"\n     onclick=\"log_download('latham-torrado-atto-walshe-wilson-guss-mackay-tristram-etal-ahemebindingproteinproducedbyhaemophilushaemolyticusinhibitsnontypeablehaemophilusinfluenzae-2020', 'http://doi.org/10.1111/mmi.14426', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/latham-torrado-atto-walshe-wilson-guss-mackay-tristram-etal-ahemebindingproteinproducedbyhaemophilushaemolyticusinhibitsnontypeablehaemophilusinfluenzae-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>4 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('latham-torrado-atto-walshe-wilson-guss-mackay-tristram-etal-ahemebindingproteinproducedbyhaemophilushaemolyticusinhibitsnontypeablehaemophilusinfluenzae-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A heme-binding protein produced by Haemophilus haemolyticus inhibits non-typeable Haemophilus influenzae},\n type = {article},\n year = {2020},\n pages = {381-398},\n volume = {113},\n id = {9a9059a4-c040-3b6b-8ed2-e2bf25cedaa5},\n created = {2023-01-10T01:44:00.856Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:00.856Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Commensal bacteria serve as an important line of defense against colonisation by opportunisitic pathogens, but the underlying molecular mechanisms remain poorly explored. Here, we show that strains of a commensal bacterium, Haemophilus haemolyticus, make hemophilin, a heme-binding protein that inhibits growth of the opportunistic pathogen, non-typeable Haemophilus influenzae (NTHi) in culture. We purified the NTHi-inhibitory protein from H. haemolyticus and identified the hemophilin gene using proteomics and a gene knockout. An x-ray crystal structure of recombinant hemophilin shows that the protein does not belong to any of the known heme-binding protein folds, suggesting that it evolved independently. Biochemical characterisation shows that heme can be captured in the ferrous or ferric state, and with a variety of small heme-ligands bound, suggesting that hemophilin could function under a range of physiological conditions. Hemophilin knockout bacteria show a limited capacity to utilise free heme for growth. Our data suggest that hemophilin is a hemophore and that inhibition of NTHi occurs by heme starvation, raising the possibility that competition from hemophilin-producing H. haemolyticus could antagonise NTHi colonisation in the respiratory tract.},\n bibtype = {article},\n author = {Latham, R.D. and Torrado, M. and Atto, B. and Walshe, J.L. and Wilson, R. and Guss, J.M. and Mackay, J.P. and Tristram, S. and Gell, D.A.},\n doi = {10.1111/mmi.14426},\n journal = {Molecular Microbiology},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Commensal bacteria serve as an important line of defense against colonisation by opportunisitic pathogens, but the underlying molecular mechanisms remain poorly explored. Here, we show that strains of a commensal bacterium, Haemophilus haemolyticus, make hemophilin, a heme-binding protein that inhibits growth of the opportunistic pathogen, non-typeable Haemophilus influenzae (NTHi) in culture. We purified the NTHi-inhibitory protein from H. haemolyticus and identified the hemophilin gene using proteomics and a gene knockout. An x-ray crystal structure of recombinant hemophilin shows that the protein does not belong to any of the known heme-binding protein folds, suggesting that it evolved independently. Biochemical characterisation shows that heme can be captured in the ferrous or ferric state, and with a variety of small heme-ligands bound, suggesting that hemophilin could function under a range of physiological conditions. Hemophilin knockout bacteria show a limited capacity to utilise free heme for growth. Our data suggest that hemophilin is a hemophore and that inhibition of NTHi occurs by heme starvation, raising the possibility that competition from hemophilin-producing H. haemolyticus could antagonise NTHi colonisation in the respiratory tract.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"werner-wang-hamagami-hsu-yano-stonestrom-behera-zong-etal-comparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystem-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Werner,  M.; Wang,  H.; Hamagami,  N.; Hsu,  S.; Yano,  J.; Stonestrom,  A.; Behera,  V.; Zong,  Y.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Blobel,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 295(7): 1898-1914.  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.RA119.010679\"\n     onclick=\"log_download('werner-wang-hamagami-hsu-yano-stonestrom-behera-zong-etal-comparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystem-2020', 'http://doi.org/10.1074/jbc.RA119.010679', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/werner-wang-hamagami-hsu-yano-stonestrom-behera-zong-etal-comparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystem-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('werner-wang-hamagami-hsu-yano-stonestrom-behera-zong-etal-comparativestructurefunctionanalysisofbromodomainandextraterminalmotifbetproteinsinagenecomplementationsystem-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Comparative structure-function analysis of bromodomain and extraterminal motif (BET) proteins in a gene-complementation system},\n type = {article},\n year = {2020},\n pages = {1898-1914},\n volume = {295},\n id = {53a92f38-e8f3-3c7c-9fcf-6b8b0bb9521a},\n created = {2023-01-10T01:44:01.787Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:01.787Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The widely expressed bromodomain and extraterminal motif (BET) proteins bromodomain-containing protein 2 (BRD2), BRD3, and BRD4 are multifunctional transcriptional regulators that bind acetylated chromatin via their conserved tandem bromodomains. Small molecules that target BET bromodomains are being tested for various diseases but typically do not discern between BET family members. Genomic distributions and protein partners of BET proteins have been described, but the basis for differences in BET protein function within a given lineage remains unclear. By establishing a gene knockout-rescue system in a Brd2-null erythroblast cell line, here we compared a series of mutant and chimeric BET proteins for their ability to modulate cell growth, differentiation, and gene expression. We found that the BET N-terminal halves bearing the bromodomains convey marked differences in protein stability but do not account for specificity in BET protein function. Instead, when BET proteins were expressed at comparable levels, their specificity was largely determined by the C-terminal half. Remarkably, a chimeric BET protein comprising the N-terminal half of the structurally similar short BRD4 isoform (BRD4S) and the C-terminal half of BRD2 functioned similarly to intact BRD2. We traced part of the BRD2-specific activity to a previously uncharacterized short segment predicted to harbor a coiled-coil (CC) domain. Deleting the CC segment impaired BRD2&#x27;s ability to restore growth and differentiation, and the CC region functioned in conjunction with the adjacent ET domain to impart BRD2-like activity onto BRD4S. In summary, our results identify distinct BET protein domains that regulate protein turnover and biological activities.},\n bibtype = {article},\n author = {Werner, M.T. and Wang, H. and Hamagami, N. and Hsu, S.C. and Yano, J.A. and Stonestrom, A.J. and Behera, V. and Zong, Y. and Mackay, J.P. and Blobel, G.A.},\n doi = {10.1074/jbc.RA119.010679},\n journal = {Journal of Biological Chemistry},\n number = {7}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The widely expressed bromodomain and extraterminal motif (BET) proteins bromodomain-containing protein 2 (BRD2), BRD3, and BRD4 are multifunctional transcriptional regulators that bind acetylated chromatin via their conserved tandem bromodomains. Small molecules that target BET bromodomains are being tested for various diseases but typically do not discern between BET family members. Genomic distributions and protein partners of BET proteins have been described, but the basis for differences in BET protein function within a given lineage remains unclear. By establishing a gene knockout-rescue system in a Brd2-null erythroblast cell line, here we compared a series of mutant and chimeric BET proteins for their ability to modulate cell growth, differentiation, and gene expression. We found that the BET N-terminal halves bearing the bromodomains convey marked differences in protein stability but do not account for specificity in BET protein function. Instead, when BET proteins were expressed at comparable levels, their specificity was largely determined by the C-terminal half. Remarkably, a chimeric BET protein comprising the N-terminal half of the structurally similar short BRD4 isoform (BRD4S) and the C-terminal half of BRD2 functioned similarly to intact BRD2. We traced part of the BRD2-specific activity to a previously uncharacterized short segment predicted to harbor a coiled-coil (CC) domain. Deleting the CC segment impaired BRD2's ability to restore growth and differentiation, and the CC region functioned in conjunction with the adjacent ET domain to impart BRD2-like activity onto BRD4S. In summary, our results identify distinct BET protein domains that regulate protein turnover and biological activities.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"li-alhendi-yeh-elahy-santiago-deshpande-wu-chan-etal-thermostablesmallmoleculeinhibitorofangiogenesisandvascularpermeabilitythatsuppressesaperkfosbfosbvcam1axis-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Thermostable small-molecule inhibitor of angiogenesis and vascular permeability that suppresses a pERK-FosB/ΔFosB-VCAM-1 axis.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Li,  Y.; Alhendi,  A.; Yeh,  M.; Elahy,  M.; Santiago,  F.; Deshpande,  N.; Wu,  B.; Chan,  E.; Inam,  S.; Prado-Lourenco,  L.; Marcuccio,  S.;  and Khachigian,  L.\n</span>\n\n  \n\n</span>\n\n  <i>Science Advances</i>, 6(31).  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1126/sciadv.aaz7815\"\n     onclick=\"log_download('li-alhendi-yeh-elahy-santiago-deshpande-wu-chan-etal-thermostablesmallmoleculeinhibitorofangiogenesisandvascularpermeabilitythatsuppressesaperkfosbfosbvcam1axis-2020', 'http://doi.org/10.1126/sciadv.aaz7815', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/li-alhendi-yeh-elahy-santiago-deshpande-wu-chan-etal-thermostablesmallmoleculeinhibitorofangiogenesisandvascularpermeabilitythatsuppressesaperkfosbfosbvcam1axis-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('li-alhendi-yeh-elahy-santiago-deshpande-wu-chan-etal-thermostablesmallmoleculeinhibitorofangiogenesisandvascularpermeabilitythatsuppressesaperkfosbfosbvcam1axis-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Thermostable small-molecule inhibitor of angiogenesis and vascular permeability that suppresses a pERK-FosB/ΔFosB-VCAM-1 axis},\n type = {article},\n year = {2020},\n volume = {6},\n id = {f819966b-5abf-3066-bb8c-9914a970f868},\n created = {2023-01-10T01:44:02.733Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:02.733Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Vascular permeability and angiogenesis underpin neovascular age-related macular degeneration and diabetic retinopathy. While anti-VEGF therapies are widely used clinically, many patients do not respond optimally, or at all, and small-molecule therapies are lacking. Here, we identified a dibenzoxazepinone BT2 that inhibits endothelial cell proliferation, migration, wound repair in vitro, network formation, and angiogenesis in mice bearing Matrigel plugs. BT2 interacts with MEK1 and inhibits ERK phosphorylation and the expression of FosB/ΔFosB, VCAM-1, and many genes involved in proliferation, migration, angiogenesis, and inflammation. BT2 reduced retinal vascular leakage following rat choroidal laser trauma and rabbit intravitreal VEGF-A165 administration. BT2 suppressed retinal CD31, pERK, VCAM-1, and VEGF-A165 expression. BT2 reduced retinal leakage in rats at least as effectively as aflibercept, a first-line therapy for nAMD/DR. BT2 withstands boiling or autoclaving and several months&#x27; storage at 22°C. BT2 is a new small-molecule inhibitor of vascular permeability and angiogenesis.},\n bibtype = {article},\n author = {Li, Y. and Alhendi, A.M.N. and Yeh, M.-C. and Elahy, M. and Santiago, F.S. and Deshpande, N.P. and Wu, B. and Chan, E. and Inam, S. and Prado-Lourenco, L. and Marcuccio, S.M. and Khachigian, L.M.},\n doi = {10.1126/sciadv.aaz7815},\n journal = {Science Advances},\n number = {31}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Vascular permeability and angiogenesis underpin neovascular age-related macular degeneration and diabetic retinopathy. While anti-VEGF therapies are widely used clinically, many patients do not respond optimally, or at all, and small-molecule therapies are lacking. Here, we identified a dibenzoxazepinone BT2 that inhibits endothelial cell proliferation, migration, wound repair in vitro, network formation, and angiogenesis in mice bearing Matrigel plugs. BT2 interacts with MEK1 and inhibits ERK phosphorylation and the expression of FosB/ΔFosB, VCAM-1, and many genes involved in proliferation, migration, angiogenesis, and inflammation. BT2 reduced retinal vascular leakage following rat choroidal laser trauma and rabbit intravitreal VEGF-A165 administration. BT2 suppressed retinal CD31, pERK, VCAM-1, and VEGF-A165 expression. BT2 reduced retinal leakage in rats at least as effectively as aflibercept, a first-line therapy for nAMD/DR. BT2 withstands boiling or autoclaving and several months' storage at 22°C. BT2 is a new small-molecule inhibitor of vascular permeability and angiogenesis.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"low-silva-sharifitabar-torrado-webb-parker-sana-smits-etal-thenucleosomeremodelinganddeacetylasecomplexhasanasymmetricdynamicandmodulararchitecture-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The Nucleosome Remodeling and Deacetylase Complex Has an Asymmetric, Dynamic, and Modular Architecture.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Low,  J.; Silva,  A.; Sharifi Tabar,  M.; Torrado,  M.; Webb,  S.; Parker,  B.; Sana,  M.; Smits,  C.; Schmidberger,  J.; Brillault,  L.; Landsberg,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Cell Reports</i>, 33(9).  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.celrep.2020.108450\"\n     onclick=\"log_download('low-silva-sharifitabar-torrado-webb-parker-sana-smits-etal-thenucleosomeremodelinganddeacetylasecomplexhasanasymmetricdynamicandmodulararchitecture-2020', 'http://doi.org/10.1016/j.celrep.2020.108450', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/low-silva-sharifitabar-torrado-webb-parker-sana-smits-etal-thenucleosomeremodelinganddeacetylasecomplexhasanasymmetricdynamicandmodulararchitecture-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('low-silva-sharifitabar-torrado-webb-parker-sana-smits-etal-thenucleosomeremodelinganddeacetylasecomplexhasanasymmetricdynamicandmodulararchitecture-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The Nucleosome Remodeling and Deacetylase Complex Has an Asymmetric, Dynamic, and Modular Architecture},\n type = {article},\n year = {2020},\n volume = {33},\n id = {a494d066-7bf4-3d91-b39e-2c81a3c17fb6},\n created = {2023-01-10T01:44:03.714Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:03.714Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Low et al. examine the architecture of the nucleosome remodeling and deacetylase complex. They define its stoichiometry, use cross-linking mass spectrometry to define subunit locations, and use electron microscopy to reveal large-scale dynamics. They also demonstrate that PWWP2A competes with MBD3 to sequester the HDAC-MTA-RBBP module from NuRD.SCOPUS_ABS_SEPARATORThe nucleosome remodeling and deacetylase (NuRD) complex is essential for metazoan development but has been refractory to biochemical analysis. We present an integrated analysis of the native mammalian NuRD complex, combining quantitative mass spectrometry, cross-linking, protein biochemistry, and electron microscopy to define the architecture of the complex. NuRD is built from a 2:2:4 (MTA, HDAC, and RBBP) deacetylase module and a 1:1:1 (MBD, GATAD2, and Chromodomain-Helicase-DNA-binding [CHD]) remodeling module, and the complex displays considerable structural dynamics. The enigmatic GATAD2 controls the asymmetry of the complex and directly recruits the CHD remodeler. The MTA-MBD interaction acts as a point of functional switching, with the transcriptional regulator PWWP2A competing with MBD for binding to the MTA-HDAC-RBBP subcomplex. Overall, our data address the long-running controversy over NuRD stoichiometry, provide imaging of the mammalian NuRD complex, and establish the biochemical mechanism by which PWWP2A can regulate NuRD composition.},\n bibtype = {article},\n author = {Low, J.K.K. and Silva, A.P.G. and Sharifi Tabar, M. and Torrado, M. and Webb, S.R. and Parker, B.L. and Sana, M. and Smits, C. and Schmidberger, J.W. and Brillault, L. and Landsberg, M.J. and Mackay, J.P.},\n doi = {10.1016/j.celrep.2020.108450},\n journal = {Cell Reports},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Low et al. examine the architecture of the nucleosome remodeling and deacetylase complex. They define its stoichiometry, use cross-linking mass spectrometry to define subunit locations, and use electron microscopy to reveal large-scale dynamics. They also demonstrate that PWWP2A competes with MBD3 to sequester the HDAC-MTA-RBBP module from NuRD.SCOPUS_ABS_SEPARATORThe nucleosome remodeling and deacetylase (NuRD) complex is essential for metazoan development but has been refractory to biochemical analysis. We present an integrated analysis of the native mammalian NuRD complex, combining quantitative mass spectrometry, cross-linking, protein biochemistry, and electron microscopy to define the architecture of the complex. NuRD is built from a 2:2:4 (MTA, HDAC, and RBBP) deacetylase module and a 1:1:1 (MBD, GATAD2, and Chromodomain-Helicase-DNA-binding [CHD]) remodeling module, and the complex displays considerable structural dynamics. The enigmatic GATAD2 controls the asymmetry of the complex and directly recruits the CHD remodeler. The MTA-MBD interaction acts as a point of functional switching, with the transcriptional regulator PWWP2A competing with MBD for binding to the MTA-HDAC-RBBP subcomplex. Overall, our data address the long-running controversy over NuRD stoichiometry, provide imaging of the mammalian NuRD complex, and establish the biochemical mechanism by which PWWP2A can regulate NuRD composition.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Peppy: A virtual reality environment for exploring the principles of polypeptide structure.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Doak,  D.; Denyer,  G.; Gerrard,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Allison,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 29(1): 157-168.  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.3752\"\n     onclick=\"log_download('doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020', 'http://doi.org/10.1002/pro.3752', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('doak-denyer-gerrard-mackay-allison-peppyavirtualrealityenvironmentforexploringtheprinciplesofpolypeptidestructure-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Peppy: A virtual reality environment for exploring the principles of polypeptide structure},\n type = {article},\n year = {2020},\n pages = {157-168},\n volume = {29},\n id = {4e51d150-054e-3635-9211-b383f4822b0e},\n created = {2023-01-10T01:44:04.638Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:04.638Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.},\n bibtype = {article},\n author = {Doak, D.G. and Denyer, G.S. and Gerrard, J.A. and Mackay, J.P. and Allison, J.R.},\n doi = {10.1002/pro.3752},\n journal = {Protein Science},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A key learning outcome for undergraduate biochemistry classes is a thorough understanding of the principles of protein structure. Traditional approaches to teaching this material, which include two-dimensional (2D) images on paper, physical molecular modeling kits, and projections of 3D structures into 2D, are unable to fully capture the dynamic 3D nature of proteins. We have built a virtual reality application, Peppy, aimed at facilitating teaching of the principles of protein secondary structure. Rather than attempt to model molecules with the same fidelity to the underlying physical chemistry as existing, research-oriented molecular modelling approaches, we took the more straightforward approach of harnessing the Unity video game physics engine. Indeed, the simplicity and limitations of our model are strengths in a teaching context, provoking questions and thus deeper understanding. Peppy allows exploration of the relative effects of hydrogen bonding (and electrostatic interactions more generally), backbone φ/ψ angles, basic chemical structure, and steric effects on a polypeptide structure in an accessible format that is novel, dynamic, and fun to use. Apart from describing the implementation and use of Peppy, we discuss the outcomes of deploying Peppy in undergraduate biochemistry courses.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"shieh-jones-vanle-au-huang-silva-lee-douine-etal-correctiongatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisordergeneticsinmedicine2020101038s414360190747z-2020\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Correction: GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder (Genetics in Medicine, (2020), 10.1038/s41436-019-0747-z).\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Shieh,  C.; Jones,  N.; Vanle,  B.; Au,  M.; Huang,  A.; Silva,  A.; Lee,  H.; Douine,  E.; Otero,  M.; Choi,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Pierson,  T.\n</span>\n\n  \n\n</span>\n\n  <i>Genetics in Medicine</i>, 22(4): 822.  2020.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41436-020-0760-2\"\n     onclick=\"log_download('shieh-jones-vanle-au-huang-silva-lee-douine-etal-correctiongatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisordergeneticsinmedicine2020101038s414360190747z-2020', 'http://doi.org/10.1038/s41436-020-0760-2', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/shieh-jones-vanle-au-huang-silva-lee-douine-etal-correctiongatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisordergeneticsinmedicine2020101038s414360190747z-2020\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('shieh-jones-vanle-au-huang-silva-lee-douine-etal-correctiongatad2bassociatedneurodevelopmentaldisordergandclinicalandmolecularinsightsintoanurdrelateddisordergeneticsinmedicine2020101038s414360190747z-2020')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Correction: GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder (Genetics in Medicine, (2020), 10.1038/s41436-019-0747-z)},\n type = {article},\n year = {2020},\n pages = {822},\n volume = {22},\n id = {7ee81c43-2642-362b-af4d-d6af97c63161},\n created = {2023-01-10T01:44:05.579Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:05.579Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {An amendment to this paper has been published and can be accessed via a link at the top of the paper.},\n bibtype = {article},\n author = {Shieh, C. and Jones, N. and Vanle, B. and Au, M. and Huang, A.Y. and Silva, A.P.G. and Lee, H. and Douine, E.D. and Otero, M.G. and Choi, A. and Mackay, J.P. and Pierson, T.M.},\n doi = {10.1038/s41436-020-0760-2},\n journal = {Genetics in Medicine},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  An amendment to this paper has been published and can be accessed via a link at the top of the paper.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2019\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2019')\"\n      onkeypress=\"toggleGroup('group_2019')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2019</span>\n      <span class=\"bibbase_group_count\">\n        \n        (6)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.org/10.1002/ajmg.c.31752\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019', 'http://doi.org/10.1002/ajmg.c.31752', event);\">\n    The NuRD complex and macrocephaly associated neurodevelopmental disorders.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Pierson,  T., M.; Otero,  M., G.; Grand,  K.; Choi,  A.; Graham,  J., M.; Young,  J., I.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>American Journal of Medical Genetics Part C: Seminars in Medical Genetics</i>, 181(4): 548-556. 12 2019.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.org/10.1002/ajmg.c.31752\"\n     onclick=\"log_download('pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019', 'http://doi.org/10.1002/ajmg.c.31752', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"The NuRD complex and macrocephaly associated neurodevelopmental disorders [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/ajmg.c.31752\"\n     onclick=\"log_download('pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019', 'http://doi.org/10.1002/ajmg.c.31752', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The NuRD complex and macrocephaly associated neurodevelopmental disorders},\n type = {article},\n year = {2019},\n keywords = {CHD3,CHD4,GATAD2B,NuRD complex,macrocephaly},\n pages = {548-556},\n volume = {181},\n websites = {http://doi.org/10.1002/ajmg.c.31752},\n month = {12},\n day = {18},\n id = {786626bb-912b-3ca7-aa02-82a98a6afa89},\n created = {2020-12-17T05:29:52.824Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:52.824Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Pierson2019},\n source_type = {article},\n private_publication = {false},\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.},\n bibtype = {article},\n author = {Pierson, Tyler Mark and Otero, Maria G. and Grand, Katheryn and Choi, Andrew and Graham, John M. and Young, Juan I. and Mackay, Joel P.},\n doi = {10.1002/ajmg.c.31752},\n journal = {American Journal of Medical Genetics Part C: Seminars in Medical Genetics},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.org/10.1002/prot.25687\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019', 'http://doi.org/10.1002/prot.25687', event);\">\n    The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mohanty,  B.; Hanson‐Manful,  P.; Finn,  T., J.; Chambers,  C., R.; McKellar,  J., L., O.; Macindoe,  I.; Helder,  S.; Setiyaputra,  S.; Zhong,  Y.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Patrick,  W., M.\n</span>\n\n  \n\n</span>\n\n  <i>Proteins: Structure, Function, and Bioinformatics</i>, 87(8): 699-705. 8 2019.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.org/10.1002/prot.25687\"\n     onclick=\"log_download('mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019', 'http://doi.org/10.1002/prot.25687', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/prot.25687\"\n     onclick=\"log_download('mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019', 'http://doi.org/10.1002/prot.25687', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G},\n type = {article},\n year = {2019},\n keywords = {IPR020489,NMR,PF13989,RNA recognition motif,aminoglycoside,elongation factor},\n pages = {699-705},\n volume = {87},\n websites = {http://doi.org/10.1002/prot.25687},\n month = {8},\n day = {22},\n id = {7fb760d5-9bf0-3ae6-b357-d8046463887d},\n created = {2020-12-17T05:29:57.892Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:57.892Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Mohanty2019},\n source_type = {article},\n private_publication = {false},\n abstract = {InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.},\n bibtype = {article},\n author = {Mohanty, Biswaranjan and Hanson‐Manful, Paulina and Finn, Thomas J. and Chambers, Cecilia R. and McKellar, James L. O. and Macindoe, Ingrid and Helder, Stephanie and Setiyaputra, Surya and Zhong, Yichen and Mackay, Joel P. and Patrick, Wayne M.},\n doi = {10.1002/prot.25687},\n journal = {Proteins: Structure, Function, and Bioinformatics},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mohanty,  B.; Hanson-Manful,  P.; Finn,  T.; Chambers,  C.; McKellar,  J.; Macindoe,  I.; Helder,  S.; Setiyaputra,  S.; Zhong,  Y.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Patrick,  W.\n</span>\n\n  \n\n</span>\n\n  <i>Proteins: Structure, Function and Bioinformatics</i>, 87(8): 699-705.  2019.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/prot.25687\"\n     onclick=\"log_download('mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019', 'http://doi.org/10.1002/prot.25687', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mohanty-hansonmanful-finn-chambers-mckellar-macindoe-helder-setiyaputra-etal-theuncharacterizedbacterialproteinyejghasthesamearchitectureasdomainiiiofelongationfactorg-2019')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The uncharacterized bacterial protein YejG has the same architecture as domain III of elongation factor G},\n type = {article},\n year = {2019},\n pages = {699-705},\n volume = {87},\n id = {eef77526-79b2-36df-bdf3-98887024df8f},\n created = {2023-01-10T01:44:06.506Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:06.506Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.},\n bibtype = {article},\n author = {Mohanty, B. and Hanson-Manful, P. and Finn, T.J. and Chambers, C.R. and McKellar, J.L.O. and Macindoe, I. and Helder, S. and Setiyaputra, S. and Zhong, Y. and Mackay, J.P. and Mackay, J.P. and Patrick, W.M.},\n doi = {10.1002/prot.25687},\n journal = {Proteins: Structure, Function and Bioinformatics},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  InterPro family IPR020489 comprises ~1000 uncharacterized bacterial proteins. Previously we showed that overexpressing the Escherichia coli representative of this family, EcYejG, conferred low-level resistance to aminoglycoside antibiotics. In an attempt to shed light on the biochemical function of EcYejG, we have solved its structure using multinuclear solution NMR spectroscopy. The structure most closely resembles that of domain III from elongation factor G (EF-G). EF-G catalyzes ribosomal translocation and mutations in EF-G have also been associated with aminoglycoside resistance. While we were unable to demonstrate a direct interaction between EcYejG and the ribosome, the protein might play a role in translation.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"sharifitabar-mackay-low-thestoichiometryandinteractomeofthenucleosomeremodelinganddeacetylasenurdcomplexareconservedacrossmultiplecelllines-2019\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The stoichiometry and interactome of the Nucleosome Remodeling and Deacetylase (NuRD) complex are conserved across multiple cell lines.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Sharifi Tabar,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Low,  J.\n</span>\n\n  \n\n</span>\n\n  <i>FEBS Journal</i>, 286(11): 2043-2061.  2019.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1111/febs.14800\"\n     onclick=\"log_download('sharifitabar-mackay-low-thestoichiometryandinteractomeofthenucleosomeremodelinganddeacetylasenurdcomplexareconservedacrossmultiplecelllines-2019', 'http://doi.org/10.1111/febs.14800', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/sharifitabar-mackay-low-thestoichiometryandinteractomeofthenucleosomeremodelinganddeacetylasenurdcomplexareconservedacrossmultiplecelllines-2019\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>3 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('sharifitabar-mackay-low-thestoichiometryandinteractomeofthenucleosomeremodelinganddeacetylasenurdcomplexareconservedacrossmultiplecelllines-2019')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The stoichiometry and interactome of the Nucleosome Remodeling and Deacetylase (NuRD) complex are conserved across multiple cell lines},\n type = {article},\n year = {2019},\n pages = {2043-2061},\n volume = {286},\n id = {51b93072-bfe2-37ed-9321-35234e2dd662},\n created = {2023-01-10T01:44:07.415Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:07.415Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The nucleosome remodelling and deacetylase complex (NuRD) is a widely conserved regulator of gene expression. The determination of the subunit composition of the complex and identification of its binding partners are important steps towards understanding its architecture and function. The question of how these properties of the complex vary across different cell types has not been addressed in detail to date. Here, we set up a two-step purification protocol coupled to liquid chromatography-tandem mass spectrometry to assess NuRD composition and interaction partners in three different cancer cell lines, using label-free intensity-based absolute quantification (iBAQ). Our data indicate that the stoichiometry of the NuRD complex is preserved across our three different cancer cell lines. In addition, our interactome data suggest ZNF219 and SLC25A5 as possible interaction partners of the complex. To corroborate this latter finding, in vitro and cell-based pull-down experiments were carried out. These experiments indicated that ZNF219 can interact with RBBP4, GATAD2A/B and chromodomain helicase DNA binding 4, whereas SLC25A5 might interact with MTA2 and GATAD2A.},\n bibtype = {article},\n author = {Sharifi Tabar, M. and Mackay, J.P. and Low, J.K.K.},\n doi = {10.1111/febs.14800},\n journal = {FEBS Journal},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nucleosome remodelling and deacetylase complex (NuRD) is a widely conserved regulator of gene expression. The determination of the subunit composition of the complex and identification of its binding partners are important steps towards understanding its architecture and function. The question of how these properties of the complex vary across different cell types has not been addressed in detail to date. Here, we set up a two-step purification protocol coupled to liquid chromatography-tandem mass spectrometry to assess NuRD composition and interaction partners in three different cancer cell lines, using label-free intensity-based absolute quantification (iBAQ). Our data indicate that the stoichiometry of the NuRD complex is preserved across our three different cancer cell lines. In addition, our interactome data suggest ZNF219 and SLC25A5 as possible interaction partners of the complex. To corroborate this latter finding, in vitro and cell-based pull-down experiments were carried out. These experiments indicated that ZNF219 can interact with RBBP4, GATAD2A/B and chromodomain helicase DNA binding 4, whereas SLC25A5 might interact with MTA2 and GATAD2A.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"defranco-vandenameele-brans-verlaine-bendak-damblon-matagne-segal-etal-exploringthesuitabilityofranbp2typezincfingersforrnabindingproteindesign-2019\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Exploring the suitability of RanBP2-type Zinc Fingers for RNA-binding protein design.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  De Franco,  S.; Vandenameele,  J.; Brans,  A.; Verlaine,  O.; Bendak,  K.; Damblon,  C.; Matagne,  A.; Segal,  D.; Galleni,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Vandevenne,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Scientific Reports</i>, 9(1).  2019.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41598-019-38655-y\"\n     onclick=\"log_download('defranco-vandenameele-brans-verlaine-bendak-damblon-matagne-segal-etal-exploringthesuitabilityofranbp2typezincfingersforrnabindingproteindesign-2019', 'http://doi.org/10.1038/s41598-019-38655-y', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/defranco-vandenameele-brans-verlaine-bendak-damblon-matagne-segal-etal-exploringthesuitabilityofranbp2typezincfingersforrnabindingproteindesign-2019\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>6 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('defranco-vandenameele-brans-verlaine-bendak-damblon-matagne-segal-etal-exploringthesuitabilityofranbp2typezincfingersforrnabindingproteindesign-2019')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Exploring the suitability of RanBP2-type Zinc Fingers for RNA-binding protein design},\n type = {article},\n year = {2019},\n volume = {9},\n id = {221c72ab-8395-34b9-89e2-2cc6f2d798f7},\n created = {2023-01-10T01:44:08.343Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:08.343Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Transcriptomes consist of several classes of RNA that have wide-ranging but often poorly described functions and the deregulation of which leads to numerous diseases. Engineering of functionalized RNA-binding proteins (RBPs) could therefore have many applications. Our previous studies suggested that the RanBP2-type Zinc Finger (ZF) domain is a suitable scaffold to investigate the design of single-stranded RBPs. In the present work, we have analyzed the natural sequence specificity of various members of the RanBP2-type ZF family and characterized the interaction with their target RNA. Surprisingly, our data showed that natural RanBP2-type ZFs with different RNA-binding residues exhibit a similar sequence specificity and therefore no simple recognition code can be established. Despite this finding, different discriminative abilities were observed within the family. In addition, in order to target a long RNA sequence and therefore gain in specificity, we generated a 6-ZF array by combining ZFs from the RanBP2-type family but also from different families, in an effort to achieve a wider target sequence repertoire. We showed that this chimeric protein recognizes its target sequence (20 nucleotides), both in vitro and in living cells. Altogether, our results indicate that the use of ZFs in RBP design remains attractive even though engineering of specificity changes is challenging.},\n bibtype = {article},\n author = {De Franco, S. and Vandenameele, J. and Brans, A. and Verlaine, O. and Bendak, K. and Damblon, C. and Matagne, A. and Segal, D.J. and Galleni, M. and Mackay, J.P. and Mackay, J.P. and Vandevenne, M.},\n doi = {10.1038/s41598-019-38655-y},\n journal = {Scientific Reports},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Transcriptomes consist of several classes of RNA that have wide-ranging but often poorly described functions and the deregulation of which leads to numerous diseases. Engineering of functionalized RNA-binding proteins (RBPs) could therefore have many applications. Our previous studies suggested that the RanBP2-type Zinc Finger (ZF) domain is a suitable scaffold to investigate the design of single-stranded RBPs. In the present work, we have analyzed the natural sequence specificity of various members of the RanBP2-type ZF family and characterized the interaction with their target RNA. Surprisingly, our data showed that natural RanBP2-type ZFs with different RNA-binding residues exhibit a similar sequence specificity and therefore no simple recognition code can be established. Despite this finding, different discriminative abilities were observed within the family. In addition, in order to target a long RNA sequence and therefore gain in specificity, we generated a 6-ZF array by combining ZFs from the RanBP2-type family but also from different families, in an effort to achieve a wider target sequence repertoire. We showed that this chimeric protein recognizes its target sequence (20 nucleotides), both in vitro and in living cells. Altogether, our results indicate that the use of ZFs in RBP design remains attractive even though engineering of specificity changes is challenging.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The NuRD complex and macrocephaly associated neurodevelopmental disorders.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Pierson,  T.; Otero,  M.; Grand,  K.; Choi,  A.; Graham,  J.; Young,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>American Journal of Medical Genetics, Part C: Seminars in Medical Genetics</i>, 181(4): 548-556.  2019.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/ajmg.c.31752\"\n     onclick=\"log_download('pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019', 'http://doi.org/10.1002/ajmg.c.31752', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('pierson-otero-grand-choi-graham-young-mackay-thenurdcomplexandmacrocephalyassociatedneurodevelopmentaldisorders-2019')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The NuRD complex and macrocephaly associated neurodevelopmental disorders},\n type = {article},\n year = {2019},\n pages = {548-556},\n volume = {181},\n id = {03edfe18-463d-3f00-9f0b-586a1515ccc8},\n created = {2023-01-10T01:44:09.267Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:09.267Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.},\n bibtype = {article},\n author = {Pierson, T.M. and Otero, M.G. and Grand, K. and Choi, A. and Graham, J.M. and Young, J.I. and Mackay, J.P.},\n doi = {10.1002/ajmg.c.31752},\n journal = {American Journal of Medical Genetics, Part C: Seminars in Medical Genetics},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nucleosome remodeling and deacetylase (NuRD) complex is a major regulator of gene expression involved in pluripotency, lineage commitment, and corticogenesis. This important complex is composed of seven different proteins, with mutations in CHD3, CHD4, and GATAD2B being associated with neurodevelopmental disorders presenting with macrocephaly and intellectual disability similar to other overgrowth and intellectual disability (OGID) syndromes. Pathogenic variants in CHD3 and CHD4 primarily involve disruption of enzymatic function. GATAD2B variants include loss-of-function mutations that alter protein dosage and missense variants that involve either of two conserved domains (CR1 and CR2) known to interact with other NuRD proteins. In addition to macrocephaly and intellectual disability, CHD3 variants are associated with inguinal hernias and apraxia of speech; whereas CHD4 variants are associated with skeletal anomalies, deafness, and cardiac defects. GATAD2B-associated neurodevelopmental disorder (GAND) has phenotypic overlap with both of these disorders. Of note, structural models of NuRD indicate that CHD3 and CHD4 require direct contact with the GATAD2B-CR2 domain to interact with the rest of the complex. Therefore, the phenotypic overlaps of CHD3- and CHD4-related disorders with GAND are consistent with a loss in the ability of GATAD2B to recruit CHD3 or CHD4 to the complex. The shared features of these neurodevelopmental disorders may represent a new class of OGID syndrome: the NuRDopathies.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2018\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2018')\"\n      onkeypress=\"toggleGroup('group_2018')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2018</span>\n      <span class=\"bibbase_group_count\">\n        \n        (11)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1111/mpp.12597\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018', 'http://doi.wiley.com/10.1111/mpp.12597', event);\">\n    Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Zhang,  X.; Farah,  N.; Rolston,  L.; Ericsson,  D., J.; Catanzariti,  A.; Bernoux,  M.; Ve,  T.; Bendak,  K.; Chen,  C.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Lawrence,  G., J.; Hardham,  A.; Ellis,  J., G.; Williams,  S., J.; Dodds,  P., N.; Jones,  D., A.;  and Kobe,  B.\n</span>\n\n  \n\n</span>\n\n  <i>Molecular Plant Pathology</i>, 19(5): 1196-1209. 5 2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1111/mpp.12597\"\n     onclick=\"log_download('zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018', 'http://doi.wiley.com/10.1111/mpp.12597', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1111/mpp.12597\"\n     onclick=\"log_download('zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018', 'http://doi.org/10.1111/mpp.12597', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P},\n type = {article},\n year = {2018},\n keywords = {NLR [nucleotide-binding and oligomerization domain,crystal structure,effector-triggered immunity,flax rust (Melampsora lini) effector,nuclear localization,nucleotide-binding/leucine-rich repeat receptor],plant disease resistance,zinc finger},\n pages = {1196-1209},\n volume = {19},\n websites = {http://doi.wiley.com/10.1111/mpp.12597},\n month = {5},\n id = {67c03f3e-7a41-34e7-89cf-55716f13ac32},\n created = {2020-12-17T05:29:53.248Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:53.248Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Zhang2018},\n source_type = {ARTICLE},\n notes = {cited By 2},\n private_publication = {false},\n abstract = {The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.},\n bibtype = {article},\n author = {Zhang, Xiaoxiao and Farah, Nadya and Rolston, Laura and Ericsson, Daniel J. and Catanzariti, Ann-Maree and Bernoux, Maud and Ve, Thomas and Bendak, Katerina and Chen, Chunhong and Mackay, Joel P. and Lawrence, Gregory J. and Hardham, Adrienne and Ellis, Jeffrey G. and Williams, Simon J. and Dodds, Peter N. and Jones, David A. and Kobe, Bostjan},\n doi = {10.1111/mpp.12597},\n journal = {Molecular Plant Pathology},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"moghaddassani-hamzehmivehroud-silva-walshe-mohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://bi.tbzmed.ac.ir/Abstract/bi-17551\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'moghaddassani-hamzehmivehroud-silva-walshe-mohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018', 'http://bi.tbzmed.ac.ir/Abstract/bi-17551', event);\">\n    Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Moghaddas Sani,  H.; Hamzeh-Mivehroud,  M.; Silva,  A., P.; Walshe,  J., L.; Mohammadi,  S., A.; Rahbar-Shahrouziasl,  M.; Abbasi,  M.; Jamshidi,  O.; Low,  J., K.; Dastmalchi,  S.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>BioImpacts</i>, 8(3): 167-176. 1 2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://bi.tbzmed.ac.ir/Abstract/bi-17551\"\n     onclick=\"log_download('moghaddassani-hamzehmivehroud-silva-walshe-mohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018', 'http://bi.tbzmed.ac.ir/Abstract/bi-17551', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.15171/bi.2018.19\"\n     onclick=\"log_download('moghaddassani-hamzehmivehroud-silva-walshe-mohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018', 'http://doi.org/10.15171/bi.2018.19', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/moghaddassani-hamzehmivehroud-silva-walshe-mohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('moghaddassani-hamzehmivehroud-silva-walshe-mohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana},\n type = {article},\n year = {2018},\n keywords = {DNA binding affinity,DOF zinc finger domain,Gel retardation assay,Microscale thermophoresis},\n pages = {167-176},\n volume = {8},\n websites = {http://bi.tbzmed.ac.ir/Abstract/bi-17551},\n month = {1},\n day = {28},\n id = {dd4675dd-58e2-3545-8030-ebe4c5b425a8},\n created = {2020-12-17T05:29:55.192Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:55.192Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {MoghaddasSani2018},\n source_type = {ARTICLE},\n notes = {cited By 1},\n private_publication = {false},\n abstract = {Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.},\n bibtype = {article},\n author = {Moghaddas Sani, Hakimeh and Hamzeh-Mivehroud, Maryam and Silva, Ana P. and Walshe, James L. and Mohammadi, S. Abolghasem and Rahbar-Shahrouziasl, Mahdyieh and Abbasi, Milad and Jamshidi, Omid and Low, Jason K.K. and Dastmalchi, Siavoush and Mackay, Joel P.},\n doi = {10.15171/bi.2018.19},\n journal = {BioImpacts},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1111/jth.13900\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018', 'http://doi.wiley.com/10.1111/jth.13900', event);\">\n    Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mundell,  S., J.; Rabbolini,  D.; Gabrielli,  S.; Chen,  Q.; Aungraheeta,  R.; Hutchinson,  J., L.; Kilo,  T.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Ward,  C., M.; Stevenson,  W.;  and Morel-Kopp,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Thrombosis and Haemostasis</i>, 16(1): 44-53. 1 2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1111/jth.13900\"\n     onclick=\"log_download('mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018', 'http://doi.wiley.com/10.1111/jth.13900', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1111/jth.13900\"\n     onclick=\"log_download('mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018', 'http://doi.org/10.1111/jth.13900', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding},\n type = {article},\n year = {2018},\n keywords = {P2RY12,blood platelet disorder,human,inherited,platelet dysfunction},\n pages = {44-53},\n volume = {16},\n websites = {http://doi.wiley.com/10.1111/jth.13900},\n month = {1},\n id = {6ba6e7ee-21d0-3efa-a162-8e61a57a2e8e},\n created = {2020-12-17T05:29:57.411Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:57.411Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Mundell2018},\n source_type = {ARTICLE},\n notes = {cited By 1},\n private_publication = {false},\n abstract = {Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.},\n bibtype = {article},\n author = {Mundell, S. J. and Rabbolini, D. and Gabrielli, S. and Chen, Q. and Aungraheeta, R. and Hutchinson, J. L. and Kilo, T. and Mackay, J. and Ward, C. M. and Stevenson, W. and Morel-Kopp, M.-C.},\n doi = {10.1111/jth.13900},\n journal = {Journal of Thrombosis and Haemostasis},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gell-kwan-mackay-modernmagneticresonance-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://link.springer.com/10.1007/978-3-319-28388-3_121\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'gell-kwan-mackay-modernmagneticresonance-2018', 'http://link.springer.com/10.1007/978-3-319-28388-3_121', event);\">\n    NMR Spectroscopy in the Analysis of Protein-Protein Interactions.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gell,  D., A.; Kwan,  A., H.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  Modern Magnetic Resonance, pages 2099-2132. Springer International Publishing,  2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://link.springer.com/10.1007/978-3-319-28388-3_121\"\n     onclick=\"log_download('gell-kwan-mackay-modernmagneticresonance-2018', 'http://link.springer.com/10.1007/978-3-319-28388-3_121', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Modern Magnetic Resonance [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1007/978-3-319-28388-3_121\"\n     onclick=\"log_download('gell-kwan-mackay-modernmagneticresonance-2018', 'http://doi.org/10.1007/978-3-319-28388-3_121', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gell-kwan-mackay-modernmagneticresonance-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gell-kwan-mackay-modernmagneticresonance-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@inbook{\n type = {inbook},\n year = {2018},\n keywords = {Chemical exchange,Chemical shift perturbation,Cross-saturation,Dark states,Macromolecular NMR spectroscopy,Methyl-TROSY,Protein complexes,Protein-protein interactions},\n pages = {2099-2132},\n websites = {http://link.springer.com/10.1007/978-3-319-28388-3_121},\n publisher = {Springer International Publishing},\n city = {Cham},\n id = {49e6d065-5bfa-3734-ac4c-302065de5fd0},\n created = {2020-12-17T05:29:58.281Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:58.281Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Gell2018},\n source_type = {article},\n private_publication = {false},\n abstract = {Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.},\n bibtype = {inbook},\n author = {Gell, David A. and Kwan, Ann H. and Mackay, Joel P.},\n doi = {10.1007/978-3-319-28388-3_121},\n chapter = {NMR Spectroscopy in the Analysis of Protein-Protein Interactions},\n title = {Modern Magnetic Resonance}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mundell,  S.; Rabbolini,  D.; Gabrielli,  S.; Chen,  Q.; Aungraheeta,  R.; Hutchinson,  J.; Kilo,  T.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Ward,  C.; Stevenson,  W.; Stevenson,  W.;  and Morel-Kopp,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Thrombosis and Haemostasis</i>, 16(1): 44-53.  2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1111/jth.13900\"\n     onclick=\"log_download('mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018', 'http://doi.org/10.1111/jth.13900', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mundell-rabbolini-gabrielli-chen-aungraheeta-hutchinson-kilo-mackay-etal-receptorhomodimerizationplaysacriticalroleinanoveldominantnegativep2ry12variantidentifiedinafamilywithseverebleeding-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Receptor homodimerization plays a critical role in a novel dominant negative P2RY12 variant identified in a family with severe bleeding},\n type = {article},\n year = {2018},\n pages = {44-53},\n volume = {16},\n id = {086665b6-9995-388c-8525-8bf8e6dfb235},\n created = {2023-01-10T01:44:10.193Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:10.193Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.},\n bibtype = {article},\n author = {Mundell, S.J. and Rabbolini, D. and Gabrielli, S. and Chen, Q. and Aungraheeta, R. and Hutchinson, J.L. and Kilo, T. and Mackay, J. and Ward, C.M. and Stevenson, W. and Stevenson, W. and Morel-Kopp, M.-C.},\n doi = {10.1111/jth.13900},\n journal = {Journal of Thrombosis and Haemostasis},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Essentials Three dominant variants for the autosomal recessive bleeding disorder type-8 have been described. To date, there has been no phenotype/genotype correlation explaining their dominant transmission. Proline plays an important role in P2Y12R ligand binding and signaling defects. P2Y12R homodimer formation is critical for the receptor function and signaling. Summary: Background Although inherited platelet disorders are still underdiagnosed worldwide, advances in molecular techniques are improving disease diagnosis and patient management. Objective To identify and characterize the mechanism underlying the bleeding phenotype in a Caucasian family with an autosomal dominant P2RY12 variant. Methods Full blood counts, platelet aggregometry, flow cytometry and western blotting were performed before next-generation sequencing (NGS). Detailed molecular analysis of the identified variant of the P2Y12 receptor (P2Y12R) was subsequently performed in mammalian cells overexpressing receptor constructs. Results All three referred individuals had markedly impaired ADP-induced platelet aggregation with primary wave only, despite normal total and surface P2Y12R expression. By NGS, a single P2RY12:c.G794C substitution (p.R265P) was identified in all affected individuals, and this was confirmed by Sanger sequencing. Mammalian cell experiments with the R265P-P2Y12R variant showed normal receptor surface expression versus wild-type (WT) P2Y12R. Agonist-stimulated R265P-P2Y12R function (both signaling and surface receptor loss) was reduced versus WT P2Y12R. Critically, R265P-P2Y12R acted in a dominant negative manner, with agonist-stimulated WT P2Y12R activity being reduced by variant coexpression, suggesting dramatic loss of WT homodimers. Importantly, platelet P2RY12 cDNA cloning and sequencing in two affected individuals also revealed three-fold mutant mRNA overexpression, decreasing even further the likelihood of WT homodimer formation. R265 located within extracellular loop 3 (EL3) is one of four residues that are important for receptor functional integrity, maintaining the binding pocket conformation and allowing rotation following ligand binding. Conclusion This novel dominant negative variant confirms the important role of R265 in EL3 in the functional integrity of P2Y12R, and suggests that pathologic heterodimer formation may underlie this family bleeding phenotype.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"sani-hamzehmivehroud-silva-walshe-abolghasemmohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Sani,  H.; Hamzeh-Mivehroud,  M.; Silva,  A.; Walshe,  J.; Abolghasem Mohammadi,  S.; Rahbar-Shahrouziasl,  M.; Abbasi,  M.; Jamshidi,  O.; Low,  J.; Dastmalchi,  S.; Dastmalchi,  S.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>BioImpacts</i>, 8(3): 167-176.  2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.15171/bi.2018.19\"\n     onclick=\"log_download('sani-hamzehmivehroud-silva-walshe-abolghasemmohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018', 'http://doi.org/10.15171/bi.2018.19', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/sani-hamzehmivehroud-silva-walshe-abolghasemmohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('sani-hamzehmivehroud-silva-walshe-abolghasemmohammadi-rahbarshahrouziasl-abbasi-jamshidi-etal-expressionpurificationanddnabindingpropertiesofzincfingerdomainsofdofproteinsfromarabidopsisthaliana-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Expression, purification and DNA-binding properties of zinc finger domains of DOF proteins from Arabidopsis thaliana},\n type = {article},\n year = {2018},\n pages = {167-176},\n volume = {8},\n id = {dd057f52-1171-342a-ba86-9f93ba6db989},\n created = {2023-01-10T01:44:11.110Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:11.110Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.},\n bibtype = {article},\n author = {Sani, H.M. and Hamzeh-Mivehroud, M. and Silva, A.P. and Walshe, J.L. and Abolghasem Mohammadi, S. and Rahbar-Shahrouziasl, M. and Abbasi, M. and Jamshidi, O. and Low, J.K.K. and Dastmalchi, S. and Dastmalchi, S. and Mackay, J.P.},\n doi = {10.15171/bi.2018.19},\n journal = {BioImpacts},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Introduction: DOF proteins are a family of plant-specific transcription factors with a conserved zinc finger (ZF) DNA-binding domain. Although several studies have demonstrated their specific DNA binding, quantitative affinity data is not available for the binding of DOF domains to their binding sites. Methods: ZF domains of DOF2.1, DOF3.4, and DOF5.8 from Arabidopsis thaliana were expressed and purified. Their DNA binding affinities were assessed using gel retardation assays and microscale thermophoresis with two different oligonucleotide probes containing one and two copies of recognition sequence AAAG. Results: DOF zinc finger domains (DOF-ZFs) were shown to form independently folded structures. Assessments using microscale thermophoresis demonstrated that DOF-ZFs interact more tightly (~100 fold) with double-motif probe than the single-motif probe. The overall Kd values for the DOF3.4-ZF and DOF5.8-ZF to the double-motif probe were ~2.3±1 and 2.5±1 μM, respectively. Conclusion: Studied DOF-ZF domains formed stable complexes with the double-motif probe. Although DOF3.4-ZF and DOF5.8-ZF do not dimerize with an appreciable affinity in the absence of DNA (judging from size-exclusion and multiangle laser light scattering data), it is possible that these ZFs form protein-protein contacts when bound to this oligonucleotide, consistent with previous reports that DOF proteins can homo- and hetero-dimerize.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"link-spitzer-sana-torrado-vlkeralbert-keilhauer-burgold-pnzeler-etal-pwwp2abindsdistinctchromatinmoietiesandinteractswithanmta1specificcorenurdcomplex-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  PWWP2A binds distinct chromatin moieties and interacts with an MTA1-specific core NuRD complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Link,  S.; Spitzer,  R.; Sana,  M.; Torrado,  M.; Völker-Albert,  M.; Keilhauer,  E.; Burgold,  T.; Pünzeler,  S.; Low,  J.; Lindström,  I.; Bartkuhn,  M.;  and Hake,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Communications</i>, 9(1).  2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/s41467-018-06665-5\"\n     onclick=\"log_download('link-spitzer-sana-torrado-vlkeralbert-keilhauer-burgold-pnzeler-etal-pwwp2abindsdistinctchromatinmoietiesandinteractswithanmta1specificcorenurdcomplex-2018', 'http://doi.org/10.1038/s41467-018-06665-5', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/link-spitzer-sana-torrado-vlkeralbert-keilhauer-burgold-pnzeler-etal-pwwp2abindsdistinctchromatinmoietiesandinteractswithanmta1specificcorenurdcomplex-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('link-spitzer-sana-torrado-vlkeralbert-keilhauer-burgold-pnzeler-etal-pwwp2abindsdistinctchromatinmoietiesandinteractswithanmta1specificcorenurdcomplex-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {PWWP2A binds distinct chromatin moieties and interacts with an MTA1-specific core NuRD complex},\n type = {article},\n year = {2018},\n volume = {9},\n id = {3ac8e87c-41a6-3ef1-859c-d226ac51ccaa},\n created = {2023-01-10T01:44:12.035Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:12.035Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Chromatin structure and function is regulated by reader proteins recognizing histone modifications and/or histone variants. We recently identified that PWWP2A tightly binds to H2A.Z-containing nucleosomes and is involved in mitotic progression and cranial–facial development. Here, using in vitro assays, we show that distinct domains of PWWP2A mediate binding to free linker DNA as well as H3K36me3 nucleosomes. In vivo, PWWP2A strongly recognizes H2A.Z-containing regulatory regions and weakly binds H3K36me3-containing gene bodies. Further, PWWP2A binds to an MTA1-specific subcomplex of the NuRD complex (M1HR), which consists solely of MTA1, HDAC1, and RBBP4/7, and excludes CHD, GATAD2 and MBD proteins. Depletion of PWWP2A leads to an increase of acetylation levels on H3K27 as well as H2A.Z, presumably by impaired chromatin recruitment of M1HR. Thus, this study identifies PWWP2A as a complex chromatin-binding protein that serves to direct the deacetylase complex M1HR to H2A.Z-containing chromatin, thereby promoting changes in histone acetylation levels.},\n bibtype = {article},\n author = {Link, S. and Spitzer, R.M.M. and Sana, M. and Torrado, M. and Völker-Albert, M.C. and Keilhauer, E.C. and Burgold, T. and Pünzeler, S. and Low, J.K.K. and Lindström, I. and Bartkuhn, M. and Hake, S.B.},\n doi = {10.1038/s41467-018-06665-5},\n journal = {Nature Communications},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chromatin structure and function is regulated by reader proteins recognizing histone modifications and/or histone variants. We recently identified that PWWP2A tightly binds to H2A.Z-containing nucleosomes and is involved in mitotic progression and cranial–facial development. Here, using in vitro assays, we show that distinct domains of PWWP2A mediate binding to free linker DNA as well as H3K36me3 nucleosomes. In vivo, PWWP2A strongly recognizes H2A.Z-containing regulatory regions and weakly binds H3K36me3-containing gene bodies. Further, PWWP2A binds to an MTA1-specific subcomplex of the NuRD complex (M1HR), which consists solely of MTA1, HDAC1, and RBBP4/7, and excludes CHD, GATAD2 and MBD proteins. Depletion of PWWP2A leads to an increase of acetylation levels on H3K27 as well as H2A.Z, presumably by impaired chromatin recruitment of M1HR. Thus, this study identifies PWWP2A as a complex chromatin-binding protein that serves to direct the deacetylase complex M1HR to H2A.Z-containing chromatin, thereby promoting changes in histone acetylation levels.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gell-kwan-mackay-nmrspectroscopyintheanalysisofproteinproteininteractions-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  NMR spectroscopy in the analysis of protein-protein interactions.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gell,  D.; Kwan,  A.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n   2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/978-3-319-28388-3_121\"\n     onclick=\"log_download('gell-kwan-mackay-nmrspectroscopyintheanalysisofproteinproteininteractions-2018', 'http://doi.org/10.1007/978-3-319-28388-3_121', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gell-kwan-mackay-nmrspectroscopyintheanalysisofproteinproteininteractions-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gell-kwan-mackay-nmrspectroscopyintheanalysisofproteinproteininteractions-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@book{\n title = {NMR spectroscopy in the analysis of protein-protein interactions},\n type = {book},\n year = {2018},\n source = {Modern Magnetic Resonance},\n pages = {2099-2132},\n id = {8844e241-63ec-3ea2-bdbe-1d62e7545c91},\n created = {2023-01-10T01:44:12.957Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:12.957Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.},\n bibtype = {book},\n author = {Gell, D.A. and Kwan, A.H. and Mackay, J.P.},\n doi = {10.1007/978-3-319-28388-3_121}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Protein-protein interactions are a central aspect of biology and NMR spectroscopy is one of the most powerful and versatile methods available to characterize their structure, dynamics, kinetics and thermodynamics. In this article, we give an overview of the suite of approaches available to the researcher who wishes to understand their favourite protein-protein interaction in more detail. We begin with an outline of two fundamental concepts that are important for understanding the strengths and limitations of NMR spectroscopy - nuclear spin relaxation and chemical exchange. We then present a range of methods including chemical shift perturbation analysis, nuclear Overhauser effects (and its derivatives), residual dipolar couplings, paramagnetic approaches, solid-state NMR and the analysis of low-abundance species. Each method is accompanied by recen texamples from the literature. Together, these techniques can allow both broad and deep insight into the mechanistic underpinnings of protein-protein interactions.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wai-szyszka-campbell-kwong-lorna-silva-low-kwan-etal-thebrd3etdomainrecognizesashortpeptidemotifthroughamechanismthatisconservedacrosschromatinremodelersandtranscriptionalregulators-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The BRD3 ET domain recognizes a short peptide motif through a mechanism that is conserved across chromatin remodelers and transcriptional regulators.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wai,  D.; Szyszka,  T.; Campbell,  A.; Kwong,  C.; Lorna,  E.; Silva,  A.; Low,  J.; Kwan,  A.; Gamsjaeger,  R.; Chalmers,  J.; Blobel,  G.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 293(19): 7160-7175.  2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.RA117.000678\"\n     onclick=\"log_download('wai-szyszka-campbell-kwong-lorna-silva-low-kwan-etal-thebrd3etdomainrecognizesashortpeptidemotifthroughamechanismthatisconservedacrosschromatinremodelersandtranscriptionalregulators-2018', 'http://doi.org/10.1074/jbc.RA117.000678', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wai-szyszka-campbell-kwong-lorna-silva-low-kwan-etal-thebrd3etdomainrecognizesashortpeptidemotifthroughamechanismthatisconservedacrosschromatinremodelersandtranscriptionalregulators-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wai-szyszka-campbell-kwong-lorna-silva-low-kwan-etal-thebrd3etdomainrecognizesashortpeptidemotifthroughamechanismthatisconservedacrosschromatinremodelersandtranscriptionalregulators-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The BRD3 ET domain recognizes a short peptide motif through a mechanism that is conserved across chromatin remodelers and transcriptional regulators},\n type = {article},\n year = {2018},\n pages = {7160-7175},\n volume = {293},\n id = {312373ac-f639-35cd-8df1-cd3bdb1cbb6f},\n created = {2023-01-10T01:44:13.874Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:13.874Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Members of the bromodomain and extra-terminal domain (BET) family of proteins (bromodomain-containing (BRD) 2, 3, 4, and T) are widely expressed and highly conserved regulators of gene expression in eukaryotes. These proteins have been intimately linked to human disease, and more than a dozen clinical trials are currently underway to test BET-protein inhibitors as modulators of cancer. However, although it is clear that these proteins use their bromodomains to bind both histones and transcription factors bearing acetylated lysine residues, the molecular mechanisms by which BET family proteins regulate gene expression are not well defined. In particular, the functions of the other domains such as the ET domain have been less extensively studied. Here, we examine the properties of the ET domain of BRD3 as a protein/protein interaction module. Using a combination of pulldown and biophysical assays, we demonstrate that BRD3 binds to a range of chromatin-remodeling complexes, including the NuRD, BAF, and INO80 complexes, via a short linear “KIKL” motif in one of the complex subunits. NMR-based structural analysis revealed that, surprisingly, this mode of interaction is shared by the AF9 and ENL transcriptional coregulators that contain an acetyl-lysine-binding YEATS domain and regulate transcriptional elongation. This observation establishes a functional commonality between these two families of cancer-related transcriptional regulators. In summary, our data provide insight into the mechanisms by which BET family proteins might link chromatin acetylation to transcriptional outcomes and uncover an unexpected functional similarity between BET and YEATS family proteins.},\n bibtype = {article},\n author = {Wai, D.C.C. and Szyszka, T.N. and Campbell, A.E. and Kwong, C. and Lorna, E.W.-W. and Silva, A.P.G. and Low, J.K.K. and Kwan, A.H. and Gamsjaeger, R. and Chalmers, J.D. and Blobel, G.A. and Mackay, J.P.},\n doi = {10.1074/jbc.RA117.000678},\n journal = {Journal of Biological Chemistry},\n number = {19}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Members of the bromodomain and extra-terminal domain (BET) family of proteins (bromodomain-containing (BRD) 2, 3, 4, and T) are widely expressed and highly conserved regulators of gene expression in eukaryotes. These proteins have been intimately linked to human disease, and more than a dozen clinical trials are currently underway to test BET-protein inhibitors as modulators of cancer. However, although it is clear that these proteins use their bromodomains to bind both histones and transcription factors bearing acetylated lysine residues, the molecular mechanisms by which BET family proteins regulate gene expression are not well defined. In particular, the functions of the other domains such as the ET domain have been less extensively studied. Here, we examine the properties of the ET domain of BRD3 as a protein/protein interaction module. Using a combination of pulldown and biophysical assays, we demonstrate that BRD3 binds to a range of chromatin-remodeling complexes, including the NuRD, BAF, and INO80 complexes, via a short linear “KIKL” motif in one of the complex subunits. NMR-based structural analysis revealed that, surprisingly, this mode of interaction is shared by the AF9 and ENL transcriptional coregulators that contain an acetyl-lysine-binding YEATS domain and regulate transcriptional elongation. This observation establishes a functional commonality between these two families of cancer-related transcriptional regulators. In summary, our data provide insight into the mechanisms by which BET family proteins might link chromatin acetylation to transcriptional outcomes and uncover an unexpected functional similarity between BET and YEATS family proteins.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"lu-klingbeil-tarumoto-somerville-huang-wei-wai-low-etal-atranscriptionfactoraddictioninleukemiaimposedbythemllpromotersequence-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A Transcription Factor Addiction in Leukemia Imposed by the MLL Promoter Sequence.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Lu,  B.; Klingbeil,  O.; Tarumoto,  Y.; Somerville,  T.; Huang,  Y.; Wei,  Y.; Wai,  D.; Low,  J.; Milazzo,  J.; Wu,  X.; Shi,  J.;  and Vakoc,  C.\n</span>\n\n  \n\n</span>\n\n  <i>Cancer Cell</i>, 34(6): 970-981.e8.  2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.ccell.2018.10.015\"\n     onclick=\"log_download('lu-klingbeil-tarumoto-somerville-huang-wei-wai-low-etal-atranscriptionfactoraddictioninleukemiaimposedbythemllpromotersequence-2018', 'http://doi.org/10.1016/j.ccell.2018.10.015', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/lu-klingbeil-tarumoto-somerville-huang-wei-wai-low-etal-atranscriptionfactoraddictioninleukemiaimposedbythemllpromotersequence-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('lu-klingbeil-tarumoto-somerville-huang-wei-wai-low-etal-atranscriptionfactoraddictioninleukemiaimposedbythemllpromotersequence-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A Transcription Factor Addiction in Leukemia Imposed by the MLL Promoter Sequence},\n type = {article},\n year = {2018},\n pages = {970-981.e8},\n volume = {34},\n id = {cdabe452-f0bc-3b9a-a76b-3912d34763ca},\n created = {2023-01-10T01:44:14.809Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:14.809Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The Mixed Lineage Leukemia gene (MLL) is altered in leukemia by chromosomal translocations to produce oncoproteins composed of the MLL N-terminus fused to the C-terminus of a partner protein. Here, we used domain-focused CRISPR screening to identify ZFP64 as an essential transcription factor in MLL-rearranged leukemia. We show that the critical function of ZFP64 in leukemia is to maintain MLL expression via binding to the MLL promoter, which is the most enriched location of ZFP64 occupancy in the human genome. The specificity of ZFP64 for MLL is accounted for by an exceptional density of ZFP64 motifs embedded within the MLL promoter. These findings demonstrate how a sequence anomaly of an oncogene promoter can impose a transcriptional addiction in cancer.SCOPUS_ABS_SEPARATORLu et al. show that MLL-rearranged leukemia is addicted to the transcription factor ZFP64 due to direct regulation of MLL by ZFP64. The MLL promoter has an unusually high number of ZFP64 binding motifs and is the most enriched location of ZFP64 occupancy in the human genome.},\n bibtype = {article},\n author = {Lu, B. and Klingbeil, O. and Tarumoto, Y. and Somerville, T.D.D. and Huang, Y.-H. and Wei, Y. and Wai, D.C. and Low, J.K.K. and Milazzo, J.P. and Wu, X.S. and Shi, J. and Vakoc, C.R.},\n doi = {10.1016/j.ccell.2018.10.015},\n journal = {Cancer Cell},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The Mixed Lineage Leukemia gene (MLL) is altered in leukemia by chromosomal translocations to produce oncoproteins composed of the MLL N-terminus fused to the C-terminus of a partner protein. Here, we used domain-focused CRISPR screening to identify ZFP64 as an essential transcription factor in MLL-rearranged leukemia. We show that the critical function of ZFP64 in leukemia is to maintain MLL expression via binding to the MLL promoter, which is the most enriched location of ZFP64 occupancy in the human genome. The specificity of ZFP64 for MLL is accounted for by an exceptional density of ZFP64 motifs embedded within the MLL promoter. These findings demonstrate how a sequence anomaly of an oncogene promoter can impose a transcriptional addiction in cancer.SCOPUS_ABS_SEPARATORLu et al. show that MLL-rearranged leukemia is addicted to the transcription factor ZFP64 due to direct regulation of MLL by ZFP64. The MLL promoter has an unusually high number of ZFP64 binding motifs and is the most enriched location of ZFP64 occupancy in the human genome.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Zhang,  X.; Farah,  N.; Rolston,  L.; Ericsson,  D.; Catanzariti,  A.; Bernoux,  M.; Ve,  T.; Bendak,  K.; Chen,  C.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Jones,  D.;  and Kobe,  B.\n</span>\n\n  \n\n</span>\n\n  <i>Molecular Plant Pathology</i>, 19(5): 1196-1209.  2018.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1111/mpp.12597\"\n     onclick=\"log_download('zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018', 'http://doi.org/10.1111/mpp.12597', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('zhang-farah-rolston-ericsson-catanzariti-bernoux-ve-bendak-etal-crystalstructureofthemelampsoralinieffectoravrprevealsinsightsintoapossiblenuclearfunctionandrecognitionbytheflaxdiseaseresistanceproteinp-2018')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Crystal structure of the Melampsora lini effector AvrP reveals insights into a possible nuclear function and recognition by the flax disease resistance protein P},\n type = {article},\n year = {2018},\n pages = {1196-1209},\n volume = {19},\n id = {baa2b413-e43a-312a-9a1b-c88fce658272},\n created = {2023-01-10T01:44:15.759Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:15.759Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.},\n bibtype = {article},\n author = {Zhang, X. and Farah, N. and Rolston, L. and Ericsson, D.J. and Catanzariti, A.-M. and Bernoux, M. and Ve, T. and Bendak, K. and Chen, C. and Mackay, J.P. and Jones, D.A. and Kobe, B.},\n doi = {10.1111/mpp.12597},\n journal = {Molecular Plant Pathology},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The effector protein AvrP is secreted by the flax rust fungal pathogen (Melampsora lini) and recognized specifically by the flax (Linum usitatissimum) P disease resistance protein, leading to effector-triggered immunity. To investigate the biological function of this effector and the mechanisms of specific recognition by the P resistance protein, we determined the crystal structure of AvrP. The structure reveals an elongated zinc-finger-like structure with a novel interleaved zinc-binding topology. The residues responsible for zinc binding are conserved in AvrP effector variants and mutations of these motifs result in a loss of P-mediated recognition. The first zinc-coordinating region of the structure displays a positively charged surface and shows some limited similarities to nucleic acid-binding and chromatin-associated proteins. We show that the majority of the AvrP protein accumulates in the plant nucleus when transiently expressed in Nicotiana benthamiana cells, suggesting a nuclear pathogenic function. Polymorphic residues in AvrP and its allelic variants map to the protein surface and could be associated with differences in recognition specificity. Several point mutations of residues on the non-conserved surface patch result in a loss of recognition by P, suggesting that these residues are required for recognition.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2017\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2017')\"\n      onkeypress=\"toggleGroup('group_2017')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2017</span>\n      <span class=\"bibbase_group_count\">\n        \n        (8)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1111/febs.14301\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017', 'http://doi.wiley.com/10.1111/febs.14301', event);\">\n    Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Torrado,  M.; Low,  J., K., K.; Silva,  A., P., G.; Schmidberger,  J., W.; Sana,  M.; Sharifi Tabar,  M.; Isilak,  M., E.; Winning,  C., S.; Kwong,  C.; Bedward,  M., J.; Sperlazza,  M., J.; Williams,  D., C.; Shepherd,  N., E.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>The FEBS Journal</i>, 284(24): 4216-4232. 12 2017.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1111/febs.14301\"\n     onclick=\"log_download('torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017', 'http://doi.wiley.com/10.1111/febs.14301', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1111/febs.14301\"\n     onclick=\"log_download('torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017', 'http://doi.org/10.1111/febs.14301', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex},\n type = {article},\n year = {2017},\n keywords = {chromatin remodelling,co-immunoprecipitations,nucleosome remodelling and deacetylase (NuRD) comp,protein structure,protein–protein interactions},\n pages = {4216-4232},\n volume = {284},\n websites = {http://doi.wiley.com/10.1111/febs.14301},\n month = {12},\n id = {fa40e576-8ce6-3e7e-83ec-453c57f0abe1},\n created = {2020-12-17T05:29:52.498Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:52.498Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Torrado2017},\n source_type = {ARTICLE},\n notes = {cited By 4},\n private_publication = {false},\n abstract = {The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.},\n bibtype = {article},\n author = {Torrado, Mario and Low, Jason K. K. and Silva, Ana P. G. and Schmidberger, Jason W. and Sana, Maryam and Sharifi Tabar, Mehdi and Isilak, Musa E. and Winning, Courtney S. and Kwong, Cherry and Bedward, Max J. and Sperlazza, Mary J. and Williams, David C. and Shepherd, Nicholas E. and Mackay, Joel P.},\n doi = {10.1111/febs.14301},\n journal = {The FEBS Journal},\n number = {24}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017', 'https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124', event);\">\n    Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Corcilius,  L.; Hastwell,  A., H.; Zhang,  M.; Williams,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Gresshoff,  P., M.; Ferguson,  B., J.;  and Payne,  R., J.\n</span>\n\n  \n\n</span>\n\n  <i>Cell Chemical Biology</i>, 24(11): 1347-1355.e7. 11 2017.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124\"\n     onclick=\"log_download('corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017', 'https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.chembiol.2017.08.014\"\n     onclick=\"log_download('corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017', 'http://doi.org/10.1016/j.chembiol.2017.08.014', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean},\n type = {article},\n year = {2017},\n keywords = {CLE,CLE40,IAD,arabinosylation,glycopeptide,glycosylation,hormone,legume,root apical meristem,soybean},\n pages = {1347-1355.e7},\n volume = {24},\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S2451945617303124},\n month = {11},\n id = {9da321e9-ae64-34bc-8487-6b2f91bcf82c},\n created = {2020-12-17T05:29:52.604Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:52.604Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Corcilius2017},\n source_type = {ARTICLE},\n notes = {cited By 8},\n private_publication = {false},\n abstract = {Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.},\n bibtype = {article},\n author = {Corcilius, Leo and Hastwell, April H. and Zhang, Mengbai and Williams, James and Mackay, Joel P. and Gresshoff, Peter M. and Ferguson, Brett J. and Payne, Richard J.},\n doi = {10.1016/j.chembiol.2017.08.014},\n journal = {Cell Chemical Biology},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Torrado,  M.; Low,  J.; Silva,  A.; Schmidberger,  J.; Sana,  M.; Sharifi Tabar,  M.; Isilak,  M.; Winning,  C.; Kwong,  C.; Bedward,  M.; Shepherd,  N.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>FEBS Journal</i>, 284(24): 4216-4232.  2017.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1111/febs.14301\"\n     onclick=\"log_download('torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017', 'http://doi.org/10.1111/febs.14301', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('torrado-low-silva-schmidberger-sana-sharifitabar-isilak-winning-etal-refinementofthesubunitinteractionnetworkwithinthenucleosomeremodellinganddeacetylasenurdcomplex-2017')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Refinement of the subunit interaction network within the nucleosome remodelling and deacetylase (NuRD) complex},\n type = {article},\n year = {2017},\n pages = {4216-4232},\n volume = {284},\n id = {125673fd-8a9b-3e40-83fe-795ba612fa34},\n created = {2023-01-10T01:44:16.686Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:16.686Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.},\n bibtype = {article},\n author = {Torrado, M. and Low, J.K.K. and Silva, A.P.G. and Schmidberger, J.W. and Sana, M. and Sharifi Tabar, M. and Isilak, M.E. and Winning, C.S. and Kwong, C. and Bedward, M.J. and Shepherd, N.E. and Mackay, J.P.},\n doi = {10.1111/febs.14301},\n journal = {FEBS Journal},\n number = {24}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes. Enzymes: Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Corcilius,  L.; Hastwell,  A.; Zhang,  M.; Williams,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Gresshoff,  P.; Ferguson,  B.;  and Payne,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Cell Chemical Biology</i>, 24(11): 1347-1355.e7.  2017.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.chembiol.2017.08.014\"\n     onclick=\"log_download('corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017', 'http://doi.org/10.1016/j.chembiol.2017.08.014', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('corcilius-hastwell-zhang-williams-mackay-gresshoff-ferguson-payne-arabinosylationmodulatesthegrowthregulatingactivityofthepeptidehormonecle40afromsoybean-2017')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Arabinosylation Modulates the Growth-Regulating Activity of the Peptide Hormone CLE40a from Soybean},\n type = {article},\n year = {2017},\n pages = {1347-1355.e7},\n volume = {24},\n id = {f5f29eda-8c79-33fb-a24e-f144305413fb},\n created = {2023-01-10T01:44:17.671Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:17.671Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.},\n bibtype = {article},\n author = {Corcilius, L. and Hastwell, A.H. and Zhang, M. and Williams, J. and Mackay, J.P. and Gresshoff, P.M. and Ferguson, B.J. and Payne, R.J.},\n doi = {10.1016/j.chembiol.2017.08.014},\n journal = {Cell Chemical Biology},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Small post-translationally modified peptide hormones mediate crucial developmental and regulatory processes in plants. CLAVATA/ENDOSPERM-SURROUNDING REGION (CLE) genes are found throughout the plant kingdom and encode for 12–13 amino acid peptides that must often undergo post-translational proline hydroxylation and glycosylation with O-β1,2-triarabinose moieties before they become functional. Apart from a few recent examples, a detailed understanding of the structure and function of most CLE hormones is yet to be uncovered. This is mainly owing to difficulties in isolating mature homogeneously modified CLE peptides from natural plant sources. In this study, we describe the efficient synthesis of a synthetic Araf 3 Hyp glycosylamino acid building block that was used to access a hitherto uninvestigated CLE hormone from soybean called GmCLE40a. Through the development and implementation of a novel in vivo root growth assay, we show that the synthetic triarabinosylated glycopeptide suppresses primary root growth in this important crop species. Herein Corcilius et al. describe an improved chemical synthesis of the β-O-tri-β1,2-arabinofuranosylated hydroxyproline post-translational modification present in plant glycopeptides and demonstrate its utility through the synthesis and biological evaluation of a novel root growth regulatory glycopeptide hormone, GmCLE40a, from soybean. The glycopeptide hormone displayed enhanced root growth suppressive activity in vivo compared with its unglycosylated isoform.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"ogeen-ren-nicolet-perez-halmai-le-mackay-farnham-etal-dcas9basedepigenomeeditingsuggestsacquisitionofhistonemethylationisnotsufficientfortargetgenerepression-2017\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  DCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  O'Geen,  H.; Ren,  C.; Nicolet,  C.; Perez,  A.; Halmai,  J.; Le,  V.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Farnham,  P.;  and Segal,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Nucleic Acids Research</i>, 45(17): 9901-9916.  2017.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1093/nar/gkx578\"\n     onclick=\"log_download('ogeen-ren-nicolet-perez-halmai-le-mackay-farnham-etal-dcas9basedepigenomeeditingsuggestsacquisitionofhistonemethylationisnotsufficientfortargetgenerepression-2017', 'http://doi.org/10.1093/nar/gkx578', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/ogeen-ren-nicolet-perez-halmai-le-mackay-farnham-etal-dcas9basedepigenomeeditingsuggestsacquisitionofhistonemethylationisnotsufficientfortargetgenerepression-2017\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('ogeen-ren-nicolet-perez-halmai-le-mackay-farnham-etal-dcas9basedepigenomeeditingsuggestsacquisitionofhistonemethylationisnotsufficientfortargetgenerepression-2017')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {DCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression},\n type = {article},\n year = {2017},\n pages = {9901-9916},\n volume = {45},\n id = {5eedcada-99ed-3068-9c49-72ae47c1066a},\n created = {2023-01-10T01:44:18.596Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:18.596Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Distinct epigenomic profiles of histone marks have been associated with gene expression, but questions regarding the causal relationship remain. Here we investigated the activity of a broad collection of genomically targeted epigenetic regulators that could write epigenetic marks associated with a repressed chromatin state (G9A, SUV39H1, Krüppel-associated box (KRAB), DNMT3A as well as the first targetable versions of Ezh2 and Friend of GATA-1 (FOG1)). dCas9 fusions produced target gene repression over a range of 0- to 10-fold that varied by locus and cell type. dCpf1 fusions were unable to repress gene expression. The most persistent gene repression required the action of several effector domains; however, KRAB-dCas9 did not contribute to persistence in contrast to previous reports. A &#x27;direct tethering&#x27; strategy attaching the Ezh2 methyltransferase enzyme to dCas9, as well as a &#x27;recruitment&#x27; strategy attaching the N-terminal 45 residues of FOG1 to dCas9 to recruit the endogenous nucleosome remodeling and deacetylase complex, were both successful in targeted deposition of H3K27me3. Surprisingly, however, repression was not correlated with deposition of either H3K9me3 or H3K27me3. Our results suggest that so-called repressive histone modifications are not sufficient for gene repression. The easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation.},\n bibtype = {article},\n author = {O&#x27;Geen, H. and Ren, C. and Nicolet, C.M. and Perez, A.A. and Halmai, J. and Le, V.M. and MacKay, J.P. and Farnham, P.J. and Segal, D.J.},\n doi = {10.1093/nar/gkx578},\n journal = {Nucleic Acids Research},\n number = {17}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Distinct epigenomic profiles of histone marks have been associated with gene expression, but questions regarding the causal relationship remain. Here we investigated the activity of a broad collection of genomically targeted epigenetic regulators that could write epigenetic marks associated with a repressed chromatin state (G9A, SUV39H1, Krüppel-associated box (KRAB), DNMT3A as well as the first targetable versions of Ezh2 and Friend of GATA-1 (FOG1)). dCas9 fusions produced target gene repression over a range of 0- to 10-fold that varied by locus and cell type. dCpf1 fusions were unable to repress gene expression. The most persistent gene repression required the action of several effector domains; however, KRAB-dCas9 did not contribute to persistence in contrast to previous reports. A 'direct tethering' strategy attaching the Ezh2 methyltransferase enzyme to dCas9, as well as a 'recruitment' strategy attaching the N-terminal 45 residues of FOG1 to dCas9 to recruit the endogenous nucleosome remodeling and deacetylase complex, were both successful in targeted deposition of H3K27me3. Surprisingly, however, repression was not correlated with deposition of either H3K9me3 or H3K27me3. Our results suggest that so-called repressive histone modifications are not sufficient for gene repression. The easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gillinder-ilsley-nbor-sachidanandam-lajoie-magor-tallack-bailey-etal-promiscuousdnabindingofamutantzincfingerproteincorruptsthetranscriptomeanddiminishescellviability-2017\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Promiscuous DNA-binding of a mutant zinc finger protein corrupts the transcriptome and diminishes cell viability.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gillinder,  K.; Ilsley,  M.; Nébor,  D.; Sachidanandam,  R.; Lajoie,  M.; Magor,  G.; Tallack,  M.; Bailey,  T.; Landsberg,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Bieker,  J.;  and Perkins,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Nucleic Acids Research</i>, 45(3): 1130-1143.  2017.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1093/nar/gkw1014\"\n     onclick=\"log_download('gillinder-ilsley-nbor-sachidanandam-lajoie-magor-tallack-bailey-etal-promiscuousdnabindingofamutantzincfingerproteincorruptsthetranscriptomeanddiminishescellviability-2017', 'http://doi.org/10.1093/nar/gkw1014', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gillinder-ilsley-nbor-sachidanandam-lajoie-magor-tallack-bailey-etal-promiscuousdnabindingofamutantzincfingerproteincorruptsthetranscriptomeanddiminishescellviability-2017\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>2 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gillinder-ilsley-nbor-sachidanandam-lajoie-magor-tallack-bailey-etal-promiscuousdnabindingofamutantzincfingerproteincorruptsthetranscriptomeanddiminishescellviability-2017')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Promiscuous DNA-binding of a mutant zinc finger protein corrupts the transcriptome and diminishes cell viability},\n type = {article},\n year = {2017},\n pages = {1130-1143},\n volume = {45},\n id = {fc32d854-4e6d-3528-9963-88946b3e8087},\n created = {2023-01-10T01:44:19.547Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:19.547Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The rules of engagement between zinc finger transcription factors and DNA have been partly defined by in vitro DNA-binding and structural studies, but less is known about how these rules apply in vivo. Here, we demonstrate how a missense mutation in the second zinc finger of Krüppel-like factor-1 (KLF1) leads to degenerate DNA-binding specificity in vivo, resulting in ectopic transcription and anemia in the Nanmouse model. We employed ChIP-seq and 4sU-RNA-seq to identify aberrant DNA-binding events genome wide and ectopic transcriptional consequences of this binding. We confirmed novel sequence specificity of the mutant recombinant zinc finger domain by performing biophysical measurements ofinvitroDNA-binding affinity. Together, these results shed new light on the mechanisms by which missense mutations in DNA-binding domains of transcription factors can lead to autosomal dominant diseases.},\n bibtype = {article},\n author = {Gillinder, K.R. and Ilsley, M.D. and Nébor, D. and Sachidanandam, R. and Lajoie, M. and Magor, G.W. and Tallack, M.R. and Bailey, T. and Landsberg, M.J. and Mackay, J.P. and Bieker, J.J. and Perkins, A.C.},\n doi = {10.1093/nar/gkw1014},\n journal = {Nucleic Acids Research},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The rules of engagement between zinc finger transcription factors and DNA have been partly defined by in vitro DNA-binding and structural studies, but less is known about how these rules apply in vivo. Here, we demonstrate how a missense mutation in the second zinc finger of Krüppel-like factor-1 (KLF1) leads to degenerate DNA-binding specificity in vivo, resulting in ectopic transcription and anemia in the Nanmouse model. We employed ChIP-seq and 4sU-RNA-seq to identify aberrant DNA-binding events genome wide and ectopic transcriptional consequences of this binding. We confirmed novel sequence specificity of the mutant recombinant zinc finger domain by performing biophysical measurements ofinvitroDNA-binding affinity. Together, these results shed new light on the mechanisms by which missense mutations in DNA-binding domains of transcription factors can lead to autosomal dominant diseases.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"li-lev-desmarini-kaufmanfrancis-saiardi-silva-mackay-thompson-etal-ipinf34infkinasearg1regulatescellwallhomeostasisandsurfacearchitecturetopromoteclearanceofcryptococcusneoformansinfectioninamousemodel-2017\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  IP<inf>3-4</inf> kinase Arg1 regulates cell wall homeostasis and surface architecture to promote clearance of Cryptococcus neoformans infection in a mouse model.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Li,  C.; Lev,  S.; Desmarini,  D.; Kaufman-Francis,  K.; Saiardi,  A.; Silva,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Thompson,  P.; Sorrell,  T.;  and Djordjevic,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Virulence</i>, 8(8): 1833-1848.  2017.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1080/21505594.2017.1385692\"\n     onclick=\"log_download('li-lev-desmarini-kaufmanfrancis-saiardi-silva-mackay-thompson-etal-ipinf34infkinasearg1regulatescellwallhomeostasisandsurfacearchitecturetopromoteclearanceofcryptococcusneoformansinfectioninamousemodel-2017', 'http://doi.org/10.1080/21505594.2017.1385692', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/li-lev-desmarini-kaufmanfrancis-saiardi-silva-mackay-thompson-etal-ipinf34infkinasearg1regulatescellwallhomeostasisandsurfacearchitecturetopromoteclearanceofcryptococcusneoformansinfectioninamousemodel-2017\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('li-lev-desmarini-kaufmanfrancis-saiardi-silva-mackay-thompson-etal-ipinf34infkinasearg1regulatescellwallhomeostasisandsurfacearchitecturetopromoteclearanceofcryptococcusneoformansinfectioninamousemodel-2017')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {IP&lt;inf&gt;3-4&lt;/inf&gt; kinase Arg1 regulates cell wall homeostasis and surface architecture to promote clearance of Cryptococcus neoformans infection in a mouse model},\n type = {article},\n year = {2017},\n pages = {1833-1848},\n volume = {8},\n id = {2ce6221b-c29d-38b5-826d-bcfd3df72bba},\n created = {2023-01-10T01:44:20.463Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:20.463Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {We previously identified a series of inositol polyphosphate kinases (IPKs), Arg1, Ipk1, Kcs1 and Asp1, in the opportunistic fungal pathogen Cryptococcus neoformans. Using gene deletion analysis, we characterized Arg1, Ipk1 and Kcs1 and showed that they act sequentially to convert IP3 to PP-IP5 (IP7), a key metabolite promoting stress tolerance, metabolic adaptation and fungal dissemination to the brain. We have now directly characterized the enzymatic activity of Arg1, demonstrating that it is a dual specificity (IP3/IP4) kinase producing IP5. We showed previously that IP5 is further phosphorylated by Ipk1 to produce IP6, which is a substrate for the synthesis of PP-IP5 by Kcs1. Phenotypic comparison of the arg1Δ and kcs1Δ deletion mutants (both PP-IP5-deficient) reveals that arg1Δ has the most deleterious phenotype: while PP-IP5 is essential for metabolic and stress adaptation in both mutant strains, PP-IP5 is dispensable for virulence-associated functions such as capsule production, cell wall organization, and normal N-linked mannosylation of the virulence factor, phospholipase B1, as these phenotypes were defective only in arg1Δ. The more deleterious arg1Δ phenotype correlated with a higher rate of arg1Δ phagocytosis by human peripheral blood monocytes and rapid arg1Δ clearance from lung in a mouse model. This observation is in contrast to kcs1Δ, which we previously reported establishes a chronic, confined lung infection. In summary, we show that Arg1 is the most crucial IPK for cryptococcal virulence, conveying PP-IP5–dependent and novel PP-IP5–independent functions.},\n bibtype = {article},\n author = {Li, C. and Lev, S. and Desmarini, D. and Kaufman-Francis, K. and Saiardi, A. and Silva, A.P.G. and Mackay, J.P. and Thompson, P.E. and Sorrell, T.C. and Djordjevic, J.T.},\n doi = {10.1080/21505594.2017.1385692},\n journal = {Virulence},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  We previously identified a series of inositol polyphosphate kinases (IPKs), Arg1, Ipk1, Kcs1 and Asp1, in the opportunistic fungal pathogen Cryptococcus neoformans. Using gene deletion analysis, we characterized Arg1, Ipk1 and Kcs1 and showed that they act sequentially to convert IP3 to PP-IP5 (IP7), a key metabolite promoting stress tolerance, metabolic adaptation and fungal dissemination to the brain. We have now directly characterized the enzymatic activity of Arg1, demonstrating that it is a dual specificity (IP3/IP4) kinase producing IP5. We showed previously that IP5 is further phosphorylated by Ipk1 to produce IP6, which is a substrate for the synthesis of PP-IP5 by Kcs1. Phenotypic comparison of the arg1Δ and kcs1Δ deletion mutants (both PP-IP5-deficient) reveals that arg1Δ has the most deleterious phenotype: while PP-IP5 is essential for metabolic and stress adaptation in both mutant strains, PP-IP5 is dispensable for virulence-associated functions such as capsule production, cell wall organization, and normal N-linked mannosylation of the virulence factor, phospholipase B1, as these phenotypes were defective only in arg1Δ. The more deleterious arg1Δ phenotype correlated with a higher rate of arg1Δ phagocytosis by human peripheral blood monocytes and rapid arg1Δ clearance from lung in a mouse model. This observation is in contrast to kcs1Δ, which we previously reported establishes a chronic, confined lung infection. In summary, we show that Arg1 is the most crucial IPK for cryptococcal virulence, conveying PP-IP5–dependent and novel PP-IP5–independent functions.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-landsberg-whitten-bond-whaddayaknowaguidetouncertaintyandsubjectivityinstructuralbiology-2017\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Whaddaya Know: A Guide to Uncertainty and Subjectivity in Structural Biology.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Landsberg,  M.; Whitten,  A.;  and Bond,  C.\n</span>\n\n  \n\n</span>\n\n  <i>Trends in Biochemical Sciences</i>, 42(2): 155-167.  2017.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.tibs.2016.11.002\"\n     onclick=\"log_download('mackay-landsberg-whitten-bond-whaddayaknowaguidetouncertaintyandsubjectivityinstructuralbiology-2017', 'http://doi.org/10.1016/j.tibs.2016.11.002', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-landsberg-whitten-bond-whaddayaknowaguidetouncertaintyandsubjectivityinstructuralbiology-2017\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-landsberg-whitten-bond-whaddayaknowaguidetouncertaintyandsubjectivityinstructuralbiology-2017')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Whaddaya Know: A Guide to Uncertainty and Subjectivity in Structural Biology},\n type = {article},\n year = {2017},\n pages = {155-167},\n volume = {42},\n id = {cea96057-95a9-35d5-851d-39b4311c3896},\n created = {2023-01-10T01:44:21.385Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:21.385Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The methods of structural biology, while powerful, are technically complex. Although the Protein Data Bank (PDB) provides a repository that allows anyone to download any structure, many users would not appreciate the caveats that should be considered when examining a structure. Here, we describe several key uncertainties associated with the application of X-ray crystallography, NMR spectroscopy, single-particle electron microscopy (SPEM), and small-angle scattering (SAS) to biological macromolecules. The take-home message is that structures are not absolute truths – they are models that fit the experimental data and therefore have uncertainty and subjectivity associated with them. These uncertainties must be appreciated – careful reading of the associated paper, and any validation report provided by the structure database, is highly recommended.},\n bibtype = {article},\n author = {Mackay, J.P. and Landsberg, M.J. and Whitten, A.E. and Bond, C.S.},\n doi = {10.1016/j.tibs.2016.11.002},\n journal = {Trends in Biochemical Sciences},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The methods of structural biology, while powerful, are technically complex. Although the Protein Data Bank (PDB) provides a repository that allows anyone to download any structure, many users would not appreciate the caveats that should be considered when examining a structure. Here, we describe several key uncertainties associated with the application of X-ray crystallography, NMR spectroscopy, single-particle electron microscopy (SPEM), and small-angle scattering (SAS) to biological macromolecules. The take-home message is that structures are not absolute truths – they are models that fit the experimental data and therefore have uncertainty and subjectivity associated with them. These uncertainties must be appreciated – careful reading of the associated paper, and any validation report provided by the structure database, is highly recommended.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2016\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2016')\"\n      onkeypress=\"toggleGroup('group_2016')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2016</span>\n      <span class=\"bibbase_group_count\">\n        \n        (13)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016', 'https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485', event);\">\n    The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mohanty,  B.; Helder,  S.; Silva,  A., P.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Ryan,  D., P.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 428(21): 4298-4314. 10 2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485\"\n     onclick=\"log_download('mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016', 'https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2016.08.028\"\n     onclick=\"log_download('mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016', 'http://doi.org/10.1016/j.jmb.2016.08.028', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain},\n type = {article},\n year = {2016},\n keywords = {C17orf64,DNA-binding domain,helical bundle,nucleosomes,transcription},\n pages = {4298-4314},\n volume = {428},\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S0022283616303485},\n month = {10},\n id = {851b1fbc-10b7-3289-b546-77e83ffb0c9d},\n created = {2020-12-17T05:29:53.207Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:53.207Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Mohanty2016},\n source_type = {ARTICLE},\n notes = {cited By 5},\n private_publication = {false},\n abstract = {The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.},\n bibtype = {article},\n author = {Mohanty, Biswaranjan and Helder, Stephanie and Silva, Ana P.G. and Mackay, Joel P. and Ryan, Daniel P.},\n doi = {10.1016/j.jmb.2016.08.028},\n journal = {Journal of Molecular Biology},\n number = {21}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1002/pro.2976\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016', 'http://doi.wiley.com/10.1002/pro.2976', event);\">\n    The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Blythe,  A., J.; Yazar-Klosinski,  B.; Webster,  M., W.; Chen,  E.; Vandevenne,  M.; Bendak,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Hartzog,  G., A.;  and Vrielink,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 25(9): 1710-1721. 9 2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1002/pro.2976\"\n     onclick=\"log_download('blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016', 'http://doi.wiley.com/10.1002/pro.2976', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.2976\"\n     onclick=\"log_download('blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016', 'http://doi.org/10.1002/pro.2976', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein},\n type = {article},\n year = {2016},\n keywords = {RNA binding,RNA polymerase,SELEX,Spt4/5,transcription elongation,transcription elongation factor},\n pages = {1710-1721},\n volume = {25},\n websites = {http://doi.wiley.com/10.1002/pro.2976},\n month = {9},\n id = {cb1f5362-a6b4-3c3e-a896-fd523c0b1404},\n created = {2020-12-17T05:29:56.143Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:56.143Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Blythe2016},\n source_type = {ARTICLE},\n notes = {cited By 2},\n private_publication = {false},\n abstract = {The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.},\n bibtype = {article},\n author = {Blythe, Amanda J. and Yazar-Klosinski, Berra and Webster, Michael W. and Chen, Eefei and Vandevenne, Marylène and Bendak, Katerina and Mackay, Joel P. and Hartzog, Grant A. and Vrielink, Alice},\n doi = {10.1002/pro.2976},\n journal = {Protein Science},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1002/pro.2943\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016', 'http://doi.wiley.com/10.1002/pro.2943', event);\">\n    The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Schmidberger,  J., W.; Sharifi Tabar,  M.; Torrado,  M.; Silva,  A., P., G.; Landsberg,  M., J.; Brillault,  L.; AlQarni,  S.; Zeng,  Y., C.; Parker,  B., L.; Low,  J., K., K.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 25(8): 1472-1482. 8 2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1002/pro.2943\"\n     onclick=\"log_download('schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016', 'http://doi.wiley.com/10.1002/pro.2943', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7 [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.2943\"\n     onclick=\"log_download('schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016', 'http://doi.org/10.1002/pro.2943', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7},\n type = {article},\n year = {2016},\n keywords = {MTA1,NuRD complex,RBBP4,chromatin,protein structure,transcription regulation},\n pages = {1472-1482},\n volume = {25},\n websites = {http://doi.wiley.com/10.1002/pro.2943},\n month = {8},\n id = {793f3c6b-8404-3b6e-9502-bf9b0357d1ca},\n created = {2020-12-17T05:29:59.209Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:59.209Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Schmidberger2016},\n source_type = {ARTICLE},\n notes = {cited By 4},\n private_publication = {false},\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.},\n bibtype = {article},\n author = {Schmidberger, Jason W. and Sharifi Tabar, Mehdi and Torrado, Mario and Silva, Ana P. G. and Landsberg, Michael J. and Brillault, Lou and AlQarni, Saad and Zeng, Yi Cheng and Parker, Benjamin L. and Low, Jason K. K. and Mackay, Joel P.},\n doi = {10.1002/pro.2943},\n journal = {Protein Science},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mohanty,  B.; Helder,  S.; Silva,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Ryan,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 428(21): 4298-4314.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2016.08.028\"\n     onclick=\"log_download('mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016', 'http://doi.org/10.1016/j.jmb.2016.08.028', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mohanty-helder-silva-mackay-ryan-thechromatinremodellingproteinchd1containsapreviouslyunrecognisedcterminalhelicaldomain-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The Chromatin Remodelling Protein CHD1 Contains a Previously Unrecognised C-Terminal Helical Domain},\n type = {article},\n year = {2016},\n pages = {4298-4314},\n volume = {428},\n id = {77987d3c-291d-39e9-97bc-2d692f6cd0c0},\n created = {2023-01-10T01:44:22.294Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:22.294Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.},\n bibtype = {article},\n author = {Mohanty, B. and Helder, S. and Silva, A.P.G. and Mackay, J.P. and Ryan, D.P.},\n doi = {10.1016/j.jmb.2016.08.028},\n journal = {Journal of Molecular Biology},\n number = {21}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The packaging of eukaryotic DNA into nucleosomes, and the organisation of these nucleosomes into chromatin, plays a critical role in regulating all DNA-associated processes. Chromodomain helicase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodelling protein that is conserved throughout eukaryotes and has an ability to assemble and organise nucleosomes both in vitro and in vivo. This activity is involved in the regulation of transcription and is implicated in mammalian development and stem cell biology. CHD1 is classically depicted as possessing a pair of tandem chromodomains that directly precede a core catalytic helicase-like domain that is then followed by a SANT-SLIDE DNA-binding domain. Here, we have identified an additional conserved domain C-terminal to the SANT-SLIDE domain and determined its structure by multidimensional heteronuclear NMR spectroscopy. We have termed this domain the CHD1 helical C-terminal (CHCT) domain as it is comprised of five α-helices arranged in a variant helical bundle topology. CHCT has a conserved, positively charged surface and is able to bind DNA and nucleosomes. In addition, we have identified another group of proteins, the as yet uncharacterised C17orf64 proteins, as also containing a conserved CHCT domain. Our data provide new structural insights into the CHD1 enzyme family.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"ittner-chua-bertz-volkerling-vanderhoven-gladbach-przybyla-bi-etal-sitespecificphosphorylationoftauinhibitsamyloidtoxicityinalzheimersmice-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Site-specific phosphorylation of tau inhibits amyloid-β toxicity in Alzheimer's mice.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Ittner,  A.; Chua,  S.; Bertz,  J.; Volkerling,  A.; Van Der Hoven,  J.; Gladbach,  A.; Przybyla,  M.; Bi,  M.; Van Hummel,  A.; Stevens,  C.; Ke,  Y.;  and Ittner,  L.\n</span>\n\n  \n\n</span>\n\n  <i>Science</i>, 354(6314): 904-908.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1126/science.aah6205\"\n     onclick=\"log_download('ittner-chua-bertz-volkerling-vanderhoven-gladbach-przybyla-bi-etal-sitespecificphosphorylationoftauinhibitsamyloidtoxicityinalzheimersmice-2016', 'http://doi.org/10.1126/science.aah6205', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/ittner-chua-bertz-volkerling-vanderhoven-gladbach-przybyla-bi-etal-sitespecificphosphorylationoftauinhibitsamyloidtoxicityinalzheimersmice-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('ittner-chua-bertz-volkerling-vanderhoven-gladbach-przybyla-bi-etal-sitespecificphosphorylationoftauinhibitsamyloidtoxicityinalzheimersmice-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Site-specific phosphorylation of tau inhibits amyloid-β toxicity in Alzheimer&#x27;s mice},\n type = {article},\n year = {2016},\n pages = {904-908},\n volume = {354},\n id = {569c0c3a-417c-3254-a9b2-e49317df57d6},\n created = {2023-01-10T01:44:23.226Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:23.226Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Amyloid-β (Aβ) toxicity in Alzheimer&#x27;s disease (AD) is considered to be mediated by phosphorylated tau protein. In contrast, we found that, at least in early disease, site-specific phosphorylation of tau inhibited Aβ toxicity. This specific tau phosphorylation was mediated by the neuronal p38 mitogen-activated protein kinase p38γ and interfered with postsynaptic excitotoxic signaling complexes engaged by Aβ. Accordingly, depletion of p38γ exacerbated neuronal circuit aberrations, cognitive deficits, and premature lethality in a mousemodel of AD, whereas increasing the activity of p38γ abolished these deficits. Furthermore, mimicking site-specific tau phosphorylation alleviated Aβ-induced neuronal death and offered protection from excitotoxicity. Our work provides insights into postsynaptic processes in AD pathogenesis and challenges a purely pathogenic role of tau phosphorylation in neuronal toxicity.},\n bibtype = {article},\n author = {Ittner, A. and Chua, S.W. and Bertz, J. and Volkerling, A. and Van Der Hoven, J. and Gladbach, A. and Przybyla, M. and Bi, M. and Van Hummel, A. and Stevens, C.H. and Ke, Y.D. and Ittner, L.M.},\n doi = {10.1126/science.aah6205},\n journal = {Science},\n number = {6314}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Amyloid-β (Aβ) toxicity in Alzheimer's disease (AD) is considered to be mediated by phosphorylated tau protein. In contrast, we found that, at least in early disease, site-specific phosphorylation of tau inhibited Aβ toxicity. This specific tau phosphorylation was mediated by the neuronal p38 mitogen-activated protein kinase p38γ and interfered with postsynaptic excitotoxic signaling complexes engaged by Aβ. Accordingly, depletion of p38γ exacerbated neuronal circuit aberrations, cognitive deficits, and premature lethality in a mousemodel of AD, whereas increasing the activity of p38γ abolished these deficits. Furthermore, mimicking site-specific tau phosphorylation alleviated Aβ-induced neuronal death and offered protection from excitotoxicity. Our work provides insights into postsynaptic processes in AD pathogenesis and challenges a purely pathogenic role of tau phosphorylation in neuronal toxicity.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wai-shihab-low-mackay-thezincfingersofyy1bindsinglestrandedrnawithlowsequencespecificity-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The zinc fingers of YY1 bind single-stranded RNA with low sequence specificity.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wai,  D.; Shihab,  M.; Low,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Nucleic Acids Research</i>, 44(19): 9153-9165.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1093/nar/gkw590\"\n     onclick=\"log_download('wai-shihab-low-mackay-thezincfingersofyy1bindsinglestrandedrnawithlowsequencespecificity-2016', 'http://doi.org/10.1093/nar/gkw590', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wai-shihab-low-mackay-thezincfingersofyy1bindsinglestrandedrnawithlowsequencespecificity-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wai-shihab-low-mackay-thezincfingersofyy1bindsinglestrandedrnawithlowsequencespecificity-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The zinc fingers of YY1 bind single-stranded RNA with low sequence specificity},\n type = {article},\n year = {2016},\n pages = {9153-9165},\n volume = {44},\n id = {448ec1cd-9b90-31ca-8df8-2d37eb36ba76},\n created = {2023-01-10T01:44:24.149Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:24.149Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Classical zinc fingers (ZFs) are traditionally considered to act as sequence-specific DNA-binding domains. More recently, classical ZFs have been recognised as potential RNA-binding modules, raising the intriguing possibility that classical-ZF transcription factors are involved in post-transcriptional gene regulation via direct RNA binding. To date, however, only one classical ZF-RNA complex, that involving TFIIIA, has been structurally characterised. Yin Yang-1 (YY1) is a multi-functional transcription factor involved in many regulatory processes, and binds DNA via four classical ZFs. Recent evidence suggests that YY1 also interacts with RNA, but the molecular nature of the interaction remains unknown. In the present work, we directly assess the ability of YY1 to bind RNA using in vitro assays. Systematic Evolution of Ligands by EXponential enrichment (SELEX) was used to identify preferred RNA sequences bound by the YY1 ZFs from a randomised library over multiple rounds of selection. However, a strong motif was not consistently recovered, suggesting that the RNA sequence selectivity of these domains is modest. YY1 ZF residues involved in binding to single-stranded RNA were identified by NMR spectroscopy and found to be largely distinct from the set of residues involved in DNA binding, suggesting that interactions between YY1 and ssRNA constitute a separate mode of nucleic acid binding. Our data are consistent with recent reports that YY1 can bind to RNA in a low-specificity, yet physiologically relevant manner.},\n bibtype = {article},\n author = {Wai, D.C.C. and Shihab, M. and Low, J.K.K. and Mackay, J.P.},\n doi = {10.1093/nar/gkw590},\n journal = {Nucleic Acids Research},\n number = {19}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Classical zinc fingers (ZFs) are traditionally considered to act as sequence-specific DNA-binding domains. More recently, classical ZFs have been recognised as potential RNA-binding modules, raising the intriguing possibility that classical-ZF transcription factors are involved in post-transcriptional gene regulation via direct RNA binding. To date, however, only one classical ZF-RNA complex, that involving TFIIIA, has been structurally characterised. Yin Yang-1 (YY1) is a multi-functional transcription factor involved in many regulatory processes, and binds DNA via four classical ZFs. Recent evidence suggests that YY1 also interacts with RNA, but the molecular nature of the interaction remains unknown. In the present work, we directly assess the ability of YY1 to bind RNA using in vitro assays. Systematic Evolution of Ligands by EXponential enrichment (SELEX) was used to identify preferred RNA sequences bound by the YY1 ZFs from a randomised library over multiple rounds of selection. However, a strong motif was not consistently recovered, suggesting that the RNA sequence selectivity of these domains is modest. YY1 ZF residues involved in binding to single-stranded RNA were identified by NMR spectroscopy and found to be largely distinct from the set of residues involved in DNA binding, suggesting that interactions between YY1 and ssRNA constitute a separate mode of nucleic acid binding. Our data are consistent with recent reports that YY1 can bind to RNA in a low-specificity, yet physiologically relevant manner.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"helder-blythe-bond-mackay-determinantsofaffinityandspecificityinrnabindingproteins-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Determinants of affinity and specificity in RNA-binding proteins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Helder,  S.; Blythe,  A.; Bond,  C.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Current Opinion in Structural Biology</i>, 38: 83-91.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.sbi.2016.05.005\"\n     onclick=\"log_download('helder-blythe-bond-mackay-determinantsofaffinityandspecificityinrnabindingproteins-2016', 'http://doi.org/10.1016/j.sbi.2016.05.005', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/helder-blythe-bond-mackay-determinantsofaffinityandspecificityinrnabindingproteins-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('helder-blythe-bond-mackay-determinantsofaffinityandspecificityinrnabindingproteins-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Determinants of affinity and specificity in RNA-binding proteins},\n type = {article},\n year = {2016},\n pages = {83-91},\n volume = {38},\n id = {61fe4e83-b401-35f1-9433-78ccbe3d486b},\n created = {2023-01-10T01:44:25.084Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:25.084Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Emerging data suggest that the mechanisms by which RNA-binding proteins (RBPs) interact with RNA and the rules governing specificity might be substantially more complex than those underlying their DNA-binding counterparts. Even our knowledge of what constitutes the RNA-bound proteome is contentious; recent studies suggest that 10-30% of RBPs contain no known RNA-binding domain. Adding to this situation is a growing disconnect between the avalanche of identified interactions between proteins and long noncoding RNAs and the absence of biophysical data on these interactions. RNA-protein interactions are also at the centre of what might emerge as one of the biggest shifts in thinking about cell and molecular biology this century, following from recent reports of ribonucleoprotein complexes that drive reversible membrane-free phase separation events within the cell. Unexpectedly, low-complexity motifs are important in the formation of these structures. Here we briefly survey recent advances in our understanding of the specificity of RBPs.},\n bibtype = {article},\n author = {Helder, S. and Blythe, A.J. and Bond, C.S. and Mackay, J.P.},\n doi = {10.1016/j.sbi.2016.05.005},\n journal = {Current Opinion in Structural Biology}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Emerging data suggest that the mechanisms by which RNA-binding proteins (RBPs) interact with RNA and the rules governing specificity might be substantially more complex than those underlying their DNA-binding counterparts. Even our knowledge of what constitutes the RNA-bound proteome is contentious; recent studies suggest that 10-30% of RBPs contain no known RNA-binding domain. Adding to this situation is a growing disconnect between the avalanche of identified interactions between proteins and long noncoding RNAs and the absence of biophysical data on these interactions. RNA-protein interactions are also at the centre of what might emerge as one of the biggest shifts in thinking about cell and molecular biology this century, following from recent reports of ribonucleoprotein complexes that drive reversible membrane-free phase separation events within the cell. Unexpectedly, low-complexity motifs are important in the formation of these structures. Here we briefly survey recent advances in our understanding of the specificity of RBPs.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Schmidberger,  J.; Sharifi Tabar,  M.; Torrado,  M.; Silva,  A.; Landsberg,  M.; Brillault,  L.; AlQarni,  S.; Zeng,  Y.; Parker,  B.; Low,  J.; Low,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>,1472-1482.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.2943\"\n     onclick=\"log_download('schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016', 'http://doi.org/10.1002/pro.2943', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('schmidberger-sharifitabar-torrado-silva-landsberg-brillault-alqarni-zeng-etal-themta1subunitofthenucleosomeremodelinganddeacetylasecomplexcanrecruittwocopiesofrbbp47-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The MTA1 subunit of the nucleosome remodeling and deacetylase complex can recruit two copies of RBBP4/7},\n type = {article},\n year = {2016},\n pages = {1472-1482},\n id = {a9a74730-2adb-3cee-87b3-9643769fab42},\n created = {2023-01-10T01:44:26.953Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:26.953Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.},\n bibtype = {article},\n author = {Schmidberger, J.W. and Sharifi Tabar, M. and Torrado, M. and Silva, A.P.G. and Landsberg, M.J. and Brillault, L. and AlQarni, S. and Zeng, Y.C. and Parker, B.L. and Low, J.K.K. and Low, J.K.K. and Mackay, J.P.},\n doi = {10.1002/pro.2943},\n journal = {Protein Science}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nucleosome remodeling and deacetylase (NuRD) complex remodels the genome in the context of both gene transcription and DNA damage repair. It is essential for normal development and is distributed across multiple tissues in organisms ranging from mammals to nematode worms. In common with other chromatin-remodeling complexes, however, its molecular mechanism of action is not well understood and only limited structural information is available to show how the complex is assembled. As a step towards understanding the structure of the NuRD complex, we have characterized the interaction between two subunits: the metastasis associated protein MTA1 and the histone-binding protein RBBP4. We show that MTA1 can bind to two molecules of RBBP4 and present negative stain electron microscopy and chemical crosslinking data that allow us to build a low-resolution model of an MTA1-(RBBP4)2 subcomplex. These data build on our understanding of NuRD complex structure and move us closer towards an understanding of the biochemical basis for the activity of this complex.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"silva-ryan-galanty-low-vandevenne-jackson-mackay-thenterminalregionofchromodomainhelicasednabindingprotein4chd4isessentialforactivityandcontainsahighmobilitygrouphmgboxlikedomainthatcanbindpolyadpribose-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The N-terminal region of chromodomain helicase DNA-binding protein 4 (CHD4) is essential for activity and contains a high mobility group (HMG) box-like-domain that can bind poly(ADP-ribose).\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Silva,  A.; Ryan,  D.; Galanty,  Y.; Low,  J.; Vandevenne,  M.; Jackson,  S.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 291(2): 924-938.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M115.683227\"\n     onclick=\"log_download('silva-ryan-galanty-low-vandevenne-jackson-mackay-thenterminalregionofchromodomainhelicasednabindingprotein4chd4isessentialforactivityandcontainsahighmobilitygrouphmgboxlikedomainthatcanbindpolyadpribose-2016', 'http://doi.org/10.1074/jbc.M115.683227', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/silva-ryan-galanty-low-vandevenne-jackson-mackay-thenterminalregionofchromodomainhelicasednabindingprotein4chd4isessentialforactivityandcontainsahighmobilitygrouphmgboxlikedomainthatcanbindpolyadpribose-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('silva-ryan-galanty-low-vandevenne-jackson-mackay-thenterminalregionofchromodomainhelicasednabindingprotein4chd4isessentialforactivityandcontainsahighmobilitygrouphmgboxlikedomainthatcanbindpolyadpribose-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The N-terminal region of chromodomain helicase DNA-binding protein 4 (CHD4) is essential for activity and contains a high mobility group (HMG) box-like-domain that can bind poly(ADP-ribose)},\n type = {article},\n year = {2016},\n pages = {924-938},\n volume = {291},\n id = {ecf7907f-b86b-3542-ab23-e2a9a7ff5cc9},\n created = {2023-01-10T01:44:27.879Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:27.879Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Chromodomain Helicase DNA-binding protein 4 (CHD4) is a chromatin-remodeling enzyme that has been reported to regulate DNA-damage responses through its N-terminal region in a poly(ADP-ribose) polymerase-dependent manner. We have identified and determined the structure of a stable domain (CHD4-N) in this N-terminal region. The-fold consists of a four-α-helix bundle with structural similarity to the high mobility group box, a domain that is well known as a DNA binding module. We show that the CHD4-N domain binds with higher affinity to poly(ADP-ribose) than to DNA. We also show that the N-terminal region of CHD4, although not CHD4-N alone, is essential for full nucleosome remodeling activity and is important for localizing CHD4 to sites of DNA damage. Overall, these data build on our understanding of how CHD4-NuRD acts to regulate gene expression and participates in the DNA-damage response.},\n bibtype = {article},\n author = {Silva, A.P.G. and Ryan, D.P. and Galanty, Y. and Low, J.K.K. and Vandevenne, M. and Jackson, S.P. and Mackay, J.P.},\n doi = {10.1074/jbc.M115.683227},\n journal = {Journal of Biological Chemistry},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chromodomain Helicase DNA-binding protein 4 (CHD4) is a chromatin-remodeling enzyme that has been reported to regulate DNA-damage responses through its N-terminal region in a poly(ADP-ribose) polymerase-dependent manner. We have identified and determined the structure of a stable domain (CHD4-N) in this N-terminal region. The-fold consists of a four-α-helix bundle with structural similarity to the high mobility group box, a domain that is well known as a DNA binding module. We show that the CHD4-N domain binds with higher affinity to poly(ADP-ribose) than to DNA. We also show that the N-terminal region of CHD4, although not CHD4-N alone, is essential for full nucleosome remodeling activity and is important for localizing CHD4 to sites of DNA damage. Overall, these data build on our understanding of how CHD4-NuRD acts to regulate gene expression and participates in the DNA-damage response.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"low-webb-silva-saathoff-ryan-torrado-brofelth-parker-etal-chd4isaperipheralcomponentofthenucleosomeremodelinganddeacetylasecomplex-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  CHD4 is a peripheral component of the nucleosome remodeling and deacetylase complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Low,  J.; Webb,  S.; Silva,  A.; Saathoff,  H.; Ryan,  D.; Torrado,  M.; Brofelth,  M.; Parker,  B.; Shepherd,  N.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 291(30): 15853-15866.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M115.707018\"\n     onclick=\"log_download('low-webb-silva-saathoff-ryan-torrado-brofelth-parker-etal-chd4isaperipheralcomponentofthenucleosomeremodelinganddeacetylasecomplex-2016', 'http://doi.org/10.1074/jbc.M115.707018', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/low-webb-silva-saathoff-ryan-torrado-brofelth-parker-etal-chd4isaperipheralcomponentofthenucleosomeremodelinganddeacetylasecomplex-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('low-webb-silva-saathoff-ryan-torrado-brofelth-parker-etal-chd4isaperipheralcomponentofthenucleosomeremodelinganddeacetylasecomplex-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {CHD4 is a peripheral component of the nucleosome remodeling and deacetylase complex},\n type = {article},\n year = {2016},\n pages = {15853-15866},\n volume = {291},\n id = {7b57ef93-0acb-3843-a769-694abe573e25},\n created = {2023-01-10T01:44:28.938Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:28.938Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Chromatin remodeling enzymes act to dynamically regulate gene accessibility. In many cases, these enzymes function as large multicomponent complexes that in general comprise a central ATP-dependent Snf2 family helicase that is decorated with a variable number of regulatory subunits. The nucleosome remodeling and deacetylase (NuRD) complex, which is essential for normal development in higher organisms, is one such macromolecular machine. The NuRD complex comprises ∼10 subunits, including the histone deacetylases 1 and 2 (HDAC1 and HDAC2), and is defined by the presence of a CHD family remodeling enzyme, most commonly CHD4 (chromodomain helicase DNA-binding protein 4). The existing paradigm holds that CHD4 acts as the central hub upon which the complex is built. We show here that this paradigm does not, in fact, hold and that CHD4 is a peripheral component of the NuRD complex. A complex lacking CHD4 that has HDAC activity can exist as a stable species. The addition of recombinant CHD4 to this nucleosome deacetylase complex reconstitutes a NuRD complex with nucleosome remodeling activity. These data contribute to our understanding of the architecture of the NuRD complex.},\n bibtype = {article},\n author = {Low, J.K.K. and Webb, S.R. and Silva, A.P.G. and Saathoff, H. and Ryan, D.P. and Torrado, M. and Brofelth, M. and Parker, B.L. and Shepherd, N.E. and Mackay, J.P.},\n doi = {10.1074/jbc.M115.707018},\n journal = {Journal of Biological Chemistry},\n number = {30}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chromatin remodeling enzymes act to dynamically regulate gene accessibility. In many cases, these enzymes function as large multicomponent complexes that in general comprise a central ATP-dependent Snf2 family helicase that is decorated with a variable number of regulatory subunits. The nucleosome remodeling and deacetylase (NuRD) complex, which is essential for normal development in higher organisms, is one such macromolecular machine. The NuRD complex comprises ∼10 subunits, including the histone deacetylases 1 and 2 (HDAC1 and HDAC2), and is defined by the presence of a CHD family remodeling enzyme, most commonly CHD4 (chromodomain helicase DNA-binding protein 4). The existing paradigm holds that CHD4 acts as the central hub upon which the complex is built. We show here that this paradigm does not, in fact, hold and that CHD4 is a peripheral component of the NuRD complex. A complex lacking CHD4 that has HDAC activity can exist as a stable species. The addition of recombinant CHD4 to this nucleosome deacetylase complex reconstitutes a NuRD complex with nucleosome remodeling activity. These data contribute to our understanding of the architecture of the NuRD complex.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"luo-xue-kwan-gamsjaeger-wielens-vonkleist-cubeddu-guo-etal-thebindingofsyndapinsh3domaintodynaminprolinerichdomaininvolvesshortandlongdistanceelements-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The binding of syndapin SH3 domain to Dynamin proline-rich domain involves short and long distance elements.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Luo,  L.; Xue,  J.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Gamsjaeger,  R.; Wielens,  J.; Von Kleist,  L.; Cubeddu,  L.; Guo,  Z.; Stow,  J.; Parker,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Robinson,  P.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 291(18): 9411-9424.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M115.703108\"\n     onclick=\"log_download('luo-xue-kwan-gamsjaeger-wielens-vonkleist-cubeddu-guo-etal-thebindingofsyndapinsh3domaintodynaminprolinerichdomaininvolvesshortandlongdistanceelements-2016', 'http://doi.org/10.1074/jbc.M115.703108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/luo-xue-kwan-gamsjaeger-wielens-vonkleist-cubeddu-guo-etal-thebindingofsyndapinsh3domaintodynaminprolinerichdomaininvolvesshortandlongdistanceelements-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('luo-xue-kwan-gamsjaeger-wielens-vonkleist-cubeddu-guo-etal-thebindingofsyndapinsh3domaintodynaminprolinerichdomaininvolvesshortandlongdistanceelements-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The binding of syndapin SH3 domain to Dynamin proline-rich domain involves short and long distance elements},\n type = {article},\n year = {2016},\n pages = {9411-9424},\n volume = {291},\n id = {34917dfc-3a3c-379e-a481-47c891a62429},\n created = {2023-01-10T01:44:29.922Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:29.922Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Dynamin is a GTPase that mediates vesicle fission during synaptic vesicle endocytosis. Its long C-terminal proline-rich domain contains 13 PXXP motifs, which orchestrate its interactions with multiple proteins. The SH3 domains of syndapin and endophilin bind the PXXP motifs called Site 2 and 3 (Pro-786-Pro-793) at the N-terminal end of the proline-rich domain, whereas the amphiphysin SH3 binds Site 9 (Pro-833-Pro-836) toward the C-terminal end. In some proteins, SH3/peptide interactions also involve short distance elements, which are 5-15 amino acid extensions flanking the central PXXP motif for high affinity binding. Here we found two previously unrecognized elements in the central and the C-terminal end of the dynamin proline-rich domain that account for a significant increase in syndapin binding affinity compared with a previously reported Site 2 and Site 3 PXXP peptide alone. The first new element (Gly-807-Gly-811) is short distance element on the C-terminal side of Site 2 PXXP, which might contact a groove identified under the RT loop of the SH3 domain. The second element (Arg-838-Pro-844) is located about 50 amino acids downstream of Site 2. These two elements provide additional specificity tothe syndapin SH3 domain outsideofthe well described polyproline-binding groove. Thus, the dynamin/syndapin interaction is mediated via a network of multiple contacts outside the core PXXP motif over a previously unrecognized extended region of the proline-rich domain. To our knowledge this is the first example among known SH3 interactions to involve spatially separated and extended long-range elements that combine to provide a higher affinity interaction. 7copy;2016 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Luo, L. and Xue, J. and Kwan, A. and Gamsjaeger, R. and Wielens, J. and Von Kleist, L. and Cubeddu, L. and Guo, Z. and Stow, J.L. and Parker, M.W. and Mackay, J.P. and Robinson, P.J.},\n doi = {10.1074/jbc.M115.703108},\n journal = {Journal of Biological Chemistry},\n number = {18}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Dynamin is a GTPase that mediates vesicle fission during synaptic vesicle endocytosis. Its long C-terminal proline-rich domain contains 13 PXXP motifs, which orchestrate its interactions with multiple proteins. The SH3 domains of syndapin and endophilin bind the PXXP motifs called Site 2 and 3 (Pro-786-Pro-793) at the N-terminal end of the proline-rich domain, whereas the amphiphysin SH3 binds Site 9 (Pro-833-Pro-836) toward the C-terminal end. In some proteins, SH3/peptide interactions also involve short distance elements, which are 5-15 amino acid extensions flanking the central PXXP motif for high affinity binding. Here we found two previously unrecognized elements in the central and the C-terminal end of the dynamin proline-rich domain that account for a significant increase in syndapin binding affinity compared with a previously reported Site 2 and Site 3 PXXP peptide alone. The first new element (Gly-807-Gly-811) is short distance element on the C-terminal side of Site 2 PXXP, which might contact a groove identified under the RT loop of the SH3 domain. The second element (Arg-838-Pro-844) is located about 50 amino acids downstream of Site 2. These two elements provide additional specificity tothe syndapin SH3 domain outsideofthe well described polyproline-binding groove. Thus, the dynamin/syndapin interaction is mediated via a network of multiple contacts outside the core PXXP motif over a previously unrecognized extended region of the proline-rich domain. To our knowledge this is the first example among known SH3 interactions to involve spatially separated and extended long-range elements that combine to provide a higher affinity interaction. 7copy;2016 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mohanty-silva-mackay-ryan-sup1suphsup13supcandsup15supnresonanceassignmentsofacterminaldomainofhumanchd1-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of a C-terminal domain of human CHD1.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mohanty,  B.; Silva,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Ryan,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 10(1): 31-34.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-015-9631-1\"\n     onclick=\"log_download('mohanty-silva-mackay-ryan-sup1suphsup13supcandsup15supnresonanceassignmentsofacterminaldomainofhumanchd1-2016', 'http://doi.org/10.1007/s12104-015-9631-1', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mohanty-silva-mackay-ryan-sup1suphsup13supcandsup15supnresonanceassignmentsofacterminaldomainofhumanchd1-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mohanty-silva-mackay-ryan-sup1suphsup13supcandsup15supnresonanceassignmentsofacterminaldomainofhumanchd1-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {&lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;13&lt;/sup&gt;C and &lt;sup&gt;15&lt;/sup&gt;N resonance assignments of a C-terminal domain of human CHD1},\n type = {article},\n year = {2016},\n pages = {31-34},\n volume = {10},\n id = {b856848d-bf8e-37fe-813e-40d403bbd998},\n created = {2023-01-10T01:44:30.848Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:30.848Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Chromatin remodelling proteins are an essential family of eukaryotic proteins. They harness the energy from ATP hydrolysis and apply it to alter chromatin structure in order to regulate all aspects of genome biology. Chromodomain helicase DNA-binding protein 1 (CHD1) is one such remodelling protein that has specialised nucleosome organising abilities and is conserved across eukaryotes. CHD1 possesses a pair of tandem chromodomains that directly precede the core catalytic Snf2 helicase-like domain, and a C-terminal SANT-SLIDE DNA-binding domain. We have identified an additional conserved domain in the C-terminal region of CHD1. Here, we report the backbone and side chain resonance assignments for this domain from human CHD1 at pH 6.5 and 25 °C (BMRB No. 25638).},\n bibtype = {article},\n author = {Mohanty, B. and Silva, A.P.G. and Mackay, J.P. and Ryan, D.P.},\n doi = {10.1007/s12104-015-9631-1},\n journal = {Biomolecular NMR Assignments},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chromatin remodelling proteins are an essential family of eukaryotic proteins. They harness the energy from ATP hydrolysis and apply it to alter chromatin structure in order to regulate all aspects of genome biology. Chromodomain helicase DNA-binding protein 1 (CHD1) is one such remodelling protein that has specialised nucleosome organising abilities and is conserved across eukaryotes. CHD1 possesses a pair of tandem chromodomains that directly precede the core catalytic Snf2 helicase-like domain, and a C-terminal SANT-SLIDE DNA-binding domain. We have identified an additional conserved domain in the C-terminal region of CHD1. Here, we report the backbone and side chain resonance assignments for this domain from human CHD1 at pH 6.5 and 25 °C (BMRB No. 25638).\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Blythe,  A.; Yazar-Klosinski,  B.; Webster,  M.; Chen,  E.; Vandevenne,  M.; Bendak,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Hartzog,  G.;  and Vrielink,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>,1710-1721.  2016.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.2976\"\n     onclick=\"log_download('blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016', 'http://doi.org/10.1002/pro.2976', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('blythe-yazarklosinski-webster-chen-vandevenne-bendak-mackay-hartzog-etal-theyeasttranscriptionelongationfactorspt45isasequencespecificrnabindingprotein-2016')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein},\n type = {article},\n year = {2016},\n pages = {1710-1721},\n id = {7411c703-cf65-3456-a534-495980102c40},\n created = {2023-01-10T01:44:31.837Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:31.837Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.},\n bibtype = {article},\n author = {Blythe, A.J. and Yazar-Klosinski, B. and Webster, M.W. and Chen, E. and Vandevenne, M. and Bendak, K. and Mackay, J.P. and Hartzog, G.A. and Vrielink, A.},\n doi = {10.1002/pro.2976},\n journal = {Protein Science}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2015\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2015')\"\n      onkeypress=\"toggleGroup('group_2015')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2015</span>\n      <span class=\"bibbase_group_count\">\n        \n        (4)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015', 'https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395', event);\">\n    A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Saathoff,  H.; Brofelth,  M.; Trinh,  A.; Parker,  B., L.; Ryan,  D., P.; Low,  J., K.; Webb,  S., R.; Silva,  A., P.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Shepherd,  N., E.\n</span>\n\n  \n\n</span>\n\n  <i>Bioorganic & Medicinal Chemistry</i>, 23(5): 960-965. 3 2015.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395\"\n     onclick=\"log_download('saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015', 'https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.bmc.2015.01.023\"\n     onclick=\"log_download('saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015', 'http://doi.org/10.1016/j.bmc.2015.01.023', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells},\n type = {article},\n year = {2015},\n keywords = {Affinity purification,Friend of GATA1 (FOG1),Multi-protein complex,Nucleosome remodeling and deacetylase complex pept},\n pages = {960-965},\n volume = {23},\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S0968089615000395},\n month = {3},\n id = {c7a33bb8-06cd-300f-9f85-c79fbd926928},\n created = {2020-12-17T05:29:56.497Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:56.497Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Saathoff2015},\n source_type = {ARTICLE},\n notes = {cited By 5},\n private_publication = {false},\n abstract = {We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).},\n bibtype = {article},\n author = {Saathoff, Hinnerk and Brofelth, Mattias and Trinh, Anne and Parker, Benjamin L. and Ryan, Daniel P. and Low, Jason K.K. and Webb, Sarah R. and Silva, Ana P.G. and Mackay, Joel P. and Shepherd, Nicholas E.},\n doi = {10.1016/j.bmc.2015.01.023},\n journal = {Bioorganic &amp; Medicinal Chemistry},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"stevenson-rabbolini-beutler-chen-gabrielli-mackay-brighton-ward-etal-paristrousseauthrombocytopeniaisphenocopiedbytheautosomalrecessiveinheritanceofadnabindingdomainmutationinfli1-2015\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Paris-Trousseau thrombocytopenia is phenocopied by the autosomal recessive inheritance of a DNA-binding domain mutation in FLI1.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Stevenson,  W.; Rabbolini,  D.; Beutler,  L.; Chen,  Q.; Gabrielli,  S.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Brighton,  T.; Ward,  C.;  and Morel-Kopp,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Blood</i>, 126(17): 2027-2030.  2015.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1182/blood-2015-06-650887\"\n     onclick=\"log_download('stevenson-rabbolini-beutler-chen-gabrielli-mackay-brighton-ward-etal-paristrousseauthrombocytopeniaisphenocopiedbytheautosomalrecessiveinheritanceofadnabindingdomainmutationinfli1-2015', 'http://doi.org/10.1182/blood-2015-06-650887', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/stevenson-rabbolini-beutler-chen-gabrielli-mackay-brighton-ward-etal-paristrousseauthrombocytopeniaisphenocopiedbytheautosomalrecessiveinheritanceofadnabindingdomainmutationinfli1-2015\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('stevenson-rabbolini-beutler-chen-gabrielli-mackay-brighton-ward-etal-paristrousseauthrombocytopeniaisphenocopiedbytheautosomalrecessiveinheritanceofadnabindingdomainmutationinfli1-2015')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Paris-Trousseau thrombocytopenia is phenocopied by the autosomal recessive inheritance of a DNA-binding domain mutation in FLI1},\n type = {article},\n year = {2015},\n pages = {2027-2030},\n volume = {126},\n id = {9411fc17-2fe7-33f7-b1a3-3b0944dfec36},\n created = {2023-01-10T01:44:32.853Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:32.853Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Hemizygous deletion of a variable regionon chromosome 11q containing FLI1 causes an inherited platelet-related bleeding disorder in Paris-Trousseau thrombocytopenia and Jacobsen syndrome. These multisystem disorders are also characterized by heart anomalies, changes in facial structure, and intellectual disability. We have identified a consanguineous family with autosomal recessive inheritance of a bleeding disorder that mimics Paris-Trousseau thrombocytopenia but has no other features of the 11q23 deletion syndrome. Affected individuals inthis family have moderate thrombocytopenia; absent collagen-induced platelet aggregation; and large, fused α-granules in1%to 5% of circulating platelets. This phenotype was caused by a FLI1 homozygous c.970C&gt;T-point mutation that predicts an arginine-to-tryptophan substitution in the conserved ETS DNA-binding domain of FLI1. This mutation caused a transcription defect at the promoter of known FLI1 target genes GP6, GP9, and ITGA2B, as measured by luciferase assay in HEK293 cells, and decreased the expression of these target proteins in affected members of the family as measured by Western blotting of platelet lysates. This kindred suggests abnormalitiesin FLI1 as causative of Paris-Trousseau thrombocytopenia and confirms the important role of FLI1 in normal platelet development.},\n bibtype = {article},\n author = {Stevenson, W.S. and Rabbolini, D.J. and Beutler, L. and Chen, Q. and Gabrielli, S. and Mackay, J.P. and Brighton, T.A. and Ward, C.M. and Morel-Kopp, M.-C.},\n doi = {10.1182/blood-2015-06-650887},\n journal = {Blood},\n number = {17}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Hemizygous deletion of a variable regionon chromosome 11q containing FLI1 causes an inherited platelet-related bleeding disorder in Paris-Trousseau thrombocytopenia and Jacobsen syndrome. These multisystem disorders are also characterized by heart anomalies, changes in facial structure, and intellectual disability. We have identified a consanguineous family with autosomal recessive inheritance of a bleeding disorder that mimics Paris-Trousseau thrombocytopenia but has no other features of the 11q23 deletion syndrome. Affected individuals inthis family have moderate thrombocytopenia; absent collagen-induced platelet aggregation; and large, fused α-granules in1%to 5% of circulating platelets. This phenotype was caused by a FLI1 homozygous c.970C>T-point mutation that predicts an arginine-to-tryptophan substitution in the conserved ETS DNA-binding domain of FLI1. This mutation caused a transcription defect at the promoter of known FLI1 target genes GP6, GP9, and ITGA2B, as measured by luciferase assay in HEK293 cells, and decreased the expression of these target proteins in affected members of the family as measured by Western blotting of platelet lysates. This kindred suggests abnormalitiesin FLI1 as causative of Paris-Trousseau thrombocytopenia and confirms the important role of FLI1 in normal platelet development.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"shepherd-gamsjaeger-vandevenne-cubeddu-mackay-sitedirectednitroxidespinlabelingofoligonucleotidesfornmrandeprstudies-2015\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Site directed nitroxide spin labeling of oligonucleotides for NMR and EPR studies.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Shepherd,  N.; Gamsjaeger,  R.; Vandevenne,  M.; Cubeddu,  L.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Tetrahedron</i>, 71(5): 813-819.  2015.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.tet.2014.12.056\"\n     onclick=\"log_download('shepherd-gamsjaeger-vandevenne-cubeddu-mackay-sitedirectednitroxidespinlabelingofoligonucleotidesfornmrandeprstudies-2015', 'http://doi.org/10.1016/j.tet.2014.12.056', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/shepherd-gamsjaeger-vandevenne-cubeddu-mackay-sitedirectednitroxidespinlabelingofoligonucleotidesfornmrandeprstudies-2015\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('shepherd-gamsjaeger-vandevenne-cubeddu-mackay-sitedirectednitroxidespinlabelingofoligonucleotidesfornmrandeprstudies-2015')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Site directed nitroxide spin labeling of oligonucleotides for NMR and EPR studies},\n type = {article},\n year = {2015},\n pages = {813-819},\n volume = {71},\n id = {2ba001b8-f1dd-3136-90e2-56100f2c35ff},\n created = {2023-01-10T01:44:33.864Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:33.864Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Nitroxide labels are useful probes of biomolecule structure that can be detected by NMR and EPR spectroscopy. Although many methods exist for labeling oligonucleotides with nitroxides, most require reagents that are expensive or laborious to prepare. A simpler approach is described herein using commercially available phosphorothioate oligonucleotides and 2-(3-iodoacetamidomethyl)-PROXYL. We describe semi-optimized conditions for labeling DNA and RNA oligonucleotides and methodology for purifying and identifying these reagents by MALDI-MS. The nitroxide label showed some propensity to hydrolyze at high temperature and over prolonged periods at room temperature. Nitroxide-labeled DNA oligonucleotides gave characteristic EPR spectra and caused the disappearance of bound protein signals in 15N HSQC spectra consistent with the paramagnetic relaxation enhancement (PRE) effect.},\n bibtype = {article},\n author = {Shepherd, N.E. and Gamsjaeger, R. and Vandevenne, M. and Cubeddu, L. and Mackay, J.P.},\n doi = {10.1016/j.tet.2014.12.056},\n journal = {Tetrahedron},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Nitroxide labels are useful probes of biomolecule structure that can be detected by NMR and EPR spectroscopy. Although many methods exist for labeling oligonucleotides with nitroxides, most require reagents that are expensive or laborious to prepare. A simpler approach is described herein using commercially available phosphorothioate oligonucleotides and 2-(3-iodoacetamidomethyl)-PROXYL. We describe semi-optimized conditions for labeling DNA and RNA oligonucleotides and methodology for purifying and identifying these reagents by MALDI-MS. The nitroxide label showed some propensity to hydrolyze at high temperature and over prolonged periods at room temperature. Nitroxide-labeled DNA oligonucleotides gave characteristic EPR spectra and caused the disappearance of bound protein signals in 15N HSQC spectra consistent with the paramagnetic relaxation enhancement (PRE) effect.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Saathoff,  H.; Brofelth,  M.; Trinh,  A.; Parker,  B.; Ryan,  D.; Low,  J.; Webb,  S.; Silva,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Shepherd,  N.\n</span>\n\n  \n\n</span>\n\n  <i>Bioorganic and Medicinal Chemistry</i>, 23(5): 960-965.  2015.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.bmc.2015.01.023\"\n     onclick=\"log_download('saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015', 'http://doi.org/10.1016/j.bmc.2015.01.023', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('saathoff-brofelth-trinh-parker-ryan-low-webb-silva-etal-apeptideaffinityreagentforisolatinganintactandcatalyticallyactivemultiproteincomplexfrommammaliancells-2015')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A peptide affinity reagent for isolating an intact and catalytically active multi-protein complex from mammalian cells},\n type = {article},\n year = {2015},\n pages = {960-965},\n volume = {23},\n id = {4e5561f0-05d0-3b6a-8a77-9e05d0b36591},\n created = {2023-01-10T01:44:34.787Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:34.787Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).},\n bibtype = {article},\n author = {Saathoff, H. and Brofelth, M. and Trinh, A. and Parker, B.L. and Ryan, D.P. and Low, J.K.K. and Webb, S.R. and Silva, A.P.G. and Mackay, J.P. and Shepherd, N.E.},\n doi = {10.1016/j.bmc.2015.01.023},\n journal = {Bioorganic and Medicinal Chemistry},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  We have developed an approach for directly isolating an intact multi-protein chromatin remodeling complex from mammalian cell extracts using synthetic peptide affinity reagent 4. FOG1(1-15), a short peptide sequence known to target subunits of the nucleosome remodeling and deacetylase (NuRD) complex, was joined via a 35-atom hydrophilic linker to the StreptagII peptide. Loading this peptide onto Streptactin beads enabled capture of the intact NuRD complex from MEL cell nuclear extract. Gentle biotin elution yielded the desired intact complex free of significant contaminants and in a form that was catalytically competent in a nucleosome remodeling assay. The efficiency of 4 in isolating the NuRD complex was comparable to other reported methods utilising recombinantly produced GST-FOG1(1-45).\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2014\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2014')\"\n      onkeypress=\"toggleGroup('group_2014')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2014</span>\n      <span class=\"bibbase_group_count\">\n        \n        (15)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"silva-kwan-mackay-1h13cand15nresonanceassignmentsofannterminaldomainofchd4-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://link.springer.com/10.1007/s12104-013-9469-3\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'silva-kwan-mackay-1h13cand15nresonanceassignmentsofannterminaldomainofchd4-2014', 'http://link.springer.com/10.1007/s12104-013-9469-3', event);\">\n    1H, 13C and 15N resonance assignments of an N-terminal domain of CHD4.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Silva,  A., P., G.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H.</a>;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 8(1): 137-139. 4 2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://link.springer.com/10.1007/s12104-013-9469-3\"\n     onclick=\"log_download('silva-kwan-mackay-1h13cand15nresonanceassignmentsofannterminaldomainofchd4-2014', 'http://link.springer.com/10.1007/s12104-013-9469-3', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"1H, 13C and 15N resonance assignments of an N-terminal domain of CHD4 [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-013-9469-3\"\n     onclick=\"log_download('silva-kwan-mackay-1h13cand15nresonanceassignmentsofannterminaldomainofchd4-2014', 'http://doi.org/10.1007/s12104-013-9469-3', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/silva-kwan-mackay-1h13cand15nresonanceassignmentsofannterminaldomainofchd4-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('silva-kwan-mackay-1h13cand15nresonanceassignmentsofannterminaldomainofchd4-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {1H, 13C and 15N resonance assignments of an N-terminal domain of CHD4},\n type = {article},\n year = {2014},\n keywords = {CHD4,Chromatin remodeling,N-terminal domain,NuRD complex,PAR-binding motif},\n pages = {137-139},\n volume = {8},\n websites = {http://link.springer.com/10.1007/s12104-013-9469-3},\n month = {4},\n day = {17},\n id = {6b604bfa-8cf1-39d7-9959-cdd1ad06e0c1},\n created = {2020-12-17T05:29:52.354Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:52.354Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Silva2014},\n source_type = {ARTICLE},\n notes = {cited By 1},\n private_publication = {false},\n abstract = {Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.},\n bibtype = {article},\n author = {Silva, Ana P. G. and Kwan, Ann H. and Mackay, Joel P.},\n doi = {10.1007/s12104-013-9469-3},\n journal = {Biomolecular NMR Assignments},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1002/anie.201402980\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014', 'http://doi.wiley.com/10.1002/anie.201402980', event);\">\n    Engineering Specificity Changes on a RanBP2 Zinc Finger that Binds Single-Stranded RNA.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Vandevenne,  M.; O'Connell,  M., R.; Helder,  S.; Shepherd,  N., E.; Matthews,  J., M.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H.</a>; Segal,  D., J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Angewandte Chemie International Edition</i>, 53(30): 7848-7852. 7 2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1002/anie.201402980\"\n     onclick=\"log_download('vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014', 'http://doi.wiley.com/10.1002/anie.201402980', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Engineering Specificity Changes on a RanBP2 Zinc Finger that Binds Single-Stranded RNA [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/anie.201402980\"\n     onclick=\"log_download('vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014', 'http://doi.org/10.1002/anie.201402980', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Engineering Specificity Changes on a RanBP2 Zinc Finger that Binds Single-Stranded RNA},\n type = {article},\n year = {2014},\n keywords = {RNA binding,combinatorial chemistry,phage display,protein design,zinc fingers},\n pages = {7848-7852},\n volume = {53},\n websites = {http://doi.wiley.com/10.1002/anie.201402980},\n month = {7},\n day = {21},\n id = {5c546e1a-664d-3c49-b780-ca530f662b55},\n created = {2020-12-17T05:29:55.706Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:55.706Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Vandevenne2014},\n source_type = {ARTICLE},\n notes = {cited By 3},\n private_publication = {false},\n abstract = {The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH &amp; Co. KGaA, Weinheim.},\n bibtype = {article},\n author = {Vandevenne, Marylène and O&#x27;Connell, Mitchell R. and Helder, Stephanie and Shepherd, Nicholas E. and Matthews, Jacqueline M. and Kwan, Ann H. and Segal, David J. and Mackay, Joel P.},\n doi = {10.1002/anie.201402980},\n journal = {Angewandte Chemie International Edition},\n number = {30}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://link.springer.com/10.1007/s12104-013-9470-x\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014', 'http://link.springer.com/10.1007/s12104-013-9470-x', event);\">\n    Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Joseph,  S.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H., Y.</a>; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Cubeddu,  L.;  and Matthews,  J., M.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 8(1): 141-144. 4 2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://link.springer.com/10.1007/s12104-013-9470-x\"\n     onclick=\"log_download('joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014', 'http://link.springer.com/10.1007/s12104-013-9470-x', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1 [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-013-9470-x\"\n     onclick=\"log_download('joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014', 'http://doi.org/10.1007/s12104-013-9470-x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1},\n type = {article},\n year = {2014},\n keywords = {Breast cancer,DEAF-1,Embryonic development,LMO4,Transcriptional complex},\n pages = {141-144},\n volume = {8},\n websites = {http://link.springer.com/10.1007/s12104-013-9470-x},\n month = {4},\n day = {16},\n id = {70636852-15f1-3af2-95ff-10b3cc12cad6},\n created = {2020-12-17T05:29:57.300Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:57.300Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Joseph2014a},\n source_type = {ARTICLE},\n notes = {cited By 2},\n private_publication = {false},\n abstract = {The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.},\n bibtype = {article},\n author = {Joseph, Soumya and Kwan, Ann H. Y. and Mackay, Joel P. and Cubeddu, Liza and Matthews, Jacqueline M.},\n doi = {10.1007/s12104-013-9470-x},\n journal = {Biomolecular NMR Assignments},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014', 'https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989', event);\">\n    Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Blythe,  A.; Gunasekara,  S.; Walshe,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Hartzog,  G., A.;  and Vrielink,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Expression and Purification</i>, 100: 54-60. 8 2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989\"\n     onclick=\"log_download('blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014', 'https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5 [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.pep.2014.05.005\"\n     onclick=\"log_download('blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014', 'http://doi.org/10.1016/j.pep.2014.05.005', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5},\n type = {article},\n year = {2014},\n keywords = {Expression and purification,KOW domain,Spt4/5,Ubiquitin tag,pHUE},\n pages = {54-60},\n volume = {100},\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S1046592814000989},\n month = {8},\n id = {c8758ecd-f201-3acf-9f44-56e1000a4f6a},\n created = {2020-12-17T05:29:57.964Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:57.964Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Blythe2014},\n source_type = {ARTICLE},\n notes = {cited By 1},\n private_publication = {false},\n abstract = {Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5&#x27;s mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.},\n bibtype = {article},\n author = {Blythe, Amanda and Gunasekara, Sanjika and Walshe, James and Mackay, Joel P. and Hartzog, Grant A. and Vrielink, Alice},\n doi = {10.1016/j.pep.2014.05.005},\n journal = {Protein Expression and Purification}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"clifton-westman-thong-oconnell-webster-shepherd-quinlan-crossley-etal-theidentificationandstructureofannterminalprdomainshowthatfog1isamemberoftheprdmfamilyofproteins-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The identification and structure of an N-terminal PR domain show that FOG1 is a member of the PRDM family of proteins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Clifton,  M.; Westman,  B.; Thong,  S.; O'Connell,  M.; Webster,  M.; Shepherd,  N.; Quinlan,  K.; Crossley,  M.; Blobel,  G.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>PLoS ONE</i>, 9(8).  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1371/journal.pone.0106011\"\n     onclick=\"log_download('clifton-westman-thong-oconnell-webster-shepherd-quinlan-crossley-etal-theidentificationandstructureofannterminalprdomainshowthatfog1isamemberoftheprdmfamilyofproteins-2014', 'http://doi.org/10.1371/journal.pone.0106011', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/clifton-westman-thong-oconnell-webster-shepherd-quinlan-crossley-etal-theidentificationandstructureofannterminalprdomainshowthatfog1isamemberoftheprdmfamilyofproteins-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('clifton-westman-thong-oconnell-webster-shepherd-quinlan-crossley-etal-theidentificationandstructureofannterminalprdomainshowthatfog1isamemberoftheprdmfamilyofproteins-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The identification and structure of an N-terminal PR domain show that FOG1 is a member of the PRDM family of proteins},\n type = {article},\n year = {2014},\n volume = {9},\n id = {d4bd4375-43f0-3b26-8cf7-d8fffdd6c864},\n created = {2023-01-10T01:44:35.718Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:35.718Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {FOG1 is a transcriptional regulator that acts in concert with the hematopoietic master regulator GATA1 to coordinate the differentiation of platelets and erythrocytes. Despite considerable effort, however, the mechanisms through which FOG1 regulates gene expression are only partially understood. Here we report the discovery of a previously unrecognized domain in FOG1: a PR (PRD-BF1 and RIZ) domain that is distantly related in sequence to the SET domains that are found in many histone methyltransferases. We have used NMR spectroscopy to determine the solution structure of this domain, revealing that the domain shares close structural similarity with SET domains. Titration with S-adenosyl-L-homocysteine, the cofactor product synonymous with SET domain methyltransferase activity, indicated that the FOG PR domain is not, however, likely to function as a methyltransferase in the same fashion. We also sought to define the function of this domain using both pulldown experiments and gel shift assays. However, neither pulldowns from mammalian nuclear extracts nor yeast two-hybrid assays reproducibly revealed binding partners, and we were unable to detect nucleic-acid-binding activity in this domain using our high-diversity Pentaprobe oligonucleotides. Overall, our data demonstrate that FOG1 is a member of the PRDM (PR domain containing proteins, with zinc fingers) family of transcriptional regulators. The function of many PR domains, however, remains somewhat enigmatic for the time being. © 2014 Clifton et al.},\n bibtype = {article},\n author = {Clifton, M.K. and Westman, B.J. and Thong, S.Y. and O&#x27;Connell, M.R. and Webster, M.W. and Shepherd, N.E. and Quinlan, K.G. and Crossley, M. and Blobel, G.A. and Mackay, J.P.},\n doi = {10.1371/journal.pone.0106011},\n journal = {PLoS ONE},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  FOG1 is a transcriptional regulator that acts in concert with the hematopoietic master regulator GATA1 to coordinate the differentiation of platelets and erythrocytes. Despite considerable effort, however, the mechanisms through which FOG1 regulates gene expression are only partially understood. Here we report the discovery of a previously unrecognized domain in FOG1: a PR (PRD-BF1 and RIZ) domain that is distantly related in sequence to the SET domains that are found in many histone methyltransferases. We have used NMR spectroscopy to determine the solution structure of this domain, revealing that the domain shares close structural similarity with SET domains. Titration with S-adenosyl-L-homocysteine, the cofactor product synonymous with SET domain methyltransferase activity, indicated that the FOG PR domain is not, however, likely to function as a methyltransferase in the same fashion. We also sought to define the function of this domain using both pulldown experiments and gel shift assays. However, neither pulldowns from mammalian nuclear extracts nor yeast two-hybrid assays reproducibly revealed binding partners, and we were unable to detect nucleic-acid-binding activity in this domain using our high-diversity Pentaprobe oligonucleotides. Overall, our data demonstrate that FOG1 is a member of the PRDM (PR domain containing proteins, with zinc fingers) family of transcriptional regulators. The function of many PR domains, however, remains somewhat enigmatic for the time being. © 2014 Clifton et al.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"cordina-liew-potluri-curmi-fajer-logan-mackay-brown-ca2inducedprenmrchangesinthetroponincomplexrevealthepossessivenatureofthecardiacisoformforitsregulatoryswitch-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Ca2+-induced PRE-NMR changes in the troponin complex reveal the possessive nature of the cardiac isoform for its regulatory switch.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Cordina,  N.; Liew,  C.; Potluri,  P.; Curmi,  P.; Fajer,  P.; Logan,  T.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Brown,  L.\n</span>\n\n  \n\n</span>\n\n  <i>PLoS ONE</i>, 9(11).  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1371/journal.pone.0112976\"\n     onclick=\"log_download('cordina-liew-potluri-curmi-fajer-logan-mackay-brown-ca2inducedprenmrchangesinthetroponincomplexrevealthepossessivenatureofthecardiacisoformforitsregulatoryswitch-2014', 'http://doi.org/10.1371/journal.pone.0112976', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/cordina-liew-potluri-curmi-fajer-logan-mackay-brown-ca2inducedprenmrchangesinthetroponincomplexrevealthepossessivenatureofthecardiacisoformforitsregulatoryswitch-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('cordina-liew-potluri-curmi-fajer-logan-mackay-brown-ca2inducedprenmrchangesinthetroponincomplexrevealthepossessivenatureofthecardiacisoformforitsregulatoryswitch-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Ca2+-induced PRE-NMR changes in the troponin complex reveal the possessive nature of the cardiac isoform for its regulatory switch},\n type = {article},\n year = {2014},\n volume = {9},\n id = {c3b7b413-fdec-325f-9bcd-b242670e45a7},\n created = {2023-01-10T01:44:36.628Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:36.628Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The interaction between myosin and actin in cardiac muscle, modulated by the calcium (Ca2+) sensor Troponin complex (Tn), is a complex process which is yet to be fully resolved at the molecular level. Our understanding of how the binding of Ca2+ triggers conformational changes within Tn that are subsequently propagated through the contractile apparatus to initiate muscle activation is hampered by a lack of an atomic structure for the Ca2+-free state of the cardiac isoform. We have used paramagnetic relaxation enhancement (PRE)-NMR to obtain a description of the Ca2+-free state of cardiac Tn by describing the movement of key regions of the troponin I (cTnI) subunit upon the release of Ca2+ from Troponin C (cTnC). Site-directed spin-labeling was used to position paramagnetic spin labels in cTnI and the changes in the interaction between cTnI and cTnC subunits were then mapped by PRE-NMR. The functionally important regions of cTnI targeted in this study included the cTnC-binding N-region (cTnI57), the inhibitory region (cTnI143), and two sites on the regulatory switch region (cTnI151 and cTnI159). Comparison of 1H-15N-TROSY spectra of Ca2+-bound and free states for the spin labeled cTnCcTnI binary constructs demonstrated the release and modest movement of the cTnI switch region (,10 A ) away from the hydrophobic N-lobe of troponin C (cTnC) upon the removal of Ca2+. Our data supports a model where the non-bound regulatory switch region of cTnI is highly flexible in the absence of Ca2+ but remains in close vicinity to cTnC. We speculate that the close proximity of TnI to TnC in the cardiac complex is favourable for increasing the frequency of collisions between the N-lobe of cTnC and the regulatory switch region, counterbalancing the reduction in collision probability that results from the incomplete opening of the N-lobe of TnC that is unique to the cardiac isoform.},\n bibtype = {article},\n author = {Cordina, N.M. and Liew, C.K. and Potluri, P.R. and Curmi, P.M. and Fajer, P.G. and Logan, T.M. and Mackay, J.P. and Brown, L.J.},\n doi = {10.1371/journal.pone.0112976},\n journal = {PLoS ONE},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The interaction between myosin and actin in cardiac muscle, modulated by the calcium (Ca2+) sensor Troponin complex (Tn), is a complex process which is yet to be fully resolved at the molecular level. Our understanding of how the binding of Ca2+ triggers conformational changes within Tn that are subsequently propagated through the contractile apparatus to initiate muscle activation is hampered by a lack of an atomic structure for the Ca2+-free state of the cardiac isoform. We have used paramagnetic relaxation enhancement (PRE)-NMR to obtain a description of the Ca2+-free state of cardiac Tn by describing the movement of key regions of the troponin I (cTnI) subunit upon the release of Ca2+ from Troponin C (cTnC). Site-directed spin-labeling was used to position paramagnetic spin labels in cTnI and the changes in the interaction between cTnI and cTnC subunits were then mapped by PRE-NMR. The functionally important regions of cTnI targeted in this study included the cTnC-binding N-region (cTnI57), the inhibitory region (cTnI143), and two sites on the regulatory switch region (cTnI151 and cTnI159). Comparison of 1H-15N-TROSY spectra of Ca2+-bound and free states for the spin labeled cTnCcTnI binary constructs demonstrated the release and modest movement of the cTnI switch region (,10 A ) away from the hydrophobic N-lobe of troponin C (cTnC) upon the removal of Ca2+. Our data supports a model where the non-bound regulatory switch region of cTnI is highly flexible in the absence of Ca2+ but remains in close vicinity to cTnC. We speculate that the close proximity of TnI to TnC in the cardiac complex is favourable for increasing the frequency of collisions between the N-lobe of cTnC and the regulatory switch region, counterbalancing the reduction in collision probability that results from the incomplete opening of the N-lobe of TnC that is unique to the cardiac isoform.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Engineering specificity changes on a RanBP2 zinc finger that binds single-stranded RNA.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Vandevenne,  M.; O'Connell,  M.; Helder,  S.; Shepherd,  N.; Matthews,  J.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Segal,  D.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Angewandte Chemie - International Edition</i>, 53(30): 7848-7852.  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/anie.201402980\"\n     onclick=\"log_download('vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014', 'http://doi.org/10.1002/anie.201402980', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('vandevenne-oconnell-helder-shepherd-matthews-kwan-segal-mackay-engineeringspecificitychangesonaranbp2zincfingerthatbindssinglestrandedrna-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Engineering specificity changes on a RanBP2 zinc finger that binds single-stranded RNA},\n type = {article},\n year = {2014},\n pages = {7848-7852},\n volume = {53},\n id = {efff0e09-c7d0-32a8-9607-835d587fd71a},\n created = {2023-01-10T01:44:37.599Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:37.599Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH &amp; Co. KGaA, Weinheim.},\n bibtype = {article},\n author = {Vandevenne, M. and O&#x27;Connell, M.R. and Helder, S. and Shepherd, N.E. and Matthews, J.M. and Kwan, A.H. and Segal, D.J. and Mackay, J.P.},\n doi = {10.1002/anie.201402980},\n journal = {Angewandte Chemie - International Edition},\n number = {30}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence. Variation on a theme: A combinatorial library of RanBP2-type zinc finger (ZF) domains has been engineered in an effort to select variants with distinct RNA-binding preferences. One variant was shown to successfully discriminate the sequence GCC over GGU and AAA, but only in the context of a three-ZF polypeptide. This study provides proof of principle that the specificity of RNA-binding modules based on ZF domains can be successfully altered. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"joseph-kwan-stokes-mackay-cubeddu-matthews-thestructureofanlimonlyprotein4lmo4anddeformedepidermalautoregulatoryfactor1deaf1complexrevealsacommonmodeofbindingtolmo4-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The structure of an LIM-Only protein 4 (LMO4) and Deformed Epidermal Autoregulatory Factor-1 (DEAF1) complex reveals a common mode of binding to LMO4.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Joseph,  S.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Stokes,  P.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Cubeddu,  L.;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>PLoS ONE</i>, 9(10).  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1371/journal.pone.0109108\"\n     onclick=\"log_download('joseph-kwan-stokes-mackay-cubeddu-matthews-thestructureofanlimonlyprotein4lmo4anddeformedepidermalautoregulatoryfactor1deaf1complexrevealsacommonmodeofbindingtolmo4-2014', 'http://doi.org/10.1371/journal.pone.0109108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/joseph-kwan-stokes-mackay-cubeddu-matthews-thestructureofanlimonlyprotein4lmo4anddeformedepidermalautoregulatoryfactor1deaf1complexrevealsacommonmodeofbindingtolmo4-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('joseph-kwan-stokes-mackay-cubeddu-matthews-thestructureofanlimonlyprotein4lmo4anddeformedepidermalautoregulatoryfactor1deaf1complexrevealsacommonmodeofbindingtolmo4-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The structure of an LIM-Only protein 4 (LMO4) and Deformed Epidermal Autoregulatory Factor-1 (DEAF1) complex reveals a common mode of binding to LMO4},\n type = {article},\n year = {2014},\n volume = {9},\n id = {5973cfa4-e9ae-3e89-b66f-cf09132c9ab9},\n created = {2023-01-10T01:44:38.507Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:38.507Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {LIM-domain only protein 4 (LMO4) is a widely expressed protein with important roles in embryonic development and breast cancer. It has been reported to bind many partners, including the transcription factor Deformed epidermal autoregulatory factor-1 (DEAF1), with which LMO4 shares many biological parallels. We used yeast two-hybrid assays to show that DEAF1 binds both LIM domains of LMO4 and that DEAF1 binds the same face on LMO4 as two other LMO4-binding partners, namely LIM domain binding protein 1 (LDB1) and C-terminal binding protein interacting protein (CtIP/RBBP8). Mutagenic screening analysed by the same method, indicates that the key residues in the interaction lie in LMO4LIM2and the N-terminal half of the LMO4-binding domain in DEAF1. We generated a stable LMO4LIM2-DEAF1 complex and determined the solution structure of that complex. Although the LMO4-binding domain from DEAF1 is intrinsically disordered, it becomes structured on binding. The structure confirms that LDB1, CtIP and DEAF1 all bind to the same face on LMO4. LMO4 appears to form a hub in protein-protein interaction networks, linking numerous pathways within cells. Competitive binding for LMO4 therefore most likely provides a level of regulation between those different pathways.},\n bibtype = {article},\n author = {Joseph, S. and Kwan, A.H. and Stokes, P.H. and Mackay, J.P. and Cubeddu, L. and Matthews, J.M.},\n doi = {10.1371/journal.pone.0109108},\n journal = {PLoS ONE},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  LIM-domain only protein 4 (LMO4) is a widely expressed protein with important roles in embryonic development and breast cancer. It has been reported to bind many partners, including the transcription factor Deformed epidermal autoregulatory factor-1 (DEAF1), with which LMO4 shares many biological parallels. We used yeast two-hybrid assays to show that DEAF1 binds both LIM domains of LMO4 and that DEAF1 binds the same face on LMO4 as two other LMO4-binding partners, namely LIM domain binding protein 1 (LDB1) and C-terminal binding protein interacting protein (CtIP/RBBP8). Mutagenic screening analysed by the same method, indicates that the key residues in the interaction lie in LMO4LIM2and the N-terminal half of the LMO4-binding domain in DEAF1. We generated a stable LMO4LIM2-DEAF1 complex and determined the solution structure of that complex. Although the LMO4-binding domain from DEAF1 is intrinsically disordered, it becomes structured on binding. The structure confirms that LDB1, CtIP and DEAF1 all bind to the same face on LMO4. LMO4 appears to form a hub in protein-protein interaction networks, linking numerous pathways within cells. Competitive binding for LMO4 therefore most likely provides a level of regulation between those different pathways.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"alqarni-murthy-zhang-przewloka-silva-watson-lejon-pei-etal-insightintothearchitectureofthenurdcomplexstructureoftherbap48mta1subcomplex-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Insight into the architecture of the NuRD complex: Structure of the RbAp48-MTA1 subcomplex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Alqarni,  S.; Murthy,  A.; Zhang,  W.; Przewloka,  M.; Silva,  A.; Watson,  A.; Lejon,  S.; Pei,  X.; Smits,  A.; Kloet,  S.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Laue,  E.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 289(32): 21844-21855.  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M114.558940\"\n     onclick=\"log_download('alqarni-murthy-zhang-przewloka-silva-watson-lejon-pei-etal-insightintothearchitectureofthenurdcomplexstructureoftherbap48mta1subcomplex-2014', 'http://doi.org/10.1074/jbc.M114.558940', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/alqarni-murthy-zhang-przewloka-silva-watson-lejon-pei-etal-insightintothearchitectureofthenurdcomplexstructureoftherbap48mta1subcomplex-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('alqarni-murthy-zhang-przewloka-silva-watson-lejon-pei-etal-insightintothearchitectureofthenurdcomplexstructureoftherbap48mta1subcomplex-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Insight into the architecture of the NuRD complex: Structure of the RbAp48-MTA1 subcomplex},\n type = {article},\n year = {2014},\n pages = {21844-21855},\n volume = {289},\n id = {a0afaf4d-99c7-3fea-9a0a-a55fcb967c4a},\n created = {2023-01-10T01:44:39.545Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:39.545Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background: The NuRD complex controls gene expression through altering chromatin structure. Results: The MTA1-RbAp48 structure shows how the RbAp46/p48 histone chaperones are recruited to NuRD. Conclusion: The MTA subunits act as scaffolds for NuRD complex assembly. Significance: The MTA/RbAp48 interaction prevents binding of histone H4, which is crucial for understanding the role of the RbAp46/p48 chaperones in the complex. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Alqarni, S.S.M. and Murthy, A. and Zhang, W. and Przewloka, M.R. and Silva, A.P.G. and Watson, A.A. and Lejon, S. and Pei, X.Y. and Smits, A.H. and Kloet, S.L. and Mackay, J.P. and Laue, E.D.},\n doi = {10.1074/jbc.M114.558940},\n journal = {Journal of Biological Chemistry},\n number = {32}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: The NuRD complex controls gene expression through altering chromatin structure. Results: The MTA1-RbAp48 structure shows how the RbAp46/p48 histone chaperones are recruited to NuRD. Conclusion: The MTA subunits act as scaffolds for NuRD complex assembly. Significance: The MTA/RbAp48 interaction prevents binding of histone H4, which is crucial for understanding the role of the RbAp46/p48 chaperones in the complex. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"dickson-kumar-jacques-malmirchegini-spirig-mackay-clubb-guss-etal-structureofthehemoglobinisdhcomplexrevealsthemolecularbasisofironcapturebystaphylococcusaureus-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structure of the hemoglobin-isdh complex reveals the molecular basis of iron capture by staphylococcus aureus.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dickson,  C.; Kumar,  K.; Jacques,  D.; Malmirchegini,  G.; Spirig,  T.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Clubb,  R.; Guss,  J.;  and Gell,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 289(10): 6728-6738.  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M113.545566\"\n     onclick=\"log_download('dickson-kumar-jacques-malmirchegini-spirig-mackay-clubb-guss-etal-structureofthehemoglobinisdhcomplexrevealsthemolecularbasisofironcapturebystaphylococcusaureus-2014', 'http://doi.org/10.1074/jbc.M113.545566', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dickson-kumar-jacques-malmirchegini-spirig-mackay-clubb-guss-etal-structureofthehemoglobinisdhcomplexrevealsthemolecularbasisofironcapturebystaphylococcusaureus-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dickson-kumar-jacques-malmirchegini-spirig-mackay-clubb-guss-etal-structureofthehemoglobinisdhcomplexrevealsthemolecularbasisofironcapturebystaphylococcusaureus-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structure of the hemoglobin-isdh complex reveals the molecular basis of iron capture by staphylococcus aureus},\n type = {article},\n year = {2014},\n pages = {6728-6738},\n volume = {289},\n id = {8082b276-9693-3b97-b289-93ddb85c7c38},\n created = {2023-01-10T01:44:40.488Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:40.488Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background: IsdB and IsdH proteins from Staphylococcus aureus strip heme iron from human hemoglobin. Results: The IsdH·hemoglobin complex shows how globin-binding and heme-binding NEAT domains of IsdH cooperate to remove heme from both chains of hemoglobin. Conclusion: The supradomain architecture of IsdH confers activity by precisely positioning the heme acceptor domain. Significance: Multiple IsdH·hemoglobin interfaces may be targets for new antibiotics. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Dickson, C.F. and Kumar, K.K. and Jacques, D.A. and Malmirchegini, G.R. and Spirig, T. and Mackay, J.P. and Clubb, R.T. and Guss, J.M. and Gell, D.A.},\n doi = {10.1074/jbc.M113.545566},\n journal = {Journal of Biological Chemistry},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: IsdB and IsdH proteins from Staphylococcus aureus strip heme iron from human hemoglobin. Results: The IsdH·hemoglobin complex shows how globin-binding and heme-binding NEAT domains of IsdH cooperate to remove heme from both chains of hemoglobin. Conclusion: The supradomain architecture of IsdH confers activity by precisely positioning the heme acceptor domain. Significance: Multiple IsdH·hemoglobin interfaces may be targets for new antibiotics. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Blythe,  A.; Gunasekara,  S.; Walshe,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Hartzog,  G.;  and Vrielink,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Expression and Purification</i>, 100: 54-60.  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.pep.2014.05.005\"\n     onclick=\"log_download('blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014', 'http://doi.org/10.1016/j.pep.2014.05.005', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('blythe-gunasekara-walshe-mackay-hartzog-vrielink-ubiquitinfusionconstructsallowtheexpressionandpurificationofmultikowdomaincomplexesofthesaccharomycescerevisiaetranscriptionelongationfactorspt45-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5},\n type = {article},\n year = {2014},\n pages = {54-60},\n volume = {100},\n id = {79bfac53-430c-3f33-98e9-37db63509339},\n created = {2023-01-10T01:44:41.424Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:41.424Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5&#x27;s mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.},\n bibtype = {article},\n author = {Blythe, A. and Gunasekara, S. and Walshe, J. and Mackay, J.P. and Hartzog, G.A. and Vrielink, A.},\n doi = {10.1016/j.pep.2014.05.005},\n journal = {Protein Expression and Purification}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. © 2014 Elsevier Inc. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Joseph,  S.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Cubeddu,  L.;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 8(1): 141-144.  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-013-9470-x\"\n     onclick=\"log_download('joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014', 'http://doi.org/10.1007/s12104-013-9470-x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('joseph-kwan-mackay-cubeddu-matthews-backboneandsidechainassignmentsofatetheredcomplexbetweenlmo4anddeaf1-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Backbone and side-chain assignments of a tethered complex between LMO4 and DEAF-1},\n type = {article},\n year = {2014},\n pages = {141-144},\n volume = {8},\n id = {a64344fa-3b32-37f0-b1e1-f7b7ad8cbfd8},\n created = {2023-01-10T01:44:42.330Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:42.330Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.},\n bibtype = {article},\n author = {Joseph, S. and Kwan, A.H.Y. and Mackay, J.P. and Cubeddu, L. and Matthews, J.M.},\n doi = {10.1007/s12104-013-9470-x},\n journal = {Biomolecular NMR Assignments},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The transcriptional regulator LMO4 and the transcription factor DEAF-1 are both essential for brain and skeletal development. They are also implicated in human breast cancers; overexpression of LMO4 is an indicator of poor prognosis, and overexpression of DEAF-1 promotes epithelial breast cell proliferation. We have generated a stable LMO4-DEAF-1 complex comprising the C-terminal LIM domain of LMO4 and an intrinsically disordered LMO4-interaction domain from DEAF-1 tethered by a glycine/serine linker. Here we report the 1H, 15N and 13C assignments of this construct. Analysis of the assignments indicates the presence of structure in the DEAF-1 part of the complex supporting the presence of a physical interaction between the two proteins. © 2013 Springer Science+Business Media Dordrecht.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"thom-traxler-khandros-nickas-zhou-lazarus-silva-prabhu-etal-trim58degradesdyneinandregulatesterminalerythropoiesis-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Trim58 Degrades Dynein and Regulates Terminal Erythropoiesis.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Thom,  C.; Traxler,  E.; Khandros,  E.; Nickas,  J.; Zhou,  O.; Lazarus,  J.; Silva,  A.; Prabhu,  D.; Yao,  Y.; Aribeana,  C.; Holzbaur,  E.;  and Weiss,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Developmental Cell</i>, 30(6): 688-700.  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.devcel.2014.07.021\"\n     onclick=\"log_download('thom-traxler-khandros-nickas-zhou-lazarus-silva-prabhu-etal-trim58degradesdyneinandregulatesterminalerythropoiesis-2014', 'http://doi.org/10.1016/j.devcel.2014.07.021', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/thom-traxler-khandros-nickas-zhou-lazarus-silva-prabhu-etal-trim58degradesdyneinandregulatesterminalerythropoiesis-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('thom-traxler-khandros-nickas-zhou-lazarus-silva-prabhu-etal-trim58degradesdyneinandregulatesterminalerythropoiesis-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Trim58 Degrades Dynein and Regulates Terminal Erythropoiesis},\n type = {article},\n year = {2014},\n pages = {688-700},\n volume = {30},\n id = {c1ccb790-7e19-39e4-aaad-2c8dd2dc11ef},\n created = {2023-01-10T01:44:43.258Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:43.258Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {TRIM58 is an E3 ubiquitin ligase superfamily member implicated by genome-wide association studies to regulate human erythrocyte traits. Here, we show that Trim58 expression is induced during late erythropoiesis and that its depletion by small hairpin RNAs (shRNAs) inhibits the maturation of late-stage nucleated erythroblasts to anucleate reticulocytes. Imaging flow cytometry studies demonstrate that Trim58 regulates polarization and/or extrusion of erythroblast nuclei. Invitro, Trim58 directly binds and ubiquitinates the intermediate chain of the microtubule motor dynein. In cells, Trim58 stimulates proteasome-dependent degradation of the dynein holoprotein complex. During erythropoiesis, Trim58 expression, dynein loss, and enucleation occur concomitantly, and all are inhibited by Trim58 shRNAs. Dynein regulates nuclear positioning and microtubule organization, both of which undergo dramatic changes during erythroblast enucleation. Thus, we propose that Trim58 promotes this processby eliminating dynein. Our findings identify an erythroid-specific regulator of enucleation and elucidate a previously unrecognized mechanism for controlling dynein activity.},\n bibtype = {article},\n author = {Thom, C.S. and Traxler, E.A. and Khandros, E. and Nickas, J.M. and Zhou, O.Y. and Lazarus, J.E. and Silva, A.P.G. and Prabhu, D. and Yao, Y. and Aribeana, C. and Holzbaur, E.L.F. and Weiss, M.J.},\n doi = {10.1016/j.devcel.2014.07.021},\n journal = {Developmental Cell},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  TRIM58 is an E3 ubiquitin ligase superfamily member implicated by genome-wide association studies to regulate human erythrocyte traits. Here, we show that Trim58 expression is induced during late erythropoiesis and that its depletion by small hairpin RNAs (shRNAs) inhibits the maturation of late-stage nucleated erythroblasts to anucleate reticulocytes. Imaging flow cytometry studies demonstrate that Trim58 regulates polarization and/or extrusion of erythroblast nuclei. Invitro, Trim58 directly binds and ubiquitinates the intermediate chain of the microtubule motor dynein. In cells, Trim58 stimulates proteasome-dependent degradation of the dynein holoprotein complex. During erythropoiesis, Trim58 expression, dynein loss, and enucleation occur concomitantly, and all are inhibited by Trim58 shRNAs. Dynein regulates nuclear positioning and microtubule organization, both of which undergo dramatic changes during erythroblast enucleation. Thus, we propose that Trim58 promotes this processby eliminating dynein. Our findings identify an erythroid-specific regulator of enucleation and elucidate a previously unrecognized mechanism for controlling dynein activity.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-transcriptionfactorseeksdnacognatesitepreferred-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Transcription factor seeks DNA - Cognate site preferred.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 426(7): 1370-1372.  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2013.12.010\"\n     onclick=\"log_download('mackay-transcriptionfactorseeksdnacognatesitepreferred-2014', 'http://doi.org/10.1016/j.jmb.2013.12.010', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-transcriptionfactorseeksdnacognatesitepreferred-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-transcriptionfactorseeksdnacognatesitepreferred-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Transcription factor seeks DNA - Cognate site preferred},\n type = {article},\n year = {2014},\n pages = {1370-1372},\n volume = {426},\n id = {1e075f30-6417-39c7-8b24-b13d5c28f504},\n created = {2023-01-10T01:44:44.178Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:44.178Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Mackay, J.},\n doi = {10.1016/j.jmb.2013.12.010},\n journal = {Journal of Molecular Biology},\n number = {7}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"silva-kwan-mackay-sup1suphsup13supcandsup15supnresonanceassignmentsofannterminaldomainofchd4-2014\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N resonance assignments of an N-terminal domain of CHD4.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Silva,  A.; Kwan,  A.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 8(1): 137-139.  2014.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-013-9469-3\"\n     onclick=\"log_download('silva-kwan-mackay-sup1suphsup13supcandsup15supnresonanceassignmentsofannterminaldomainofchd4-2014', 'http://doi.org/10.1007/s12104-013-9469-3', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/silva-kwan-mackay-sup1suphsup13supcandsup15supnresonanceassignmentsofannterminaldomainofchd4-2014\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('silva-kwan-mackay-sup1suphsup13supcandsup15supnresonanceassignmentsofannterminaldomainofchd4-2014')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {&lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;13&lt;/sup&gt;C and &lt;sup&gt;15&lt;/sup&gt;N resonance assignments of an N-terminal domain of CHD4},\n type = {article},\n year = {2014},\n pages = {137-139},\n volume = {8},\n id = {cd0244ce-1944-3196-a041-f999ed20f6c3},\n created = {2023-01-10T01:44:45.086Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:45.086Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.},\n bibtype = {article},\n author = {Silva, A.P.G. and Kwan, A.H. and Mackay, J.P.},\n doi = {10.1007/s12104-013-9469-3},\n journal = {Biomolecular NMR Assignments},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chromatin-remodeling proteins have a pivotal role in normal cell function and development, catalyzing conformational changes in DNA that ultimately result in changes in gene expression patterns. Chromodomain helicase DNA-binding protein 4 (CHD4), the defining subunit of the nucleosome remodeling and deacetylase (NuRD) complex, is a nucleosome-remodeling protein of the SNF2/ISWI2 family, members of which contain two chromo domains and an ATP-dependent helicase module. CHD3, CHD4 and CHD5 also contain two contiguous PHD domains and have an extended N-terminal region that has not previously been characterized. We have identified a stable domain in the N-terminal region of CHD4 and report here the backbone and side chain resonance assignments for this domain at pH 7.5 and 25 °C (BMRB No. 18906). © 2013 Springer Science+Business Media Dordrecht.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2013\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2013')\"\n      onkeypress=\"toggleGroup('group_2013')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2013</span>\n      <span class=\"bibbase_group_count\">\n        \n        (8)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"dickson-rich-davigdor-collins-lowry-mollan-khandros-olson-etal-hemoglobinstabilizingproteinahspperturbstheproximalhemepocketofoxyhemoglobinandweakenstheironoxygenbond-2013\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  α-Hemoglobin-stabilizing Protein (AHSP) perturbs the proximal heme pocket of oxy-α-hemoglobin and weakens the iron-oxygen bond.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dickson,  C.; Rich,  A.; D'Avigdor,  W.; Collins,  D.; Lowry,  J.; Mollan,  T.; Khandros,  E.; Olson,  J.; Weiss,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Lay,  P.;  and Gell,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 288(27): 19986-20001.  2013.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M112.437509\"\n     onclick=\"log_download('dickson-rich-davigdor-collins-lowry-mollan-khandros-olson-etal-hemoglobinstabilizingproteinahspperturbstheproximalhemepocketofoxyhemoglobinandweakenstheironoxygenbond-2013', 'http://doi.org/10.1074/jbc.M112.437509', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dickson-rich-davigdor-collins-lowry-mollan-khandros-olson-etal-hemoglobinstabilizingproteinahspperturbstheproximalhemepocketofoxyhemoglobinandweakenstheironoxygenbond-2013\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dickson-rich-davigdor-collins-lowry-mollan-khandros-olson-etal-hemoglobinstabilizingproteinahspperturbstheproximalhemepocketofoxyhemoglobinandweakenstheironoxygenbond-2013')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {α-Hemoglobin-stabilizing Protein (AHSP) perturbs the proximal heme pocket of oxy-α-hemoglobin and weakens the iron-oxygen bond},\n type = {article},\n year = {2013},\n pages = {19986-20001},\n volume = {288},\n id = {d5cb8d14-4ed7-37da-a53b-1987811758ed},\n created = {2023-01-10T01:44:46.024Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:46.024Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {α-Hemoglobin (αHb)-stabilizing protein (AHSP) is a molecular chaperone that assists hemoglobin assembly. AHSP induces changes in αHb heme coordination, but how these changes are facilitated by interactions at the αHb·AHSP interface is not well understood. To address this question we have used NMR, x-ray absorption spectroscopy, and ligand binding measurements to probe αHb conformational changes induced by AHSP binding.NMRchemical shift analyses of free CO-αHb and CO- αHb·AHSP indicated that the seven helical elements of the native αHb structure are retained and that the heme Fe(II) remains coordinated to the proximal His-87 side chain. However, chemical shift differences revealed alterations of the F, G, and H helices and the heme pocket of CO-αHb bound to AHSP. Comparisons of iron-ligand geometry using extended x-ray absorption fine structure spectroscopy showed that AHSP binding induces a small 0.03 Å lengthening of the Fe-O2 bond, explaining previous reports that AHSP decreases αHb O2 affinity roughly 4-fold and promotes autooxidation due primarily to a 3-4-fold increase in the rate ofO2 dissociation. Pro-30 mutations diminished NMR chemical shift changes in the proximal heme pocket, restored normal O2 dissociation rate and equilibrium constants, and reduced O2-αHb autooxidation rates. Thus, the contacts mediated by Pro-30 in wild-type AHSP promote αHb autooxidation by introducing strain into the proximal heme pocket. As a chaperone, AHSP facilitates rapid assembly of αHb into Hb when βHb is abundant but diverts αHb to a redox resistant holding state when βHb is limiting. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Dickson, C.F. and Rich, A.M. and D&#x27;Avigdor, W.M.H. and Collins, D.A.T. and Lowry, J.A. and Mollan, T.L. and Khandros, E. and Olson, J.S. and Weiss, M.J. and MacKay, J.P. and Lay, P.A. and Gell, D.A.},\n doi = {10.1074/jbc.M112.437509},\n journal = {Journal of Biological Chemistry},\n number = {27}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  α-Hemoglobin (αHb)-stabilizing protein (AHSP) is a molecular chaperone that assists hemoglobin assembly. AHSP induces changes in αHb heme coordination, but how these changes are facilitated by interactions at the αHb·AHSP interface is not well understood. To address this question we have used NMR, x-ray absorption spectroscopy, and ligand binding measurements to probe αHb conformational changes induced by AHSP binding.NMRchemical shift analyses of free CO-αHb and CO- αHb·AHSP indicated that the seven helical elements of the native αHb structure are retained and that the heme Fe(II) remains coordinated to the proximal His-87 side chain. However, chemical shift differences revealed alterations of the F, G, and H helices and the heme pocket of CO-αHb bound to AHSP. Comparisons of iron-ligand geometry using extended x-ray absorption fine structure spectroscopy showed that AHSP binding induces a small 0.03 Å lengthening of the Fe-O2 bond, explaining previous reports that AHSP decreases αHb O2 affinity roughly 4-fold and promotes autooxidation due primarily to a 3-4-fold increase in the rate ofO2 dissociation. Pro-30 mutations diminished NMR chemical shift changes in the proximal heme pocket, restored normal O2 dissociation rate and equilibrium constants, and reduced O2-αHb autooxidation rates. Thus, the contacts mediated by Pro-30 in wild-type AHSP promote αHb autooxidation by introducing strain into the proximal heme pocket. As a chaperone, AHSP facilitates rapid assembly of αHb into Hb when βHb is abundant but diverts αHb to a redox resistant holding state when βHb is limiting. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-segal-crossley-isthereatelltalerhfingerprintinzincfingersthatrecognizesmethylatedcpgdinucleotides-2013\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Is there a telltale RH fingerprint in zinc fingers that recognizes methylated CpG dinucleotides?.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Segal,  D.;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Trends in Biochemical Sciences</i>, 38(9): 421-422.  2013.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.tibs.2013.06.005\"\n     onclick=\"log_download('mackay-segal-crossley-isthereatelltalerhfingerprintinzincfingersthatrecognizesmethylatedcpgdinucleotides-2013', 'http://doi.org/10.1016/j.tibs.2013.06.005', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-segal-crossley-isthereatelltalerhfingerprintinzincfingersthatrecognizesmethylatedcpgdinucleotides-2013\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-segal-crossley-isthereatelltalerhfingerprintinzincfingersthatrecognizesmethylatedcpgdinucleotides-2013')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Is there a telltale RH fingerprint in zinc fingers that recognizes methylated CpG dinucleotides?},\n type = {article},\n year = {2013},\n pages = {421-422},\n volume = {38},\n id = {4f05c721-c533-3f78-93e4-e5649cccf00b},\n created = {2023-01-10T01:44:46.983Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:46.983Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Mackay, J.P. and Segal, D.J. and Crossley, M.},\n doi = {10.1016/j.tibs.2013.06.005},\n journal = {Trends in Biochemical Sciences},\n number = {9}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gamsjaeger-oconnell-cubeddu-shepherd-lowry-kwan-vandevenne-swanton-etal-astructuralanalysisofdnabindingbymyelintranscriptionfactor1doublezincfingers-2013\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A structural analysis of DNA binding by myelin transcription factor 1 double zinc fingers.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gamsjaeger,  R.; O'Connell,  M.; Cubeddu,  L.; Shepherd,  N.; Lowry,  J.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Vandevenne,  M.; Swanton,  M.; Matthews,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 288(49): 35180-35191.  2013.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M113.482075\"\n     onclick=\"log_download('gamsjaeger-oconnell-cubeddu-shepherd-lowry-kwan-vandevenne-swanton-etal-astructuralanalysisofdnabindingbymyelintranscriptionfactor1doublezincfingers-2013', 'http://doi.org/10.1074/jbc.M113.482075', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gamsjaeger-oconnell-cubeddu-shepherd-lowry-kwan-vandevenne-swanton-etal-astructuralanalysisofdnabindingbymyelintranscriptionfactor1doublezincfingers-2013\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gamsjaeger-oconnell-cubeddu-shepherd-lowry-kwan-vandevenne-swanton-etal-astructuralanalysisofdnabindingbymyelintranscriptionfactor1doublezincfingers-2013')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A structural analysis of DNA binding by myelin transcription factor 1 double zinc fingers},\n type = {article},\n year = {2013},\n pages = {35180-35191},\n volume = {288},\n id = {87086ec5-3591-3e63-826a-b7a3fde32906},\n created = {2023-01-10T01:44:47.925Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:47.925Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background: Myelin transcription factor 1 (MyT1) contains seven similar zinc finger domains that bind DNA specifically. Results: A three-dimensional structural model explains how a double zinc finger unit is able to recognize DNA. Conclusion: DNA-binding residues are conserved among all MyT1 zinc fingers, suggesting an identical DNA binding mode. Significance: Determination of the molecular details of DNA interaction will be crucial in understanding MyT1 function. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Gamsjaeger, R. and O&#x27;Connell, M.R. and Cubeddu, L. and Shepherd, N.E. and Lowry, J.A. and Kwan, A.H. and Vandevenne, M. and Swanton, M.K. and Matthews, J.M. and Mackay, J.P.},\n doi = {10.1074/jbc.M113.482075},\n journal = {Journal of Biological Chemistry},\n number = {49}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: Myelin transcription factor 1 (MyT1) contains seven similar zinc finger domains that bind DNA specifically. Results: A three-dimensional structural model explains how a double zinc finger unit is able to recognize DNA. Conclusion: DNA-binding residues are conserved among all MyT1 zinc fingers, suggesting an identical DNA binding mode. Significance: Determination of the molecular details of DNA interaction will be crucial in understanding MyT1 function. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"vandevenne-jacques-artuz-nguyen-kwan-segal-matthews-crossley-etal-newinsightsintodnarecognitionbyzincfingersrevealedbystructuralanalysisoftheoncoproteinznf217-2013\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  New insights into DNA recognition by zinc fingers revealed by structural analysis of the oncoprotein ZNF217.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Vandevenne,  M.; Jacques,  D.; Artuz,  C.; Nguyen,  C.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Segal,  D.; Matthews,  J.; Crossley,  M.; Guss,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 288(15): 10616-10627.  2013.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M112.441451\"\n     onclick=\"log_download('vandevenne-jacques-artuz-nguyen-kwan-segal-matthews-crossley-etal-newinsightsintodnarecognitionbyzincfingersrevealedbystructuralanalysisoftheoncoproteinznf217-2013', 'http://doi.org/10.1074/jbc.M112.441451', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/vandevenne-jacques-artuz-nguyen-kwan-segal-matthews-crossley-etal-newinsightsintodnarecognitionbyzincfingersrevealedbystructuralanalysisoftheoncoproteinznf217-2013\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('vandevenne-jacques-artuz-nguyen-kwan-segal-matthews-crossley-etal-newinsightsintodnarecognitionbyzincfingersrevealedbystructuralanalysisoftheoncoproteinznf217-2013')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {New insights into DNA recognition by zinc fingers revealed by structural analysis of the oncoprotein ZNF217},\n type = {article},\n year = {2013},\n pages = {10616-10627},\n volume = {288},\n id = {406b142f-54cb-305c-8eaa-3b8866da36f3},\n created = {2023-01-10T01:44:48.847Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:48.847Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background: Classical zinc finger proteins are extremely abundant and interact with DNA using a well defined recognition code. Results: We solved the structure of ZNF217 bound to its cognate DNA. Conclusion: ZNF217 presents a unique DNA interaction pattern including a new type of protein-DNA contact. Significance: This study deepens our understanding of DNA recognition by classical zinc fingers. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Vandevenne, M. and Jacques, D.A. and Artuz, C. and Nguyen, C.D. and Kwan, A.H.Y. and Segal, D.J. and Matthews, J.M. and Crossley, M. and Guss, J.M. and MacKay, J.P.},\n doi = {10.1074/jbc.M112.441451},\n journal = {Journal of Biological Chemistry},\n number = {15}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: Classical zinc finger proteins are extremely abundant and interact with DNA using a well defined recognition code. Results: We solved the structure of ZNF217 bound to its cognate DNA. Conclusion: ZNF217 presents a unique DNA interaction pattern including a new type of protein-DNA contact. Significance: This study deepens our understanding of DNA recognition by classical zinc fingers. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"cordina-liew-gell-fajer-mackay-brown-effectsofcalciumbindingandthehypertrophiccardiomyopathya8vmutationonthedynamicequilibriumbetweenclosedandopenconformationsoftheregulatoryndomainofisolatedcardiactroponinc-2013\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Effects of calcium binding and the hypertrophic cardiomyopathy A8V mutation on the dynamic equilibrium between closed and open conformations of the regulatory N-domain of isolated cardiac troponin C.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Cordina,  N.; Liew,  C.; Gell,  D.; Fajer,  P.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Brown,  L.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 52(11): 1950-1962.  2013.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi4000172\"\n     onclick=\"log_download('cordina-liew-gell-fajer-mackay-brown-effectsofcalciumbindingandthehypertrophiccardiomyopathya8vmutationonthedynamicequilibriumbetweenclosedandopenconformationsoftheregulatoryndomainofisolatedcardiactroponinc-2013', 'http://doi.org/10.1021/bi4000172', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/cordina-liew-gell-fajer-mackay-brown-effectsofcalciumbindingandthehypertrophiccardiomyopathya8vmutationonthedynamicequilibriumbetweenclosedandopenconformationsoftheregulatoryndomainofisolatedcardiactroponinc-2013\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('cordina-liew-gell-fajer-mackay-brown-effectsofcalciumbindingandthehypertrophiccardiomyopathya8vmutationonthedynamicequilibriumbetweenclosedandopenconformationsoftheregulatoryndomainofisolatedcardiactroponinc-2013')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Effects of calcium binding and the hypertrophic cardiomyopathy A8V mutation on the dynamic equilibrium between closed and open conformations of the regulatory N-domain of isolated cardiac troponin C},\n type = {article},\n year = {2013},\n pages = {1950-1962},\n volume = {52},\n id = {a6e23941-429e-3ffe-905a-69c9fa9461ee},\n created = {2023-01-10T01:44:49.760Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:49.760Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Troponin C (TnC) is the calcium-binding subunit of the troponin complex responsible for initiating striated muscle contraction in response to calcium influx. In the skeletal TnC isoform, calcium binding induces a structural change in the regulatory N-domain of TnC that involves a transition from a closed to open structural state and accompanying exposure of a large hydrophobic patch for troponin I (TnI) to subsequently bind. However, little is understood about how calcium primes the N-domain of the cardiac isoform (cTnC) for interaction with the TnI subunit as the open conformation of the regulatory domain of cTnC has been observed only in the presence of bound TnI. Here we use paramagnetic relaxation enhancement (PRE) to characterize the closed to open transition of isolated cTnC in solution, a process that cannot be observed by traditional nuclear magnetic resonance methods. Our PRE data from four spin-labeled monocysteine constructs of isolated cTnC reveal that calcium binding triggers movement of the N-domain helices toward an open state. Fitting of the PRE data to a closed to open transition model reveals the presence of a small population of cTnC molecules in the absence of calcium that possess an open conformation, the level of which increases substantially upon Ca2+ binding. These data support a model in which calcium binding creates a dynamic equilibrium between the closed and open structural states to prime cTnC for interaction with its target peptide. We also used PRE data to assess the structural effects of a familial hypertrophic cardiomyopathy point mutation located within the N-domain of cTnC (A8V). The PRE data show that the Ca2+ switch mechanism is perturbed by the A8V mutation, resulting in a more open N-domain conformation in both the apo and holo states. © 2013 American Chemical Society.},\n bibtype = {article},\n author = {Cordina, N.M. and Liew, C.K. and Gell, D.A. and Fajer, P.G. and MacKay, J.P. and Brown, L.J.},\n doi = {10.1021/bi4000172},\n journal = {Biochemistry},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Troponin C (TnC) is the calcium-binding subunit of the troponin complex responsible for initiating striated muscle contraction in response to calcium influx. In the skeletal TnC isoform, calcium binding induces a structural change in the regulatory N-domain of TnC that involves a transition from a closed to open structural state and accompanying exposure of a large hydrophobic patch for troponin I (TnI) to subsequently bind. However, little is understood about how calcium primes the N-domain of the cardiac isoform (cTnC) for interaction with the TnI subunit as the open conformation of the regulatory domain of cTnC has been observed only in the presence of bound TnI. Here we use paramagnetic relaxation enhancement (PRE) to characterize the closed to open transition of isolated cTnC in solution, a process that cannot be observed by traditional nuclear magnetic resonance methods. Our PRE data from four spin-labeled monocysteine constructs of isolated cTnC reveal that calcium binding triggers movement of the N-domain helices toward an open state. Fitting of the PRE data to a closed to open transition model reveals the presence of a small population of cTnC molecules in the absence of calcium that possess an open conformation, the level of which increases substantially upon Ca2+ binding. These data support a model in which calcium binding creates a dynamic equilibrium between the closed and open structural states to prime cTnC for interaction with its target peptide. We also used PRE data to assess the structural effects of a familial hypertrophic cardiomyopathy point mutation located within the N-domain of cTnC (A8V). The PRE data show that the Ca2+ switch mechanism is perturbed by the A8V mutation, resulting in a more open N-domain conformation in both the apo and holo states. © 2013 American Chemical Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"stokes-liew-kwan-foo-barker-djamirze-oreilly-visvader-etal-structuralbasisoftheinteractionofthebreastcanceroncogenelmo4withthetumoursuppressorctiprbbp8-2013\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural basis of the interaction of the breast cancer Oncogene LMO4 with the tumour suppressor CtIP/RBBP8.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Stokes,  P.; Liew,  C.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Foo,  P.; Barker,  H.; Djamirze,  A.; O'Reilly,  V.; Visvader,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 425(7): 1101-1110.  2013.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2013.01.017\"\n     onclick=\"log_download('stokes-liew-kwan-foo-barker-djamirze-oreilly-visvader-etal-structuralbasisoftheinteractionofthebreastcanceroncogenelmo4withthetumoursuppressorctiprbbp8-2013', 'http://doi.org/10.1016/j.jmb.2013.01.017', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/stokes-liew-kwan-foo-barker-djamirze-oreilly-visvader-etal-structuralbasisoftheinteractionofthebreastcanceroncogenelmo4withthetumoursuppressorctiprbbp8-2013\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('stokes-liew-kwan-foo-barker-djamirze-oreilly-visvader-etal-structuralbasisoftheinteractionofthebreastcanceroncogenelmo4withthetumoursuppressorctiprbbp8-2013')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural basis of the interaction of the breast cancer Oncogene LMO4 with the tumour suppressor CtIP/RBBP8},\n type = {article},\n year = {2013},\n pages = {1101-1110},\n volume = {425},\n id = {191b6ccd-b3f5-3f12-a76e-f0e33183952f},\n created = {2023-01-10T01:44:50.675Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:50.675Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {LIM-only protein 4 (LMO4) is strongly linked to the progression of breast cancer. Although the mechanisms underlying this phenomenon are not well understood, a role is emerging for LMO4 in regulation of the cell cycle. We determined the solution structure of LMO4 in complex with CtIP (C-terminal binding protein interacting protein)/RBBP8, a tumour suppressor protein that is involved in cell cycle progression, DNA repair and transcriptional regulation. Our data reveal that CtIP and the essential LMO cofactor LDB1 (LIM-domain binding protein 1) bind to the same face on LMO4 and cannot simultaneously bind to LMO4. We hypothesise that overexpression of LMO4 may disrupt some of the normal tumour suppressor activities of CtIP, thereby contributing to breast cancer progression. © 2013 Elsevier Ltd.},\n bibtype = {article},\n author = {Stokes, P.H. and Liew, C.W. and Kwan, A.H. and Foo, P. and Barker, H.E. and Djamirze, A. and O&#x27;Reilly, V. and Visvader, J.E. and Mackay, J.P. and Matthews, J.M.},\n doi = {10.1016/j.jmb.2013.01.017},\n journal = {Journal of Molecular Biology},\n number = {7}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  LIM-only protein 4 (LMO4) is strongly linked to the progression of breast cancer. Although the mechanisms underlying this phenomenon are not well understood, a role is emerging for LMO4 in regulation of the cell cycle. We determined the solution structure of LMO4 in complex with CtIP (C-terminal binding protein interacting protein)/RBBP8, a tumour suppressor protein that is involved in cell cycle progression, DNA repair and transcriptional regulation. Our data reveal that CtIP and the essential LMO cofactor LDB1 (LIM-domain binding protein 1) bind to the same face on LMO4 and cannot simultaneously bind to LMO4. We hypothesise that overexpression of LMO4 may disrupt some of the normal tumour suppressor activities of CtIP, thereby contributing to breast cancer progression. © 2013 Elsevier Ltd.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gamsjaeger-kariawasam-bang-touma-nguyen-matthews-cubeddu-mackay-semiquantitativeandquantitativeanalysisofproteindnainteractionsusingsteadystatemeasurementsinsurfaceplasmonresonancecompetitionexperiments-2013\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Semiquantitative and quantitative analysis of protein-DNA interactions using steady-state measurements in surface plasmon resonance competition experiments.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gamsjaeger,  R.; Kariawasam,  R.; Bang,  L.; Touma,  C.; Nguyen,  C.; Matthews,  J.; Cubeddu,  L.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Analytical Biochemistry</i>, 440(2): 178-185.  2013.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.ab.2013.04.030\"\n     onclick=\"log_download('gamsjaeger-kariawasam-bang-touma-nguyen-matthews-cubeddu-mackay-semiquantitativeandquantitativeanalysisofproteindnainteractionsusingsteadystatemeasurementsinsurfaceplasmonresonancecompetitionexperiments-2013', 'http://doi.org/10.1016/j.ab.2013.04.030', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gamsjaeger-kariawasam-bang-touma-nguyen-matthews-cubeddu-mackay-semiquantitativeandquantitativeanalysisofproteindnainteractionsusingsteadystatemeasurementsinsurfaceplasmonresonancecompetitionexperiments-2013\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gamsjaeger-kariawasam-bang-touma-nguyen-matthews-cubeddu-mackay-semiquantitativeandquantitativeanalysisofproteindnainteractionsusingsteadystatemeasurementsinsurfaceplasmonresonancecompetitionexperiments-2013')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Semiquantitative and quantitative analysis of protein-DNA interactions using steady-state measurements in surface plasmon resonance competition experiments},\n type = {article},\n year = {2013},\n pages = {178-185},\n volume = {440},\n id = {cb78304b-2ce1-32bf-ae2e-25e278ca3b2c},\n created = {2023-01-10T01:44:51.607Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:51.607Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {One method commonly used to characterize protein-DNA interactions is surface plasmon resonance (SPR). In a typical SPR experiment, chip-bound DNA is exposed to increasing concentrations of protein; the resulting binding data are used to calculate a dissociation constant for the interaction. However, in cases in which knowledge of the specificity of the interaction is required, a large set of DNA variants has to be tested; this is time consuming and costly, in part because of the requirement for multiple SPR chips. We have developed a new protocol that uses steady-state binding levels in SPR competition experiments to determine protein-binding dissociation constants for a set of DNA variants. This approach is rapid and straightforward and requires the use of only a single SPR chip. Additionally, in contrast to other methods, our approach does not require prior knowledge of parameters such as on or off rates, using an estimate of the wild-type interaction as the sole input. Utilizing relative steady-state responses, our protocol also allows for the rapid, reliable, and simultaneous determination of protein-binding dissociation constants of a large series of DNA mutants in a single experiment in a semiquantitative fashion. We compare our approach to existing methods, highlighting specific advantages as well as limitations. © 2013 Elsevier Inc. All rights reserved.},\n bibtype = {article},\n author = {Gamsjaeger, R. and Kariawasam, R. and Bang, L.H. and Touma, C. and Nguyen, C.D. and Matthews, J.M. and Cubeddu, L. and Mackay, J.P.},\n doi = {10.1016/j.ab.2013.04.030},\n journal = {Analytical Biochemistry},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  One method commonly used to characterize protein-DNA interactions is surface plasmon resonance (SPR). In a typical SPR experiment, chip-bound DNA is exposed to increasing concentrations of protein; the resulting binding data are used to calculate a dissociation constant for the interaction. However, in cases in which knowledge of the specificity of the interaction is required, a large set of DNA variants has to be tested; this is time consuming and costly, in part because of the requirement for multiple SPR chips. We have developed a new protocol that uses steady-state binding levels in SPR competition experiments to determine protein-binding dissociation constants for a set of DNA variants. This approach is rapid and straightforward and requires the use of only a single SPR chip. Additionally, in contrast to other methods, our approach does not require prior knowledge of parameters such as on or off rates, using an estimate of the wild-type interaction as the sole input. Utilizing relative steady-state responses, our protocol also allows for the rapid, reliable, and simultaneous determination of protein-binding dissociation constants of a large series of DNA mutants in a single experiment in a semiquantitative fashion. We compare our approach to existing methods, highlighting specific advantages as well as limitations. © 2013 Elsevier Inc. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"campbell-wilkinsonwhite-mackay-matthews-blobel-analysisofdiseasecausinggata1mutationsinmurinegenecomplementationsystems-2013\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Analysis of disease-causing GATA1 mutations in murine gene complementation systems.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Campbell,  A.; Wilkinson-White,  L.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Matthews,  J.;  and Blobel,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Blood</i>, 121(26): 5218-5227.  2013.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1182/blood-2013-03-488080\"\n     onclick=\"log_download('campbell-wilkinsonwhite-mackay-matthews-blobel-analysisofdiseasecausinggata1mutationsinmurinegenecomplementationsystems-2013', 'http://doi.org/10.1182/blood-2013-03-488080', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/campbell-wilkinsonwhite-mackay-matthews-blobel-analysisofdiseasecausinggata1mutationsinmurinegenecomplementationsystems-2013\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('campbell-wilkinsonwhite-mackay-matthews-blobel-analysisofdiseasecausinggata1mutationsinmurinegenecomplementationsystems-2013')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Analysis of disease-causing GATA1 mutations in murine gene complementation systems},\n type = {article},\n year = {2013},\n pages = {5218-5227},\n volume = {121},\n id = {ddfb0231-026d-32b5-a980-22e8754a9d8b},\n created = {2023-01-10T01:44:52.681Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:52.681Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Missense mutations in transcription factor GATA1 underlie a spectrum of congenital red blood cell and platelet disorders. We investigated how these alterations cause distinct clinical phenotypes by combining structural, biochemical, and genomic approaches with gene complementation systems that examine GATA1 function in biologically relevant cellular contexts. Substitutions that disrupt FOG1 cofactor binding impair both gene activation and repression and are associated with pronounced clinical phenotypes. Moreover, clinical severity correlates with the degree of FOG1 disruption. Surprisingly, 2 mutations shown to impair DNA binding of GATA1 in vitro did not measurably affect in vivo target gene occupancy. Rather, one of these disrupted binding to the TAL1 complex, implicating it in diseases caused by GATA1 mutations. Diminished TAL1 complex recruitment mainly impairs transcriptional activation and is linked to relatively mild disease. Notably, different substitutions at the same amino acid can selectively inhibit TAL1 complex or FOG1 binding, producing distinct cellular and clinical phenotypes. The structure-function relationships elucidated here were not predicted by prior in vitro or computational studies. Thus, our findings uncover novel disease mechanisms underlying GATA1 mutations and highlight the power of gene complementation assays for elucidating the molecular basis of genetic diseases.},\n bibtype = {article},\n author = {Campbell, A.E. and Wilkinson-White, L. and MacKay, J.P. and Matthews, J.M. and Blobel, G.A.},\n doi = {10.1182/blood-2013-03-488080},\n journal = {Blood},\n number = {26}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Missense mutations in transcription factor GATA1 underlie a spectrum of congenital red blood cell and platelet disorders. We investigated how these alterations cause distinct clinical phenotypes by combining structural, biochemical, and genomic approaches with gene complementation systems that examine GATA1 function in biologically relevant cellular contexts. Substitutions that disrupt FOG1 cofactor binding impair both gene activation and repression and are associated with pronounced clinical phenotypes. Moreover, clinical severity correlates with the degree of FOG1 disruption. Surprisingly, 2 mutations shown to impair DNA binding of GATA1 in vitro did not measurably affect in vivo target gene occupancy. Rather, one of these disrupted binding to the TAL1 complex, implicating it in diseases caused by GATA1 mutations. Diminished TAL1 complex recruitment mainly impairs transcriptional activation and is linked to relatively mild disease. Notably, different substitutions at the same amino acid can selectively inhibit TAL1 complex or FOG1 binding, producing distinct cellular and clinical phenotypes. The structure-function relationships elucidated here were not predicted by prior in vitro or computational studies. Thus, our findings uncover novel disease mechanisms underlying GATA1 mutations and highlight the power of gene complementation assays for elucidating the molecular basis of genetic diseases.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2012\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2012')\"\n      onkeypress=\"toggleGroup('group_2012')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2012</span>\n      <span class=\"bibbase_group_count\">\n        \n        (17)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012', 'http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111', event);\">\n    Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  McKenzie,  J., L.; Duyvestyn,  J., M.; Smith,  T.; Bendak,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Cursons,  R.; Cook,  G., M.;  and Arcus,  V., L.\n</span>\n\n  \n\n</span>\n\n  <i>RNA</i>, 18(6): 1267-1278. 6 2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111\"\n     onclick=\"log_download('mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012', 'http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1261/rna.031229.111\"\n     onclick=\"log_download('mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012', 'http://doi.org/10.1261/rna.031229.111', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry},\n type = {article},\n year = {2012},\n keywords = {Mycobacteria,PIN-domain,RNA interferase,RNase,Toxin-antitoxin,VapC},\n pages = {1267-1278},\n volume = {18},\n websites = {http://rnajournal.cshlp.org/cgi/doi/10.1261/rna.031229.111},\n month = {6},\n day = {1},\n id = {52a8f59e-942d-361e-8a56-5a3ea03f061e},\n created = {2020-12-17T05:29:53.840Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:53.840Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {McKenzie2012},\n source_type = {ARTICLE},\n notes = {cited By 31},\n private_publication = {false},\n abstract = {The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.},\n bibtype = {article},\n author = {McKenzie, J. L. and Duyvestyn, Johanna M. and Smith, Tony and Bendak, Katerina and MacKay, J. and Cursons, R. and Cook, Gregory M. and Arcus, Vickery L.},\n doi = {10.1261/rna.031229.111},\n journal = {RNA},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1002/pro.2124\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012', 'http://doi.wiley.com/10.1002/pro.2124', event);\">\n    Interdomain orientation of cardiac Troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Cordina,  N., M.; Liew,  C., K.; Gell,  D., A.; Fajer,  P., G.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Brown,  L., J.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 21(9): 1376-1387. 9 2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1002/pro.2124\"\n     onclick=\"log_download('cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012', 'http://doi.wiley.com/10.1002/pro.2124', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Interdomain orientation of cardiac Troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.2124\"\n     onclick=\"log_download('cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012', 'http://doi.org/10.1002/pro.2124', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Interdomain orientation of cardiac Troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state},\n type = {article},\n year = {2012},\n keywords = {Cardiac troponin C,Ensemble states,Paramagnetic relaxation enhancement,Site-directed spin labeling,Solution NMR},\n pages = {1376-1387},\n volume = {21},\n websites = {http://doi.wiley.com/10.1002/pro.2124},\n month = {9},\n id = {136eec2a-03b6-3cd9-bce5-f2eed2bdbc99},\n created = {2020-12-17T05:29:54.085Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:54.085Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Cordina2012},\n source_type = {ARTICLE},\n notes = {cited By 9},\n private_publication = {false},\n abstract = {Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.},\n bibtype = {article},\n author = {Cordina, Nicole M. and Liew, Chu Kong and Gell, David A. and Fajer, Piotr G. and Mackay, Joel P. and Brown, Louise J.},\n doi = {10.1002/pro.2124},\n journal = {Protein Science},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"morris-kwan-mackay-sunde-backboneandsidechain1h13cand15nchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://link.springer.com/10.1007/s12104-011-9330-5\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'morris-kwan-mackay-sunde-backboneandsidechain1h13cand15nchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012', 'http://link.springer.com/10.1007/s12104-011-9330-5', event);\">\n    Backbone and sidechain 1H, 13C and 15N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Morris,  V., K.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H.</a>; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Sunde,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 6(1): 83-86. 4 2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://link.springer.com/10.1007/s12104-011-9330-5\"\n     onclick=\"log_download('morris-kwan-mackay-sunde-backboneandsidechain1h13cand15nchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012', 'http://link.springer.com/10.1007/s12104-011-9330-5', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Backbone and sidechain 1H, 13C and 15N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-011-9330-5\"\n     onclick=\"log_download('morris-kwan-mackay-sunde-backboneandsidechain1h13cand15nchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012', 'http://doi.org/10.1007/s12104-011-9330-5', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/morris-kwan-mackay-sunde-backboneandsidechain1h13cand15nchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('morris-kwan-mackay-sunde-backboneandsidechain1h13cand15nchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Backbone and sidechain 1H, 13C and 15N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans},\n type = {article},\n year = {2012},\n keywords = {DewA,Functional amyloid,Hydrophobin,NMR assignment},\n pages = {83-86},\n volume = {6},\n websites = {http://link.springer.com/10.1007/s12104-011-9330-5},\n month = {4},\n day = {4},\n id = {852baf30-6704-30e4-8fdc-8cbe794bb64a},\n created = {2020-12-17T05:29:56.237Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:56.237Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Morris2012},\n source_type = {ARTICLE},\n notes = {cited By 4},\n private_publication = {false},\n abstract = {Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.},\n bibtype = {article},\n author = {Morris, Vanessa K. and Kwan, Ann H. and Mackay, Joel P. and Sunde, Margaret},\n doi = {10.1007/s12104-011-9330-5},\n journal = {Biomolecular NMR Assignments},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2lim2ldb1lidcomplex-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1002/pro.2153\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2lim2ldb1lidcomplex-2012', 'http://doi.wiley.com/10.1002/pro.2153', event);\">\n    Solution structure of a tethered Lmo2 LIM2 /Ldb1 LID complex.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dastmalchi,  S.; Wilkinson-White,  L.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H.</a>; Gamsjaeger,  R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Matthews,  J., M.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 21(11): 1768-1774. 11 2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1002/pro.2153\"\n     onclick=\"log_download('dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2lim2ldb1lidcomplex-2012', 'http://doi.wiley.com/10.1002/pro.2153', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Solution structure of a tethered Lmo2 LIM2 /Ldb1 LID complex [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.2153\"\n     onclick=\"log_download('dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2lim2ldb1lidcomplex-2012', 'http://doi.org/10.1002/pro.2153', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2lim2ldb1lidcomplex-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2lim2ldb1lidcomplex-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution structure of a tethered Lmo2 LIM2 /Ldb1 LID complex},\n type = {article},\n year = {2012},\n keywords = {Ldb1,Lmo2,Modular binding,NMR structure},\n pages = {1768-1774},\n volume = {21},\n websites = {http://doi.wiley.com/10.1002/pro.2153},\n month = {11},\n id = {574f81df-0620-3c4a-af49-474d48a6b829},\n created = {2020-12-17T05:29:56.294Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:56.294Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Dastmalchi2012},\n source_type = {ARTICLE},\n notes = {cited By 7},\n private_publication = {false},\n abstract = {LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.},\n bibtype = {article},\n author = {Dastmalchi, Siavoush and Wilkinson-White, Lorna and Kwan, Ann H. and Gamsjaeger, Roland and Mackay, Joel P. and Matthews, Jacqueline M.},\n doi = {10.1002/pro.2153},\n journal = {Protein Science},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"bhati-lee-gadd-jeffries-kwan-whitten-trewhella-mackay-etal-solutionstructureofthelimhomeodomaintranscriptionfactorcomplexlhx3ldb1andtheeffectsofapituitarymutationonkeylhx3interactions-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Solution structure of the LIM-homeodomain transcription factor complex Lhx3/Ldb1 and the effects of a pituitary mutation on key Lhx3 interactions.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Bhati,  M.; Lee,  C.; Gadd,  M.; Jeffries,  C.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Whitten,  A.; Trewhella,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>PLoS ONE</i>, 7(7).  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1371/journal.pone.0040719\"\n     onclick=\"log_download('bhati-lee-gadd-jeffries-kwan-whitten-trewhella-mackay-etal-solutionstructureofthelimhomeodomaintranscriptionfactorcomplexlhx3ldb1andtheeffectsofapituitarymutationonkeylhx3interactions-2012', 'http://doi.org/10.1371/journal.pone.0040719', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/bhati-lee-gadd-jeffries-kwan-whitten-trewhella-mackay-etal-solutionstructureofthelimhomeodomaintranscriptionfactorcomplexlhx3ldb1andtheeffectsofapituitarymutationonkeylhx3interactions-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('bhati-lee-gadd-jeffries-kwan-whitten-trewhella-mackay-etal-solutionstructureofthelimhomeodomaintranscriptionfactorcomplexlhx3ldb1andtheeffectsofapituitarymutationonkeylhx3interactions-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution structure of the LIM-homeodomain transcription factor complex Lhx3/Ldb1 and the effects of a pituitary mutation on key Lhx3 interactions},\n type = {article},\n year = {2012},\n volume = {7},\n id = {128d3304-f755-382e-92e5-6fc2f2229b4f},\n created = {2023-01-10T01:44:53.740Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:53.740Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Lhx3 is a LIM-homeodomain (LIM-HD) transcription factor that regulates neural cell subtype specification and pituitary development in vertebrates, and mutations in this protein cause combined pituitary hormone deficiency syndrome (CPHDS). The recently published structures of Lhx3 in complex with each of two key protein partners, Isl1 and Ldb1, provide an opportunity to understand the effect of mutations and posttranslational modifications on key protein-protein interactions. Here, we use small-angle X-ray scattering of an Ldb1-Lhx3 complex to confirm that in solution the protein is well represented by our previously determined NMR structure as an ensemble of conformers each comprising two well-defined halves (each made up of LIM domain from Lhx3 and the corresponding binding motif in Ldb1) with some flexibility between the two halves. NMR analysis of an Lhx3 mutant that causes CPHDS, Lhx3(Y114C), shows that the mutation does not alter the zinc-ligation properties of Lhx3, but appears to cause a structural rearrangement of the hydrophobic core of the LIM2 domain of Lhx3 that destabilises the domain and/or reduces the affinity of Lhx3 for both Ldb1 and Isl1. Thus the mutation would affect the formation of Lhx3-containing transcription factor complexes, particularly in the pituitary gland where these complexes are required for the production of multiple pituitary cell types and hormones. © 2012 Bhati et al.},\n bibtype = {article},\n author = {Bhati, M. and Lee, C. and Gadd, M.S. and Jeffries, C.M. and Kwan, A. and Whitten, A.E. and Trewhella, J. and Mackay, J.P. and Matthews, J.M.},\n doi = {10.1371/journal.pone.0040719},\n journal = {PLoS ONE},\n number = {7}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Lhx3 is a LIM-homeodomain (LIM-HD) transcription factor that regulates neural cell subtype specification and pituitary development in vertebrates, and mutations in this protein cause combined pituitary hormone deficiency syndrome (CPHDS). The recently published structures of Lhx3 in complex with each of two key protein partners, Isl1 and Ldb1, provide an opportunity to understand the effect of mutations and posttranslational modifications on key protein-protein interactions. Here, we use small-angle X-ray scattering of an Ldb1-Lhx3 complex to confirm that in solution the protein is well represented by our previously determined NMR structure as an ensemble of conformers each comprising two well-defined halves (each made up of LIM domain from Lhx3 and the corresponding binding motif in Ldb1) with some flexibility between the two halves. NMR analysis of an Lhx3 mutant that causes CPHDS, Lhx3(Y114C), shows that the mutation does not alter the zinc-ligation properties of Lhx3, but appears to cause a structural rearrangement of the hydrophobic core of the LIM2 domain of Lhx3 that destabilises the domain and/or reduces the affinity of Lhx3 for both Ldb1 and Isl1. Thus the mutation would affect the formation of Lhx3-containing transcription factor complexes, particularly in the pituitary gland where these complexes are required for the production of multiple pituitary cell types and hormones. © 2012 Bhati et al.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"bendak-loughlin-cheung-oconnell-crossley-mackay-arapidmethodforassessingthernabindingpotentialofaprotein-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A rapid method for assessing the RNA-binding potential of a protein.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Bendak,  K.; Loughlin,  F.; Cheung,  V.; O'Connell,  M.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Nucleic Acids Research</i>, 40(14).  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1093/nar/gks285\"\n     onclick=\"log_download('bendak-loughlin-cheung-oconnell-crossley-mackay-arapidmethodforassessingthernabindingpotentialofaprotein-2012', 'http://doi.org/10.1093/nar/gks285', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/bendak-loughlin-cheung-oconnell-crossley-mackay-arapidmethodforassessingthernabindingpotentialofaprotein-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('bendak-loughlin-cheung-oconnell-crossley-mackay-arapidmethodforassessingthernabindingpotentialofaprotein-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A rapid method for assessing the RNA-binding potential of a protein},\n type = {article},\n year = {2012},\n volume = {40},\n id = {45db7a3c-9c0b-3ecb-a0b6-8ca4c8c9c8cc},\n created = {2023-01-10T01:44:54.656Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:54.656Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures. © The Author(s) 2012.},\n bibtype = {article},\n author = {Bendak, K. and Loughlin, F.E. and Cheung, V. and O&#x27;Connell, M.R. and Crossley, M. and MacKay, J.P.},\n doi = {10.1093/nar/gks285},\n journal = {Nucleic Acids Research},\n number = {14}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures. © The Author(s) 2012.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"musselman-ramiez-sims-mansfield-oliver-denu-mackay-wade-etal-bivalentrecognitionofnucleosomesbythetandemphdfingersofthechd4atpaseisrequiredforchd4mediatedrepression-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Bivalent recognition of nucleosomes by the tandem PHD fingers of the CHD4 ATPase is required for CHD4-mediated repression.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Musselman,  C.; Ramiŕez,  J.; Sims,  J.; Mansfield,  R.; Oliver,  S.; Denu,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Wade,  P.; Hagman,  J.;  and Kutateladze,  T.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 109(3): 787-792.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.1113655109\"\n     onclick=\"log_download('musselman-ramiez-sims-mansfield-oliver-denu-mackay-wade-etal-bivalentrecognitionofnucleosomesbythetandemphdfingersofthechd4atpaseisrequiredforchd4mediatedrepression-2012', 'http://doi.org/10.1073/pnas.1113655109', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/musselman-ramiez-sims-mansfield-oliver-denu-mackay-wade-etal-bivalentrecognitionofnucleosomesbythetandemphdfingersofthechd4atpaseisrequiredforchd4mediatedrepression-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('musselman-ramiez-sims-mansfield-oliver-denu-mackay-wade-etal-bivalentrecognitionofnucleosomesbythetandemphdfingersofthechd4atpaseisrequiredforchd4mediatedrepression-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Bivalent recognition of nucleosomes by the tandem PHD fingers of the CHD4 ATPase is required for CHD4-mediated repression},\n type = {article},\n year = {2012},\n pages = {787-792},\n volume = {109},\n id = {fd6d1a67-ad29-356c-9c60-bd33f657e985},\n created = {2023-01-10T01:44:55.575Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:55.575Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {CHD4 is a catalytic subunit of the NuRD (nucleosome remodeling and deacetylase) complex essential in transcriptional regulation, chromatin assembly and DNA damage repair. CHD4 contains tandem plant homeodomain (PHD) fingers connected by a short linker, the biological function of which remains unclear. Here we explore the combinatorial action of the CHD4 PHD1/2 fingers and detail the molecular basis for their association with chromatin. We found that PHD1/2 targets nucleosomes in a multivalent manner, concomitantly engaging two histone H3 tails. This robust synergistic interaction displaces HP1γ from pericentric sites, inducing changes in chromatin structure and leading to the dispersion of the heterochromatic mark H3K9me3. We demonstrate that recognition of the histone H3 tails by the PHD fingers is required for repressive activity of the CHD4/NuRD complex. Together, our data elucidate the molecular mechanism of multivalent association of the PHD fingers with chromatin and reveal their critical role in the regulation of CHD4 functions.},\n bibtype = {article},\n author = {Musselman, C.A. and Ramiŕez, J. and Sims, J.K. and Mansfield, R.E. and Oliver, S.S. and Denu, J.M. and Mackay, J.P. and Wade, P.A. and Hagman, J. and Kutateladze, T.G.},\n doi = {10.1073/pnas.1113655109},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  CHD4 is a catalytic subunit of the NuRD (nucleosome remodeling and deacetylase) complex essential in transcriptional regulation, chromatin assembly and DNA damage repair. CHD4 contains tandem plant homeodomain (PHD) fingers connected by a short linker, the biological function of which remains unclear. Here we explore the combinatorial action of the CHD4 PHD1/2 fingers and detail the molecular basis for their association with chromatin. We found that PHD1/2 targets nucleosomes in a multivalent manner, concomitantly engaging two histone H3 tails. This robust synergistic interaction displaces HP1γ from pericentric sites, inducing changes in chromatin structure and leading to the dispersion of the heterochromatic mark H3K9me3. We demonstrate that recognition of the histone H3 tails by the PHD fingers is required for repressive activity of the CHD4/NuRD complex. Together, our data elucidate the molecular mechanism of multivalent association of the PHD fingers with chromatin and reveal their critical role in the regulation of CHD4 functions.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"burdach-oconnell-mackay-crossley-twotimingzincfingertranscriptionfactorsliaisingwithrna-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Two-timing zinc finger transcription factors liaising with RNA.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Burdach,  J.; O'Connell,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Trends in Biochemical Sciences</i>, 37(5): 199-205.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.tibs.2012.02.001\"\n     onclick=\"log_download('burdach-oconnell-mackay-crossley-twotimingzincfingertranscriptionfactorsliaisingwithrna-2012', 'http://doi.org/10.1016/j.tibs.2012.02.001', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/burdach-oconnell-mackay-crossley-twotimingzincfingertranscriptionfactorsliaisingwithrna-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('burdach-oconnell-mackay-crossley-twotimingzincfingertranscriptionfactorsliaisingwithrna-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Two-timing zinc finger transcription factors liaising with RNA},\n type = {article},\n year = {2012},\n pages = {199-205},\n volume = {37},\n id = {2554db33-7b4b-3127-99cf-3fde4951ed45},\n created = {2023-01-10T01:44:56.496Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:56.496Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Classical zinc fingers (ZFs) are one of the most common protein domains in higher eukaryotes and have been known for almost 30 years to act as sequence-specific DNA-binding domains. This knowledge has come, however, from the study of a small number of archetypal proteins, and a larger picture is beginning to emerge that ZF functions are far more diverse than originally suspected. Here, we review the evidence that a subset of ZF proteins live double lives, binding to both DNA and RNA targets and frequenting both the cytoplasm and the nucleus. This duality can create an important additional level of gene regulation that serves to connect transcriptional and post-transcriptional control. © 2012 Elsevier Ltd.},\n bibtype = {article},\n author = {Burdach, J. and O&#x27;Connell, M.R. and Mackay, J.P. and Crossley, M.},\n doi = {10.1016/j.tibs.2012.02.001},\n journal = {Trends in Biochemical Sciences},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Classical zinc fingers (ZFs) are one of the most common protein domains in higher eukaryotes and have been known for almost 30 years to act as sequence-specific DNA-binding domains. This knowledge has come, however, from the study of a small number of archetypal proteins, and a larger picture is beginning to emerge that ZF functions are far more diverse than originally suspected. Here, we review the evidence that a subset of ZF proteins live double lives, binding to both DNA and RNA targets and frequenting both the cytoplasm and the nucleus. This duality can create an important additional level of gene regulation that serves to connect transcriptional and post-transcriptional control. © 2012 Elsevier Ltd.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2inflim2infldb1inflidinfcomplex-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Solution structure of a tethered Lmo2<inf>LIM2</inf>/Ldb1<inf>LID</inf> complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dastmalchi,  S.; Wilkinson-White,  L.; Kwan,  A.; Gamsjaeger,  R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 21(11): 1768-1774.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.2153\"\n     onclick=\"log_download('dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2inflim2infldb1inflidinfcomplex-2012', 'http://doi.org/10.1002/pro.2153', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2inflim2infldb1inflidinfcomplex-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dastmalchi-wilkinsonwhite-kwan-gamsjaeger-mackay-matthews-solutionstructureofatetheredlmo2inflim2infldb1inflidinfcomplex-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution structure of a tethered Lmo2&lt;inf&gt;LIM2&lt;/inf&gt;/Ldb1&lt;inf&gt;LID&lt;/inf&gt; complex},\n type = {article},\n year = {2012},\n pages = {1768-1774},\n volume = {21},\n id = {c8d8b2ce-e804-3b1e-bc6d-dea5554342b7},\n created = {2023-01-10T01:44:57.390Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:57.390Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.},\n bibtype = {article},\n author = {Dastmalchi, S. and Wilkinson-White, L. and Kwan, A.H. and Gamsjaeger, R. and Mackay, J.P. and Matthews, J.M.},\n doi = {10.1002/pro.2153},\n journal = {Protein Science},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  LIM-only protein 2, Lmo2, is a regulatory protein that is essential for hematopoietic development and inappropriate overexpression of Lmo2 in T-cells contributes to T-cell leukemia. It exerts its functions by mediating protein-protein interactions and nucleating multicomponent transcriptional complexes. Lmo2 interacts with LIM domain binding protein 1 (Ldb1) through the tandem LIM domains of Lmo2 and the LIM interaction domain (LID) of Ldb1. Here, we present the solution structure of the LIM2 domain of Lmo2 bound to Ldb1 LID. The ordered regions of Ldb1 in this complex correspond well with binding hotspots previously defined by mutagenic studies. Comparisons of this Lmo2LIM2-Ldb1LID structure with previously determined structures of the Lmo2/Ldb1LID complexes lead to the conclusion that modular binding of tandem LIM domains in Lmo2 to tandem linear motifs in Ldb1 is accompanied by several disorder-to-order transitions and/or conformational changes in both proteins. © 2012 The Protein Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"oconnell-vandevenne-nguyen-matthews-gamsjaeger-segal-mackay-modularassemblyofranbp2typezincfingerdomainstotargetsinglestrandedrna-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Modular assembly of RanBP2-Type zinc finger domains to target single-stranded RNA.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  O'Connell,  M.; Vandevenne,  M.; Nguyen,  C.; Matthews,  J.; Gamsjaeger,  R.; Segal,  D.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Angewandte Chemie - International Edition</i>, 51(22): 5371-5375.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/anie.201200866\"\n     onclick=\"log_download('oconnell-vandevenne-nguyen-matthews-gamsjaeger-segal-mackay-modularassemblyofranbp2typezincfingerdomainstotargetsinglestrandedrna-2012', 'http://doi.org/10.1002/anie.201200866', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/oconnell-vandevenne-nguyen-matthews-gamsjaeger-segal-mackay-modularassemblyofranbp2typezincfingerdomainstotargetsinglestrandedrna-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('oconnell-vandevenne-nguyen-matthews-gamsjaeger-segal-mackay-modularassemblyofranbp2typezincfingerdomainstotargetsinglestrandedrna-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Modular assembly of RanBP2-Type zinc finger domains to target single-stranded RNA},\n type = {article},\n year = {2012},\n pages = {5371-5375},\n volume = {51},\n id = {524d43b0-eac3-303b-9076-d08253f97d3b},\n created = {2023-01-10T01:44:58.301Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:58.301Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Fingering RNA: To study the function of the variety of RNA species that have been discovered recently, sequence-specific RNA-binding molecules with tunable specificity are required. Here, the use of zinc fingers (ZFs) from the splicing factors ZRANB2 and RBM5 as potential modules for the assembly of sequence-specific RNA-binding molecules is demonstrated. Copyright © 2012 WILEY-VCH Verlag GmbH &amp; Co. KGaA, Weinheim.},\n bibtype = {article},\n author = {O&#x27;Connell, M.R. and Vandevenne, M. and Nguyen, C.D. and Matthews, J.M. and Gamsjaeger, R. and Segal, D.J. and MacKay, J.P.},\n doi = {10.1002/anie.201200866},\n journal = {Angewandte Chemie - International Edition},\n number = {22}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Fingering RNA: To study the function of the variety of RNA species that have been discovered recently, sequence-specific RNA-binding molecules with tunable specificity are required. Here, the use of zinc fingers (ZFs) from the splicing factors ZRANB2 and RBM5 as potential modules for the assembly of sequence-specific RNA-binding molecules is demonstrated. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"liew-kwan-stokes-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo4lim1ctipcomplex-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular LMO4-LIM1/CtIP complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Liew,  C.; Kwan,  A.; Stokes,  P.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 6(1): 31-34.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-011-9319-0\"\n     onclick=\"log_download('liew-kwan-stokes-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo4lim1ctipcomplex-2012', 'http://doi.org/10.1007/s12104-011-9319-0', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/liew-kwan-stokes-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo4lim1ctipcomplex-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('liew-kwan-stokes-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo4lim1ctipcomplex-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {&lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;15&lt;/sup&gt;N and &lt;sup&gt;13&lt;/sup&gt;C assignments of an intramolecular LMO4-LIM1/CtIP complex},\n type = {article},\n year = {2012},\n pages = {31-34},\n volume = {6},\n id = {f121ae58-5c4e-3fbb-987c-35bd8fe224e1},\n created = {2023-01-10T01:44:59.248Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:44:59.248Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {LMO4 is a broadly expressed LIM-only protein that is involved in neural tube development and implicated in breast cancer. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the N-terminal LIM domain from LMO4 tethered to residues 641-685 of C-terminal binding protein interacting protein (CtIP/RBBP8). © 2011 Springer Science+Business Media B.V.},\n bibtype = {article},\n author = {Liew, C.W. and Kwan, A.H. and Stokes, P.H. and MacKay, J.P. and Matthews, J.M.},\n doi = {10.1007/s12104-011-9319-0},\n journal = {Biomolecular NMR Assignments},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  LMO4 is a broadly expressed LIM-only protein that is involved in neural tube development and implicated in breast cancer. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the N-terminal LIM domain from LMO4 tethered to residues 641-685 of C-terminal binding protein interacting protein (CtIP/RBBP8). © 2011 Springer Science+Business Media B.V.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"hsieh-wilkinson-oconnell-mackay-matthews-payne-synthesisofthebacteriocinglycopeptidesublancin168andsglycosylatedvariants-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Synthesis of the bacteriocin glycopeptide sublancin 168 and S-glycosylated variants.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Hsieh,  Y.; Wilkinson,  B.; O'Connell,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Matthews,  J.;  and Payne,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Organic Letters</i>, 14(7): 1910-1913.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/ol300557g\"\n     onclick=\"log_download('hsieh-wilkinson-oconnell-mackay-matthews-payne-synthesisofthebacteriocinglycopeptidesublancin168andsglycosylatedvariants-2012', 'http://doi.org/10.1021/ol300557g', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/hsieh-wilkinson-oconnell-mackay-matthews-payne-synthesisofthebacteriocinglycopeptidesublancin168andsglycosylatedvariants-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('hsieh-wilkinson-oconnell-mackay-matthews-payne-synthesisofthebacteriocinglycopeptidesublancin168andsglycosylatedvariants-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Synthesis of the bacteriocin glycopeptide sublancin 168 and S-glycosylated variants},\n type = {article},\n year = {2012},\n pages = {1910-1913},\n volume = {14},\n id = {0782aeb9-904a-31dc-bf32-5d68b28b7eb4},\n created = {2023-01-10T01:45:00.240Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:00.240Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The synthesis of sublancin 168, a unique S-glucosylated bacteriocin antibiotic, is described. The natural product and two S-glycosylated variants were successfully prepared via native chemical ligation followed by folding. The synthetic glycopeptides were shown to possess primarily an α-helical secondary structure by CD and NMR studies. © 2012 American Chemical Society.},\n bibtype = {article},\n author = {Hsieh, Y.S.Y. and Wilkinson, B.L. and O&#x27;Connell, M.R. and MacKay, J.P. and Matthews, J.M. and Payne, R.J.},\n doi = {10.1021/ol300557g},\n journal = {Organic Letters},\n number = {7}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The synthesis of sublancin 168, a unique S-glucosylated bacteriocin antibiotic, is described. The natural product and two S-glycosylated variants were successfully prepared via native chemical ligation followed by folding. The synthetic glycopeptides were shown to possess primarily an α-helical secondary structure by CD and NMR studies. © 2012 American Chemical Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"morris-kwan-mackay-sunde-backboneandsidechainsup1suphsup13supcandsup15supnchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Backbone and sidechain <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Morris,  V.; Kwan,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Sunde,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 6(1): 83-86.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-011-9330-5\"\n     onclick=\"log_download('morris-kwan-mackay-sunde-backboneandsidechainsup1suphsup13supcandsup15supnchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012', 'http://doi.org/10.1007/s12104-011-9330-5', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/morris-kwan-mackay-sunde-backboneandsidechainsup1suphsup13supcandsup15supnchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('morris-kwan-mackay-sunde-backboneandsidechainsup1suphsup13supcandsup15supnchemicalshiftassignmentsofthehydrophobindewafromaspergillusnidulans-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Backbone and sidechain &lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;13&lt;/sup&gt;C and &lt;sup&gt;15&lt;/sup&gt;N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans},\n type = {article},\n year = {2012},\n pages = {83-86},\n volume = {6},\n id = {ff9a844c-9cab-30ff-b896-a298653e06c7},\n created = {2023-01-10T01:45:01.153Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:01.153Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.},\n bibtype = {article},\n author = {Morris, V.K. and Kwan, A.H. and MacKay, J.P. and Sunde, M.},\n doi = {10.1007/s12104-011-9330-5},\n journal = {Biomolecular NMR Assignments},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Hydrophobins are proteins secreted by filamentous fungi that are able to self-assemble into monolayers at hydrophobic:hydrophilic interfaces. The layers are amphipathic and can reverse the wettability of surfaces. Hydrophobins have several roles in fungal development, including the formation of coatings on fungal structures to render them hydrophobic. Here we report the backbone and sidechain assignments for the class I hydrophobin DewA from the fungus Aspergillus nidulans. © 2011 Springer Science+Business Media B.V.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mckenzie,  J.; Duyvestyn,  J.; Smith,  T.; Bendak,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Cursons,  R.; Cook,  G.;  and Arcus,  V.\n</span>\n\n  \n\n</span>\n\n  <i>RNA</i>, 18(6): 1267-1278.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1261/rna.031229.111\"\n     onclick=\"log_download('mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012', 'http://doi.org/10.1261/rna.031229.111', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mckenzie-duyvestyn-smith-bendak-mackay-cursons-cook-arcus-determinationofribonucleasesequencespecificityusingpentaprobesandmassspectrometry-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry},\n type = {article},\n year = {2012},\n pages = {1267-1278},\n volume = {18},\n id = {620041f4-f32a-3709-9ae9-50c212e6b3e8},\n created = {2023-01-10T01:45:02.072Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:02.072Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.},\n bibtype = {article},\n author = {Mckenzie, J.L. and Duyvestyn, J.M. and Smith, T. and Bendak, K. and Mackay, J. and Cursons, R.A.Y. and Cook, G.M. and Arcus, V.L.},\n doi = {10.1261/rna.031229.111},\n journal = {RNA},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press. Copyright © 2012 RNA Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Interdomain orientation of cardiac troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Cordina,  N.; Liew,  C.; Gell,  D.; Fajer,  P.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Brown,  L.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 21(9): 1376-1387.  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pro.2124\"\n     onclick=\"log_download('cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012', 'http://doi.org/10.1002/pro.2124', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('cordina-liew-gell-fajer-mackay-brown-interdomainorientationofcardiactroponinccharacterizedbyparamagneticrelaxationenhancementnmrrevealsacompactstate-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Interdomain orientation of cardiac troponin C characterized by paramagnetic relaxation enhancement NMR reveals a compact state},\n type = {article},\n year = {2012},\n pages = {1376-1387},\n volume = {21},\n id = {4e767ca7-1e6a-3aae-aa86-c051dd322f85},\n created = {2023-01-10T01:45:03.028Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:03.028Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.},\n bibtype = {article},\n author = {Cordina, N.M. and Liew, C.K. and Gell, D.A. and Fajer, P.G. and Mackay, J.P. and Brown, L.J.},\n doi = {10.1002/pro.2124},\n journal = {Protein Science},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Cardiac troponin C (cTnC) is the calcium binding subunit of the troponin complex that triggers the thin filament response to calcium influx into the sarcomere. cTnC consists of two globular EF-hand domains (termed the N- and C-domains) connected by a flexible linker. While the conformation of each domain of cTnC has been thoroughly characterized through NMR studies involving either the isolated N-domain (N-cTnC) or C-domain (C-cTnC), little attention has been paid to the range of interdomain orientations possible in full-length cTnC that arises as a consequence of the flexibility of the domain linker. Flexibility in the domain linker of cTnC is essential for effective regulatory function of troponin. We have therefore utilized paramagnetic relaxation enhancement (PRE) NMR to assess the interdomain orientation of cTnC. Ensemble fitting of our interdomain PRE measurements reveals that isolated cTnC has considerable interdomain flexibility and preferentially adopts a bent conformation in solution, with a defined range of relative domain orientations. Published by Wiley-Blackwell. © 2012 The Protein Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"macindoe-kwan-ren-morris-yang-mackay-sunde-selfassemblyoffunctionalamphipathicamyloidmonolayersbythefungalhydrophobineas-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Self-assembly of functional, amphipathic amyloid monolayers by the fungal hydrophobin EAS.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Macindoe,  I.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Ren,  Q.; Morris,  V.; Yang,  W.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Sunde,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 109(14).  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.1114052109\"\n     onclick=\"log_download('macindoe-kwan-ren-morris-yang-mackay-sunde-selfassemblyoffunctionalamphipathicamyloidmonolayersbythefungalhydrophobineas-2012', 'http://doi.org/10.1073/pnas.1114052109', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/macindoe-kwan-ren-morris-yang-mackay-sunde-selfassemblyoffunctionalamphipathicamyloidmonolayersbythefungalhydrophobineas-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('macindoe-kwan-ren-morris-yang-mackay-sunde-selfassemblyoffunctionalamphipathicamyloidmonolayersbythefungalhydrophobineas-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Self-assembly of functional, amphipathic amyloid monolayers by the fungal hydrophobin EAS},\n type = {article},\n year = {2012},\n volume = {109},\n id = {b585d9ee-1807-3f19-9891-d5c72141f251},\n created = {2023-01-10T01:45:04.060Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:04.060Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The hydrophobin EAS from the fungus Neurospora crassa forms functional amyloid fibrils called rodlets that facilitate spore formation and dispersal. Self-assembly of EAS into fibrillar rodlets occurs spontaneously at hydrophobic:hydrophilic interfaces and the rodlets further associate laterally to form amphipathic monolayers. We have used site-directed mutagenesis and peptide experiments to identify the region of EAS that drives intermolecular association and formation of the cross-β rodlet structure. Transplanting this region into a nonamyloidogenic hydrophobin enables it to form rodlets. We have also determined the structure and dynamics of an EAS variant with reduced rodlet-forming ability. Taken together, these data allow us to pinpoint the conformational changes that take place when hydrophobins self-assemble at an interface and to propose a model for the amphipathic EAS rodlet structure.},\n bibtype = {article},\n author = {Macindoe, I. and Kwan, A.H. and Ren, Q. and Morris, V.K. and Yang, W. and Mackay, J.P. and Sunde, M.},\n doi = {10.1073/pnas.1114052109},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {14}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The hydrophobin EAS from the fungus Neurospora crassa forms functional amyloid fibrils called rodlets that facilitate spore formation and dispersal. Self-assembly of EAS into fibrillar rodlets occurs spontaneously at hydrophobic:hydrophilic interfaces and the rodlets further associate laterally to form amphipathic monolayers. We have used site-directed mutagenesis and peptide experiments to identify the region of EAS that drives intermolecular association and formation of the cross-β rodlet structure. Transplanting this region into a nonamyloidogenic hydrophobin enables it to form rodlets. We have also determined the structure and dynamics of an EAS variant with reduced rodlet-forming ability. Taken together, these data allow us to pinpoint the conformational changes that take place when hydrophobins self-assemble at an interface and to propose a model for the amphipathic EAS rodlet structure.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gell-grant-mackay-thedetectionandquantitationofproteinoligomerization-2012\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The detection and quantitation of protein oligomerization.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gell,  D.; Grant,  R.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  Volume 747  2012.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/978-1-4614-3229-6_2\"\n     onclick=\"log_download('gell-grant-mackay-thedetectionandquantitationofproteinoligomerization-2012', 'http://doi.org/10.1007/978-1-4614-3229-6_2', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gell-grant-mackay-thedetectionandquantitationofproteinoligomerization-2012\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gell-grant-mackay-thedetectionandquantitationofproteinoligomerization-2012')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@book{\n title = {The detection and quantitation of protein oligomerization},\n type = {book},\n year = {2012},\n source = {Advances in Experimental Medicine and Biology},\n pages = {19-41},\n volume = {747},\n id = {7e3e3789-bc90-34ed-805a-f49344268f6e},\n created = {2023-01-10T01:45:04.982Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:04.982Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {There are many different techniques available to biologists and biochemists that can be used to detect and characterize the self-association of proteins. Each technique has strengths and weaknesses and it is often useful to combine several approaches to maximize the former and minimize the latter. Here we review a range of methodologies that identify protein self-association and/or allow the stoichiometry and affinity of the interaction to be determined, placing an emphasis on what type of information can be obtained and outlining the advantages and disadvantages involved. In general, in vitro biophysical techniques, such as size exclusion chromatography, analytical ultracentrifugation, scattering techniques, NMR spectroscopy, isothermal titration calorimetry, fluorescence anisotropy and mass spectrometry, provide information on stoichiometry and/or binding affinities. Other approaches such as cross-linking, fluorescence methods (e.g., fluorescence correlation spectroscopy, FCS; Förster resonance energy transfer, FRET; fluorescence recovery after photobleaching, FRAP; and proximity imaging, PRIM) and complementation approaches (e.g., yeast two hybrid assays and bimolecular fluorescence complementation, BiFC) can be used to detect protein self-association in a cellular context. © 2012 Springer Science+Business Media, LLC.},\n bibtype = {book},\n author = {Gell, D.A. and Grant, R.P. and MacKay, J.P.},\n doi = {10.1007/978-1-4614-3229-6_2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  There are many different techniques available to biologists and biochemists that can be used to detect and characterize the self-association of proteins. Each technique has strengths and weaknesses and it is often useful to combine several approaches to maximize the former and minimize the latter. Here we review a range of methodologies that identify protein self-association and/or allow the stoichiometry and affinity of the interaction to be determined, placing an emphasis on what type of information can be obtained and outlining the advantages and disadvantages involved. In general, in vitro biophysical techniques, such as size exclusion chromatography, analytical ultracentrifugation, scattering techniques, NMR spectroscopy, isothermal titration calorimetry, fluorescence anisotropy and mass spectrometry, provide information on stoichiometry and/or binding affinities. Other approaches such as cross-linking, fluorescence methods (e.g., fluorescence correlation spectroscopy, FCS; Förster resonance energy transfer, FRET; fluorescence recovery after photobleaching, FRAP; and proximity imaging, PRIM) and complementation approaches (e.g., yeast two hybrid assays and bimolecular fluorescence complementation, BiFC) can be used to detect protein self-association in a cellular context. © 2012 Springer Science+Business Media, LLC.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2011\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2011')\"\n      onkeypress=\"toggleGroup('group_2011')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2011</span>\n      <span class=\"bibbase_group_count\">\n        \n        (16)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011', 'http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108', event);\">\n    Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wilkinson-White,  L.; Gamsjaeger,  R.; Dastmalchi,  S.; Wienert,  B.; Stokes,  P., H.; Crossley,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Matthews,  J., M.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences</i>, 108(35): 14443-14448. 8 2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108\"\n     onclick=\"log_download('wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011', 'http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1 [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.1105898108\"\n     onclick=\"log_download('wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011', 'http://doi.org/10.1073/pnas.1105898108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1},\n type = {article},\n year = {2011},\n keywords = {Haematopoiesis,Protein-DNA interactions,Protein-protein interactions,Transcription factor complex},\n pages = {14443-14448},\n volume = {108},\n websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.1105898108},\n month = {8},\n day = {30},\n id = {a786b1a3-bd30-3c66-8aff-7ffb5cbbb0da},\n created = {2020-12-17T05:29:55.512Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:55.512Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Wilkinson-White2011},\n source_type = {ARTICLE},\n notes = {cited By 24},\n private_publication = {false},\n abstract = {The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (&lt;50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.},\n bibtype = {article},\n author = {Wilkinson-White, Lorna and Gamsjaeger, Roland and Dastmalchi, Siavoush and Wienert, Beeke and Stokes, Philippa H. and Crossley, Merlin and Mackay, Joel P. and Matthews, Jacqueline M.},\n doi = {10.1073/pnas.1105898108},\n journal = {Proceedings of the National Academy of Sciences},\n number = {35}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (<50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011', 'http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108', event);\">\n    Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Lamonica,  J., M.; Deng,  W.; Kadauke,  S.; Campbell,  A., E.; Gamsjaeger,  R.; Wang,  H.; Cheng,  Y.; Billin,  A., N.; Hardison,  R., C.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Blobel,  G., A.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences</i>, 108(22): E159-E168. 5 2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108\"\n     onclick=\"log_download('lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011', 'http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.1102140108\"\n     onclick=\"log_download('lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011', 'http://doi.org/10.1073/pnas.1102140108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes},\n type = {article},\n year = {2011},\n keywords = {Gene regulation,Hematopoiesis,Posttranslational modifications},\n pages = {E159-E168},\n volume = {108},\n websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.1102140108},\n month = {5},\n day = {31},\n id = {f5506c6c-a1c6-37df-8e6d-aee9893f4e46},\n created = {2020-12-17T05:29:57.064Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:57.064Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Lamonica2011},\n source_type = {ARTICLE},\n notes = {cited By 98},\n private_publication = {false},\n abstract = {Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 &quot;reads&quot; acetyl marks on nuclear factors to promote their stable association with chromatin.},\n bibtype = {article},\n author = {Lamonica, Janine M. and Deng, Wulan and Kadauke, Stephan and Campbell, Amy E. and Gamsjaeger, Roland and Wang, Hongxin and Cheng, Yong and Billin, Andrew N. and Hardison, Ross C. and Mackay, Joel P. and Blobel, Gerd A.},\n doi = {10.1073/pnas.1102140108},\n journal = {Proceedings of the National Academy of Sciences},\n number = {22}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 \"reads\" acetyl marks on nuclear factors to promote their stable association with chromatin.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"nunez-clifton-funnell-artuz-hallal-quinlan-font-vandevenne-etal-themultizincfingerproteinznf217contactsdnathroughatwofingerdomain-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The multi-zinc finger protein ZNF217 contacts DNA through a two-finger domain.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Nunez,  N.; Clifton,  M.; Funnell,  A.; Artuz,  C.; Hallal,  S.; Quinlan,  K.; Font,  J.; Vandevenne,  M.; Setiyaputra,  S.; Pearson,  R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 286(44): 38190-38201.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M111.301234\"\n     onclick=\"log_download('nunez-clifton-funnell-artuz-hallal-quinlan-font-vandevenne-etal-themultizincfingerproteinznf217contactsdnathroughatwofingerdomain-2011', 'http://doi.org/10.1074/jbc.M111.301234', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/nunez-clifton-funnell-artuz-hallal-quinlan-font-vandevenne-etal-themultizincfingerproteinznf217contactsdnathroughatwofingerdomain-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('nunez-clifton-funnell-artuz-hallal-quinlan-font-vandevenne-etal-themultizincfingerproteinznf217contactsdnathroughatwofingerdomain-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The multi-zinc finger protein ZNF217 contacts DNA through a two-finger domain},\n type = {article},\n year = {2011},\n pages = {38190-38201},\n volume = {286},\n id = {e1fdc7f2-dcf3-344a-b417-57036c8d3fcc},\n created = {2023-01-10T01:45:05.935Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:05.935Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Classical C2H2 zinc finger proteins are among the most abundant transcription factors found in eukaryotes, and the mechanisms through which they recognize their target genes have been extensively investigated. In general, a tandem array of three fingers separated by characteristic TGERP links is required for sequence-specific DNA recognition. Nevertheless, a significant number of zinc finger proteins do not contain a hallmark three-finger array of this type, raising the question of whether and how they contact DNA. We have examined the multi-finger protein ZNF217, which contains eight classical zinc fingers. ZNF217 is implicated as an oncogene and in repressing the E-cadherin gene. We show that two of its zinc fingers, 6 and 7, can mediate contacts with DNA. We examine its putative recognition site in the E-cadherin promoter and demonstrate that this is a suboptimal site. NMR analysis and mutagenesis is used to define the DNA binding surface of ZNF217, and we examine the specificity of the DNA binding activity using fluorescence anisotropy titrations. Finally, sequence analysis reveals that a variety of multi-finger proteins also contain two-finger units, and our data support the idea that these may constitute a distinct subclass of DNA recognition motif. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A.},\n bibtype = {article},\n author = {Nunez, N. and Clifton, M.M.K. and Funnell, A.P.W. and Artuz, C. and Hallal, S. and Quinlan, K.G.R. and Font, J. and Vandevenne, M. and Setiyaputra, S. and Pearson, R.C.M. and Mackay, J.P. and Crossley, M.},\n doi = {10.1074/jbc.M111.301234},\n journal = {Journal of Biological Chemistry},\n number = {44}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Classical C2H2 zinc finger proteins are among the most abundant transcription factors found in eukaryotes, and the mechanisms through which they recognize their target genes have been extensively investigated. In general, a tandem array of three fingers separated by characteristic TGERP links is required for sequence-specific DNA recognition. Nevertheless, a significant number of zinc finger proteins do not contain a hallmark three-finger array of this type, raising the question of whether and how they contact DNA. We have examined the multi-finger protein ZNF217, which contains eight classical zinc fingers. ZNF217 is implicated as an oncogene and in repressing the E-cadherin gene. We show that two of its zinc fingers, 6 and 7, can mediate contacts with DNA. We examine its putative recognition site in the E-cadherin promoter and demonstrate that this is a suboptimal site. NMR analysis and mutagenesis is used to define the DNA binding surface of ZNF217, and we examine the specificity of the DNA binding activity using fluorescence anisotropy titrations. Finally, sequence analysis reveals that a variety of multi-finger proteins also contain two-finger units, and our data support the idea that these may constitute a distinct subclass of DNA recognition motif. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Lamonica,  J.; Deng,  W.; Kadauke,  S.; Campbell,  A.; Gamsjaeger,  R.; Wang,  H.; Cheng,  Y.; Billin,  A.; Hardison,  R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Blobel,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 108(22).  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.1102140108\"\n     onclick=\"log_download('lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011', 'http://doi.org/10.1073/pnas.1102140108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('lamonica-deng-kadauke-campbell-gamsjaeger-wang-cheng-billin-etal-bromodomainproteinbrd3associateswithacetylatedgata1topromoteitschromatinoccupancyaterythroidtargetgenes-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes},\n type = {article},\n year = {2011},\n volume = {108},\n id = {cd0868d1-a20c-3300-8452-75a3d9552903},\n created = {2023-01-10T01:45:06.903Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:06.903Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 &quot;reads&quot; acetyl marks on nuclear factors to promote their stable association with chromatin.},\n bibtype = {article},\n author = {Lamonica, J.M. and Deng, W. and Kadauke, S. and Campbell, A.E. and Gamsjaeger, R. and Wang, H. and Cheng, Y. and Billin, A.N. and Hardison, R.C. and Mackay, J.P. and Mackay, J.P. and Blobel, G.A.},\n doi = {10.1073/pnas.1102140108},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {22}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Acetylation of histones triggers association with bromodomaincontaining proteins that regulate diverse chromatin-related processes. Although acetylation of transcription factors has been appreciated for some time, the mechanistic consequences are less well understood. The hematopoietic transcription factor GATA1 is acetylated at conserved lysines that are required for its stable association with chromatin. We show that the BET family protein Brd3 binds via its first bromodomain (BD1) to GATA1 in an acetylation-dependent manner in vitro and in vivo. Mutation of a single residue in BD1 that is involved in acetyl-lysine binding abrogated recruitment of Brd3 by GATA1, demonstrating that acetylation of GATA1 is essential for Brd3 association with chromatin. Notably, Brd3 is recruited by GATA1 to both active and repressed target genes in a fashion seemingly independent of histone acetylation. Anti-Brd3 ChIP followed by massively parallel sequencing in GATA1-deficient erythroid precursor cells and those that are GATA1 replete qrevealed that GATA1 is a major determinant of Brd3 recruitment to genomic targets within chromatin. A pharmacologic compound that occupies the acetyl-lysine binding pockets of Brd3 bromodomains disrupts the Brd3-GATA1 interaction, diminishes the chromatin occupancy of both proteins, and inhibits erythroid maturation. Together these findings provide a mechanism for GATA1 acetylation and suggest that Brd3 \"reads\" acetyl marks on nuclear factors to promote their stable association with chromatin.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kumar-jacques-pishchany-caradocdavies-spirig-malmirchegini-langley-dickson-etal-structuralbasisforhemoglobincapturebystaphylococcusaureuscellsurfaceproteinisdh-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural basis for hemoglobin capture by Staphylococcus aureus cell-surface protein, IsdH.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kumar,  K.; Jacques,  D.; Pishchany,  G.; Caradoc-Davies,  T.; Spirig,  T.; Malmirchegini,  G.; Langley,  D.; Dickson,  C.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Clubb,  R.; Guss,  J.;  and Gell,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 286(44): 38439-38447.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M111.287300\"\n     onclick=\"log_download('kumar-jacques-pishchany-caradocdavies-spirig-malmirchegini-langley-dickson-etal-structuralbasisforhemoglobincapturebystaphylococcusaureuscellsurfaceproteinisdh-2011', 'http://doi.org/10.1074/jbc.M111.287300', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kumar-jacques-pishchany-caradocdavies-spirig-malmirchegini-langley-dickson-etal-structuralbasisforhemoglobincapturebystaphylococcusaureuscellsurfaceproteinisdh-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kumar-jacques-pishchany-caradocdavies-spirig-malmirchegini-langley-dickson-etal-structuralbasisforhemoglobincapturebystaphylococcusaureuscellsurfaceproteinisdh-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural basis for hemoglobin capture by Staphylococcus aureus cell-surface protein, IsdH},\n type = {article},\n year = {2011},\n pages = {38439-38447},\n volume = {286},\n id = {206f34b7-fe09-3a3c-a885-250484075b13},\n created = {2023-01-10T01:45:07.824Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:07.824Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Pathogens must steal iron from their hosts to establish infection. In mammals, hemoglobin (Hb) represents the largest reservoir of iron, and pathogens express Hb-binding proteins to access this source. Here, we show how one of the commonest and most significant human pathogens, Staphylococcus aureus, captures Hb as the first step of an iron-scavenging pathway. The x-ray crystal structure of Hb bound to a domain from the Isd (iron-regulated surface determinant) protein, IsdH, is the first structure of a Hb capture complex to be determined. Surface mutations in Hb that reduce binding to the Hb-receptor limit the capacity of S. aureus to utilize Hb as an iron source, suggesting that Hb sequence is a factor in host susceptibility to infection. The demonstration that pathogens make highly specific recognition complexes with Hb raises the possibility of developing inhibitors of Hb binding as antibacterial agents. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Kumar, K.K. and Jacques, D.A. and Pishchany, G. and Caradoc-Davies, T. and Spirig, T. and Malmirchegini, G.R. and Langley, D.B. and Dickson, C.F. and Mackay, J.P. and Clubb, R.T. and Guss, J.M. and Gell, D.A.},\n doi = {10.1074/jbc.M111.287300},\n journal = {Journal of Biological Chemistry},\n number = {44}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Pathogens must steal iron from their hosts to establish infection. In mammals, hemoglobin (Hb) represents the largest reservoir of iron, and pathogens express Hb-binding proteins to access this source. Here, we show how one of the commonest and most significant human pathogens, Staphylococcus aureus, captures Hb as the first step of an iron-scavenging pathway. The x-ray crystal structure of Hb bound to a domain from the Isd (iron-regulated surface determinant) protein, IsdH, is the first structure of a Hb capture complex to be determined. Surface mutations in Hb that reduce binding to the Hb-receptor limit the capacity of S. aureus to utilize Hb as an iron source, suggesting that Hb sequence is a factor in host susceptibility to infection. The demonstration that pathogens make highly specific recognition complexes with Hb raises the possibility of developing inhibitors of Hb binding as antibacterial agents. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wissmueller-font-liew-cram-schroeder-turner-crossley-mackay-etal-proteinproteininteractionsanalysisofafalsepositivegstpulldownresult-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Protein-protein interactions: Analysis of a false positive GST pulldown result.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wissmueller,  S.; Font,  J.; Liew,  C.; Cram,  E.; Schroeder,  T.; Turner,  J.; Crossley,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Proteins: Structure, Function and Bioinformatics</i>, 79(8): 2365-2371.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/prot.23068\"\n     onclick=\"log_download('wissmueller-font-liew-cram-schroeder-turner-crossley-mackay-etal-proteinproteininteractionsanalysisofafalsepositivegstpulldownresult-2011', 'http://doi.org/10.1002/prot.23068', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wissmueller-font-liew-cram-schroeder-turner-crossley-mackay-etal-proteinproteininteractionsanalysisofafalsepositivegstpulldownresult-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wissmueller-font-liew-cram-schroeder-turner-crossley-mackay-etal-proteinproteininteractionsanalysisofafalsepositivegstpulldownresult-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Protein-protein interactions: Analysis of a false positive GST pulldown result},\n type = {article},\n year = {2011},\n pages = {2365-2371},\n volume = {79},\n id = {5d9c6e9b-c8ce-3c98-ab37-684192c58e66},\n created = {2023-01-10T01:45:08.736Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:08.736Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {One of the most common ways to demonstrate a direct protein-protein interaction in vitro is the glutathione-S-transferse (GST)-pulldown. Here we report the detailed characterization of a putative interaction between two transcription factor proteins, GATA-1 and Krüppel-like factor 3 (KLF3/BKLF) that show robust interactions in GST-pulldown experiments. Attempts to map the interaction interface of GATA-1 on KLF3 using a mutagenic screening approach did not yield a contiguous binding face on KLF3, suggesting that the interaction might be non-specific. NMR experiments showed that the proteins do not interact at protein concentrations of 50-100 μM. Rather, the GST tag can cause part of KLF3 to misfold. In addition to misfolding, the fact that both proteins are DNA-binding domains appears to introduce binding artifacts (possibly nucleic acid bridging) that cannot be resolved by the addition of nucleases or ethidium bromide (EtBr). This study emphasizes the need for caution in relying on GST-pulldown results and related methods, without convincing confirmation from different approaches. © 2011 Wiley-Liss, Inc.},\n bibtype = {article},\n author = {Wissmueller, S. and Font, J. and Liew, C.W. and Cram, E. and Schroeder, T. and Turner, J. and Crossley, M. and Mackay, J.P. and Matthews, J.M.},\n doi = {10.1002/prot.23068},\n journal = {Proteins: Structure, Function and Bioinformatics},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  One of the most common ways to demonstrate a direct protein-protein interaction in vitro is the glutathione-S-transferse (GST)-pulldown. Here we report the detailed characterization of a putative interaction between two transcription factor proteins, GATA-1 and Krüppel-like factor 3 (KLF3/BKLF) that show robust interactions in GST-pulldown experiments. Attempts to map the interaction interface of GATA-1 on KLF3 using a mutagenic screening approach did not yield a contiguous binding face on KLF3, suggesting that the interaction might be non-specific. NMR experiments showed that the proteins do not interact at protein concentrations of 50-100 μM. Rather, the GST tag can cause part of KLF3 to misfold. In addition to misfolding, the fact that both proteins are DNA-binding domains appears to introduce binding artifacts (possibly nucleic acid bridging) that cannot be resolved by the addition of nucleases or ethidium bromide (EtBr). This study emphasizes the need for caution in relying on GST-pulldown results and related methods, without convincing confirmation from different approaches. © 2011 Wiley-Liss, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gamsjaeger-webb-lamonica-billin-blobel-mackay-structuralbasisandspecificityofacetylatedtranscriptionfactorgata1recognitionbybetfamilybromodomainproteinbrd3-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural basis and specificity of acetylated transcription factor GATA1 recognition by BET family bromodomain protein Brd3.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gamsjaeger,  R.; Webb,  S.; Lamonica,  J.; Billin,  A.; Blobel,  G.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Molecular and Cellular Biology</i>, 31(13): 2632-2640.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1128/MCB.05413-11\"\n     onclick=\"log_download('gamsjaeger-webb-lamonica-billin-blobel-mackay-structuralbasisandspecificityofacetylatedtranscriptionfactorgata1recognitionbybetfamilybromodomainproteinbrd3-2011', 'http://doi.org/10.1128/MCB.05413-11', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gamsjaeger-webb-lamonica-billin-blobel-mackay-structuralbasisandspecificityofacetylatedtranscriptionfactorgata1recognitionbybetfamilybromodomainproteinbrd3-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gamsjaeger-webb-lamonica-billin-blobel-mackay-structuralbasisandspecificityofacetylatedtranscriptionfactorgata1recognitionbybetfamilybromodomainproteinbrd3-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural basis and specificity of acetylated transcription factor GATA1 recognition by BET family bromodomain protein Brd3},\n type = {article},\n year = {2011},\n pages = {2632-2640},\n volume = {31},\n id = {429de600-d97d-3c2c-9f33-97c9b5577dcd},\n created = {2023-01-10T01:45:09.655Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:09.655Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Recent data demonstrate that small synthetic compounds specifically targeting bromodomain proteins can modulate the expression of cancer-related or inflammatory genes. Although these studies have focused on the ability of bromodomains to recognize acetylated histones, it is increasingly becoming clear that histone-like modifications exist on other important proteins, such as transcription factors. However, our understanding of the molecular mechanisms through which these modifications modulate protein function is far from complete. The transcription factor GATA1 can be acetylated at lysine residues adjacent to the zinc finger domains, and this acetylation is essential for the normal chromatin occupancy of GATA1. We have recently identified the bromodomain-containing protein Brd3 as a cofactor that interacts with acetylated GATA1 and shown that this interaction is essential for the targeting of GATA1 to chromatin. Here we describe the structural basis for this interaction. Our data reveal for the first time the molecular details of an interaction between a transcription factor bearing multiple acetylation modifications and its cognate recognition module. We also show that this interaction can be inhibited by an acetyllysine mimic, highlighting the importance of further increasing the specificity of compounds that target bromodomain and extraterminal (BET) bromodomains in order to fully realize their therapeutic potential. © 2011, American Society for Microbiology.},\n bibtype = {article},\n author = {Gamsjaeger, R. and Webb, S.R. and Lamonica, J.M. and Billin, A. and Blobel, G.A. and Mackay, J.P.},\n doi = {10.1128/MCB.05413-11},\n journal = {Molecular and Cellular Biology},\n number = {13}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Recent data demonstrate that small synthetic compounds specifically targeting bromodomain proteins can modulate the expression of cancer-related or inflammatory genes. Although these studies have focused on the ability of bromodomains to recognize acetylated histones, it is increasingly becoming clear that histone-like modifications exist on other important proteins, such as transcription factors. However, our understanding of the molecular mechanisms through which these modifications modulate protein function is far from complete. The transcription factor GATA1 can be acetylated at lysine residues adjacent to the zinc finger domains, and this acetylation is essential for the normal chromatin occupancy of GATA1. We have recently identified the bromodomain-containing protein Brd3 as a cofactor that interacts with acetylated GATA1 and shown that this interaction is essential for the targeting of GATA1 to chromatin. Here we describe the structural basis for this interaction. Our data reveal for the first time the molecular details of an interaction between a transcription factor bearing multiple acetylation modifications and its cognate recognition module. We also show that this interaction can be inhibited by an acetyllysine mimic, highlighting the importance of further increasing the specificity of compounds that target bromodomain and extraterminal (BET) bromodomains in order to fully realize their therapeutic potential. © 2011, American Society for Microbiology.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mansfield-musselman-kwan-oliver-garske-davrazou-denu-kutateladze-etal-planthomeodomainphdfingersofchd4arehistoneh3bindingmoduleswithpreferenceforunmodifiedh3k4andmethylatedh3k9-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Plant homeodomain (PHD) fingers of CHD4 are histone H3-binding modules with preference for unmodified H3K4 and methylated H3K9.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mansfield,  R.; Musselman,  C.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Oliver,  S.; Garske,  A.; Davrazou,  F.; Denu,  J.; Kutateladze,  T.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 286(13): 11779-11791.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M110.208207\"\n     onclick=\"log_download('mansfield-musselman-kwan-oliver-garske-davrazou-denu-kutateladze-etal-planthomeodomainphdfingersofchd4arehistoneh3bindingmoduleswithpreferenceforunmodifiedh3k4andmethylatedh3k9-2011', 'http://doi.org/10.1074/jbc.M110.208207', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mansfield-musselman-kwan-oliver-garske-davrazou-denu-kutateladze-etal-planthomeodomainphdfingersofchd4arehistoneh3bindingmoduleswithpreferenceforunmodifiedh3k4andmethylatedh3k9-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mansfield-musselman-kwan-oliver-garske-davrazou-denu-kutateladze-etal-planthomeodomainphdfingersofchd4arehistoneh3bindingmoduleswithpreferenceforunmodifiedh3k4andmethylatedh3k9-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Plant homeodomain (PHD) fingers of CHD4 are histone H3-binding modules with preference for unmodified H3K4 and methylated H3K9},\n type = {article},\n year = {2011},\n pages = {11779-11791},\n volume = {286},\n id = {5cbc3d31-ec2f-35ec-a0c2-85fb87abd270},\n created = {2023-01-10T01:45:10.576Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:10.576Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {A major challenge in chromatin biology is to understand the mechanisms by which chromatin is remodeled into active or inactive states as required during development and cell differentiation. One complex implicated in these processes is the nucleosome remodeling and histone deacetylase (NuRD) complex, which contains both histone deacetylase and nucleosome remodeling activities and has been implicated in the silencing of subsets of genes involved in various stages of cellular development. Chromodomain-helicase-DNA-binding protein 4 (CHD4) is a core component of the NuRD complex and contains a nucleosome remodeling ATPase domain along with two chromodomains and two plant homeodomain (PHD) fingers. We have previously demonstrated that the second PHD finger of CHD4 binds peptides corresponding to the N terminus of histone H3 methylated at Lys9. Here, we determine the solution structure of PHD2 in complex with H3K9me3, revealing the molecular basis of histone recognition, including a cation-π recognition mechanism for methylated Lys9. Additionally, we demonstrate that the first PHD finger also exhibits binding to the N terminus of H3, and we establish the histone-binding surface of this domain. This is the first instance where histone binding ability has been demonstrated for two separate PHD modules within the one protein. These findings suggest that CHD4 could bind to two H3 N-terminal tails on the same nucleosome or on two separate nucleosomes simultaneously, presenting exciting implications for the mechanism by which CHD4 and the NuRD complex could direct chromatin remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Mansfield, R.E. and Musselman, C.A. and Kwan, A.H. and Oliver, S.S. and Garske, A.L. and Davrazou, F. and Denu, J.M. and Kutateladze, T.G. and Mackay, J.P.},\n doi = {10.1074/jbc.M110.208207},\n journal = {Journal of Biological Chemistry},\n number = {13}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A major challenge in chromatin biology is to understand the mechanisms by which chromatin is remodeled into active or inactive states as required during development and cell differentiation. One complex implicated in these processes is the nucleosome remodeling and histone deacetylase (NuRD) complex, which contains both histone deacetylase and nucleosome remodeling activities and has been implicated in the silencing of subsets of genes involved in various stages of cellular development. Chromodomain-helicase-DNA-binding protein 4 (CHD4) is a core component of the NuRD complex and contains a nucleosome remodeling ATPase domain along with two chromodomains and two plant homeodomain (PHD) fingers. We have previously demonstrated that the second PHD finger of CHD4 binds peptides corresponding to the N terminus of histone H3 methylated at Lys9. Here, we determine the solution structure of PHD2 in complex with H3K9me3, revealing the molecular basis of histone recognition, including a cation-π recognition mechanism for methylated Lys9. Additionally, we demonstrate that the first PHD finger also exhibits binding to the N terminus of H3, and we establish the histone-binding surface of this domain. This is the first instance where histone binding ability has been demonstrated for two separate PHD modules within the one protein. These findings suggest that CHD4 could bind to two H3 N-terminal tails on the same nucleosome or on two separate nucleosomes simultaneously, presenting exciting implications for the mechanism by which CHD4 and the NuRD complex could direct chromatin remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"nguyen-mansfield-leung-vaz-loughlin-grant-mackay-characterizationofafamilyofranbp2typezincfingersthatcanrecognizesinglestrandedrna-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Characterization of a family of RanBP2-Type zinc fingers that can recognize single-stranded RNA.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Nguyen,  C.; Mansfield,  R.; Leung,  W.; Vaz,  P.; Loughlin,  F.; Grant,  R.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 407(2): 273-283.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2010.12.041\"\n     onclick=\"log_download('nguyen-mansfield-leung-vaz-loughlin-grant-mackay-characterizationofafamilyofranbp2typezincfingersthatcanrecognizesinglestrandedrna-2011', 'http://doi.org/10.1016/j.jmb.2010.12.041', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/nguyen-mansfield-leung-vaz-loughlin-grant-mackay-characterizationofafamilyofranbp2typezincfingersthatcanrecognizesinglestrandedrna-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('nguyen-mansfield-leung-vaz-loughlin-grant-mackay-characterizationofafamilyofranbp2typezincfingersthatcanrecognizesinglestrandedrna-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Characterization of a family of RanBP2-Type zinc fingers that can recognize single-stranded RNA},\n type = {article},\n year = {2011},\n pages = {273-283},\n volume = {407},\n id = {48de412f-0ebf-3005-9f78-2ff45c65410e},\n created = {2023-01-10T01:45:11.498Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:11.498Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The recognition of single-stranded RNA (ssRNA) is an important aspect of gene regulation, and a number of different classes of protein domains that recognize ssRNA in a sequence-specific manner have been identified. Recently, we demonstrated that the RanBP2-type zinc finger (ZnF) domains from the human splicing factor ZnF Ran binding domain-containing protein 2 (ZRANB2) can bind to a sequence containing the consensus AGGUAA. Six other human proteins, namely, Ewing&#x27;s sarcoma (EWS), translocated in liposarcoma (TLS)/FUS, RNA-binding protein 56 (RBP56), RNA-binding motif 5 (RBM5), RNA-binding motif 10 (RBM10) and testis-expressed sequence 13A (TEX13A), each contains a single ZnF with homology to the ZRANB2 ZnFs, and several of these proteins have been implicated in the regulation of mRNA processing. Here, we show that all of these ZnFs are able to bind with micromolar affinities to ssRNA containing a GGU motif. NMR titration data reveal that binding is mediated by the corresponding surfaces on each ZnF, and we also show that sequence selectivity is largely limited to the GGU core motif and that substitution of the three flanking adenines that were selected in our original selection experiment has a minimal effect on binding affinity. These data establish a subset of RanBP2-type ZnFs as a new family of ssRNA-binding motifs. © 2011 Elsevier Ltd.},\n bibtype = {article},\n author = {Nguyen, C.D. and Mansfield, R.E. and Leung, W. and Vaz, P.M. and Loughlin, F.E. and Grant, R.P. and MacKay, J.P.},\n doi = {10.1016/j.jmb.2010.12.041},\n journal = {Journal of Molecular Biology},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The recognition of single-stranded RNA (ssRNA) is an important aspect of gene regulation, and a number of different classes of protein domains that recognize ssRNA in a sequence-specific manner have been identified. Recently, we demonstrated that the RanBP2-type zinc finger (ZnF) domains from the human splicing factor ZnF Ran binding domain-containing protein 2 (ZRANB2) can bind to a sequence containing the consensus AGGUAA. Six other human proteins, namely, Ewing's sarcoma (EWS), translocated in liposarcoma (TLS)/FUS, RNA-binding protein 56 (RBP56), RNA-binding motif 5 (RBM5), RNA-binding motif 10 (RBM10) and testis-expressed sequence 13A (TEX13A), each contains a single ZnF with homology to the ZRANB2 ZnFs, and several of these proteins have been implicated in the regulation of mRNA processing. Here, we show that all of these ZnFs are able to bind with micromolar affinities to ssRNA containing a GGU motif. NMR titration data reveal that binding is mediated by the corresponding surfaces on each ZnF, and we also show that sequence selectivity is largely limited to the GGU core motif and that substitution of the three flanking adenines that were selected in our original selection experiment has a minimal effect on binding affinity. These data establish a subset of RanBP2-type ZnFs as a new family of ssRNA-binding motifs. © 2011 Elsevier Ltd.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wilkinson-White,  L.; Gamsjaeger,  R.; Dastmalchi,  S.; Wienert,  B.; Stokes,  P.; Crossley,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 108(35): 14443-14448.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.1105898108\"\n     onclick=\"log_download('wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011', 'http://doi.org/10.1073/pnas.1105898108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wilkinsonwhite-gamsjaeger-dastmalchi-wienert-stokes-crossley-mackay-matthews-structuralbasisofsimultaneousrecruitmentofthetranscriptionalregulatorslmo2andfog1zfpm1bythetranscriptionfactorgata1-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural basis of simultaneous recruitment of the transcriptional regulators LMO2 and FOG1/ZFPM1 by the transcription factor GATA1},\n type = {article},\n year = {2011},\n pages = {14443-14448},\n volume = {108},\n id = {765574bf-c2e8-3a13-8a8c-db663be1661f},\n created = {2023-01-10T01:45:12.413Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:12.413Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (&lt;50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.},\n bibtype = {article},\n author = {Wilkinson-White, L. and Gamsjaeger, R. and Dastmalchi, S. and Wienert, B. and Stokes, P.H. and Crossley, M. and Mackay, J.P. and Matthews, J.M.},\n doi = {10.1073/pnas.1105898108},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {35}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The control of red blood cell and megakaryocyte development by the regulatory protein GATA1 is a paradigm for transcriptional regulation of gene expression in cell lineage differentiation and maturation. Most GATA1-regulated events require GATA1 to bind FOG1, and essentially all GATA1-activated genes are cooccupied by a TAL1/E2A/LMO2/LDB1 complex; however, it is not known whether FOG1 and TAL1/E2A/LMO2/LDB1 are simultaneously recruited by GATA1. Our structural data reveal that the FOG1-binding domain of GATA1, the N finger, can also directly contact LMO2 and show that, despite the small size (<50 residues) of the GATA1 N finger, both FOG1 and LMO2 can simultaneously bind this domain. LMO2 in turn can simultaneously contact both GATA1 and the DNA-binding protein TAL1/E2A at bipartite E-box/WGATAR sites. Taken together, our data provide the first structural snapshot of multiprotein complex formation at GATA1-dependent genes and support a model in which FOG1 and TAL1/E2A/LMO2/LDB1 can cooccupy E-box/WGATAR sites to facilitate GATA1-mediated activation of gene activation.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"bieri-kwan-mobli-king-mackay-gooley-macromolecularnmrspectroscopyforthenonspectroscopistbeyondmacromolecularsolutionstructuredetermination-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Macromolecular NMR spectroscopy for the non-spectroscopist: Beyond macromolecular solution structure determination.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Bieri,  M.; Kwan,  A.; Mobli,  M.; King,  G.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Gooley,  P.\n</span>\n\n  \n\n</span>\n\n  <i>FEBS Journal</i>, 278(5): 704-715.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1111/j.1742-4658.2011.08005.x\"\n     onclick=\"log_download('bieri-kwan-mobli-king-mackay-gooley-macromolecularnmrspectroscopyforthenonspectroscopistbeyondmacromolecularsolutionstructuredetermination-2011', 'http://doi.org/10.1111/j.1742-4658.2011.08005.x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/bieri-kwan-mobli-king-mackay-gooley-macromolecularnmrspectroscopyforthenonspectroscopistbeyondmacromolecularsolutionstructuredetermination-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('bieri-kwan-mobli-king-mackay-gooley-macromolecularnmrspectroscopyforthenonspectroscopistbeyondmacromolecularsolutionstructuredetermination-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Macromolecular NMR spectroscopy for the non-spectroscopist: Beyond macromolecular solution structure determination},\n type = {article},\n year = {2011},\n pages = {704-715},\n volume = {278},\n id = {0c69b0a8-cbe5-3ad1-acb3-27b8003b448f},\n created = {2023-01-10T01:45:13.329Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:13.329Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {A strength of NMR spectroscopy is its ability to monitor, on an atomic level, molecular changes and interactions. In this review, which is intended for non-spectroscopist, we describe major uses of NMR in protein science beyond solution structure determination. After first touching on how NMR can be used to quickly determine whether a mutation induces structural perturbations in a protein, we describe the unparalleled ability of NMR to monitor binding interactions over a wide range of affinities, molecular masses and solution conditions. We discuss the use of NMR to measure the dynamics of proteins at the atomic level and over a wide range of timescales. Finally, we outline new and expanding areas such as macromolecular structure determination in multicomponent systems, as well as in the solid state and in vivo. © 2011 FEBS.},\n bibtype = {article},\n author = {Bieri, M. and Kwan, A.H. and Mobli, M. and King, G.F. and MacKay, J.P. and Gooley, P.R.},\n doi = {10.1111/j.1742-4658.2011.08005.x},\n journal = {FEBS Journal},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A strength of NMR spectroscopy is its ability to monitor, on an atomic level, molecular changes and interactions. In this review, which is intended for non-spectroscopist, we describe major uses of NMR in protein science beyond solution structure determination. After first touching on how NMR can be used to quickly determine whether a mutation induces structural perturbations in a protein, we describe the unparalleled ability of NMR to monitor binding interactions over a wide range of affinities, molecular masses and solution conditions. We discuss the use of NMR to measure the dynamics of proteins at the atomic level and over a wide range of timescales. Finally, we outline new and expanding areas such as macromolecular structure determination in multicomponent systems, as well as in the solid state and in vivo. © 2011 FEBS.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-font-segal-erratumtheprospectsfordesignersinglestrandedrnabindingproteinsnaturestructuralandmolecularbiology201118256261-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Erratum: The prospects for designer single-stranded RNA-binding proteins (Nature Structural and Molecular Biology (2011) 18 (256-261)).\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Font,  J.;  and Segal,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Structural and Molecular Biology</i>, 18(4): 516.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/nsmb0411-516e\"\n     onclick=\"log_download('mackay-font-segal-erratumtheprospectsfordesignersinglestrandedrnabindingproteinsnaturestructuralandmolecularbiology201118256261-2011', 'http://doi.org/10.1038/nsmb0411-516e', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-font-segal-erratumtheprospectsfordesignersinglestrandedrnabindingproteinsnaturestructuralandmolecularbiology201118256261-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-font-segal-erratumtheprospectsfordesignersinglestrandedrnabindingproteinsnaturestructuralandmolecularbiology201118256261-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Erratum: The prospects for designer single-stranded RNA-binding proteins (Nature Structural and Molecular Biology (2011) 18 (256-261))},\n type = {article},\n year = {2011},\n pages = {516},\n volume = {18},\n id = {af45e7f3-95df-3e38-b1e4-265877fed464},\n created = {2023-01-10T01:45:14.370Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:14.370Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {MacKay, J.P. and Font, J. and Segal, D.J.},\n doi = {10.1038/nsmb0411-516e},\n journal = {Nature Structural and Molecular Biology},\n number = {4}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kwan-mobli-gooley-king-mackay-macromolecularnmrspectroscopyforthenonspectroscopist-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Macromolecular NMR spectroscopy for the non-spectroscopist.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Mobli,  M.; Gooley,  P.; King,  G.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>FEBS Journal</i>, 278(5): 687-703.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1111/j.1742-4658.2011.08004.x\"\n     onclick=\"log_download('kwan-mobli-gooley-king-mackay-macromolecularnmrspectroscopyforthenonspectroscopist-2011', 'http://doi.org/10.1111/j.1742-4658.2011.08004.x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kwan-mobli-gooley-king-mackay-macromolecularnmrspectroscopyforthenonspectroscopist-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kwan-mobli-gooley-king-mackay-macromolecularnmrspectroscopyforthenonspectroscopist-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Macromolecular NMR spectroscopy for the non-spectroscopist},\n type = {article},\n year = {2011},\n pages = {687-703},\n volume = {278},\n id = {ed62160d-f565-3f31-ab39-6f55529dd09e},\n created = {2023-01-10T01:45:15.274Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:15.274Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {NMR spectroscopy is a powerful tool for studying the structure, function and dynamics of biological macromolecules. However, non-spectroscopists often find NMR theory daunting and data interpretation nontrivial. As the first of two back-to-back reviews on NMR spectroscopy aimed at non-spectroscopists, the present review first provides an introduction to the basics of macromolecular NMR spectroscopy, including a discussion of typical sample requirements and what information can be obtained from simple NMR experiments. We then review the use of NMR spectroscopy for determining the 3D structures of macromolecules and examine how to judge the quality of NMR-derived structures. © 2011 FEBS.},\n bibtype = {article},\n author = {Kwan, A.H. and Mobli, M. and Gooley, P.R. and King, G.F. and MacKay, J.P.},\n doi = {10.1111/j.1742-4658.2011.08004.x},\n journal = {FEBS Journal},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  NMR spectroscopy is a powerful tool for studying the structure, function and dynamics of biological macromolecules. However, non-spectroscopists often find NMR theory daunting and data interpretation nontrivial. As the first of two back-to-back reviews on NMR spectroscopy aimed at non-spectroscopists, the present review first provides an introduction to the basics of macromolecular NMR spectroscopy, including a discussion of typical sample requirements and what information can be obtained from simple NMR experiments. We then review the use of NMR spectroscopy for determining the 3D structures of macromolecules and examine how to judge the quality of NMR-derived structures. © 2011 FEBS.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"lejon-thong-murthy-alqarni-murzina-blobel-laue-mackay-insightsintoassociationofthenurdcomplexwithfog1fromthecrystalstructureofanrbap48fog1complex-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Insights into association of the NuRD complex with FOG-1 from the crystal structure of an RbAp48·FOG-1 complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Lejon,  S.; Thong,  S.; Murthy,  A.; AlQarni,  S.; Murzina,  N.; Blobel,  G.; Laue,  E.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 286(2): 1196-1203.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M110.195842\"\n     onclick=\"log_download('lejon-thong-murthy-alqarni-murzina-blobel-laue-mackay-insightsintoassociationofthenurdcomplexwithfog1fromthecrystalstructureofanrbap48fog1complex-2011', 'http://doi.org/10.1074/jbc.M110.195842', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/lejon-thong-murthy-alqarni-murzina-blobel-laue-mackay-insightsintoassociationofthenurdcomplexwithfog1fromthecrystalstructureofanrbap48fog1complex-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('lejon-thong-murthy-alqarni-murzina-blobel-laue-mackay-insightsintoassociationofthenurdcomplexwithfog1fromthecrystalstructureofanrbap48fog1complex-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Insights into association of the NuRD complex with FOG-1 from the crystal structure of an RbAp48·FOG-1 complex},\n type = {article},\n year = {2011},\n pages = {1196-1203},\n volume = {286},\n id = {0dcda68f-44aa-33d1-afea-ef433768b02c},\n created = {2023-01-10T01:45:16.197Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:16.197Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Chromatin-modifying complexes such as the NuRD complex are recruited to particular genomic sites by gene-specific nuclear factors. Overall, however, little is known about the molecular basis for these interactions. Here, we present the 1.9 Å resolution crystal structure of the NuRD subunit RbAp48 bound to the 15 N-terminal amino acids of the GATA-1 cofactor FOG-1. The FOG-1 peptide contacts a negatively charged binding pocket on top of the RbAp48 β-propeller that is distinct from the binding surface used by RpAp48 to contact histone H4. We further show that RbAp48 interacts with the NuRD subunit MTA-1 via a surface that is distinct from its FOG-binding pocket, providing a first glimpse into the way in which NuRD assembly facilitates interactions with cofactors. Our RbAp48·FOG-1 structure provides insight into the molecular determinants of FOG-1-dependent association with the NuRD complex and into the links between transcription regulation and nucleosome remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Lejon, S. and Thong, S.Y. and Murthy, A. and AlQarni, S. and Murzina, N.V. and Blobel, G.A. and Laue, E.D. and Mackay, J.P.},\n doi = {10.1074/jbc.M110.195842},\n journal = {Journal of Biological Chemistry},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Chromatin-modifying complexes such as the NuRD complex are recruited to particular genomic sites by gene-specific nuclear factors. Overall, however, little is known about the molecular basis for these interactions. Here, we present the 1.9 Å resolution crystal structure of the NuRD subunit RbAp48 bound to the 15 N-terminal amino acids of the GATA-1 cofactor FOG-1. The FOG-1 peptide contacts a negatively charged binding pocket on top of the RbAp48 β-propeller that is distinct from the binding surface used by RpAp48 to contact histone H4. We further show that RbAp48 interacts with the NuRD subunit MTA-1 via a surface that is distinct from its FOG-binding pocket, providing a first glimpse into the way in which NuRD assembly facilitates interactions with cofactors. Our RbAp48·FOG-1 structure provides insight into the molecular determinants of FOG-1-dependent association with the NuRD complex and into the links between transcription regulation and nucleosome remodeling. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-font-segal-theprospectsfordesignersinglestrandedrnabindingproteins-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The prospects for designer single-stranded RNA-binding proteins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Font,  J.;  and Segal,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Structural and Molecular Biology</i>, 18(3): 256-261.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/nsmb.2005\"\n     onclick=\"log_download('mackay-font-segal-theprospectsfordesignersinglestrandedrnabindingproteins-2011', 'http://doi.org/10.1038/nsmb.2005', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-font-segal-theprospectsfordesignersinglestrandedrnabindingproteins-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-font-segal-theprospectsfordesignersinglestrandedrnabindingproteins-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The prospects for designer single-stranded RNA-binding proteins},\n type = {article},\n year = {2011},\n pages = {256-261},\n volume = {18},\n id = {ff45d397-50a9-36fc-8a33-31782437940d},\n created = {2023-01-10T01:45:17.102Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:17.102Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Spectacular progress has been made in the design of proteins that recognize double-stranded DNA with a chosen specificity, to the point that designer DNA-binding proteins can be ordered commercially. This success raises the question of whether it will be possible to engineer libraries of proteins that can recognize RNA with tailored specificity. Given the recent explosion in the number and diversity of RNA species demonstrated to play roles in biology, designer RNA-binding proteins are set to become valuable tools, both in the research laboratory and potentially in the clinic. Here we discuss the prospects for the realization of this idea. © 2011 Nature America, Inc. All rights reserved.},\n bibtype = {article},\n author = {MacKay, J.P. and Font, J. and Segal, D.J.},\n doi = {10.1038/nsmb.2005},\n journal = {Nature Structural and Molecular Biology},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Spectacular progress has been made in the design of proteins that recognize double-stranded DNA with a chosen specificity, to the point that designer DNA-binding proteins can be ordered commercially. This success raises the question of whether it will be possible to engineer libraries of proteins that can recognize RNA with tailored specificity. Given the recent explosion in the number and diversity of RNA species demonstrated to play roles in biology, designer RNA-binding proteins are set to become valuable tools, both in the research laboratory and potentially in the clinic. Here we discuss the prospects for the realization of this idea. © 2011 Nature America, Inc. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"setiyaputra-mackay-patrick-thestructureofatruncatedphosphoribosylanthranilateisomerasesuggestsaunifiedmodelforevolutionoftheinf8infbarrelfold-2011\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The structure of a truncated phosphoribosylanthranilate isomerase suggests a unified model for evolution of the (βα)<inf>8</inf> barrel fold.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Setiyaputra,  S.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Patrick,  W.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 408(2): 291-303.  2011.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2011.02.048\"\n     onclick=\"log_download('setiyaputra-mackay-patrick-thestructureofatruncatedphosphoribosylanthranilateisomerasesuggestsaunifiedmodelforevolutionoftheinf8infbarrelfold-2011', 'http://doi.org/10.1016/j.jmb.2011.02.048', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/setiyaputra-mackay-patrick-thestructureofatruncatedphosphoribosylanthranilateisomerasesuggestsaunifiedmodelforevolutionoftheinf8infbarrelfold-2011\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('setiyaputra-mackay-patrick-thestructureofatruncatedphosphoribosylanthranilateisomerasesuggestsaunifiedmodelforevolutionoftheinf8infbarrelfold-2011')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The structure of a truncated phosphoribosylanthranilate isomerase suggests a unified model for evolution of the (βα)&lt;inf&gt;8&lt;/inf&gt; barrel fold},\n type = {article},\n year = {2011},\n pages = {291-303},\n volume = {408},\n id = {484786a4-2a38-33ce-8896-5cba612e7ef8},\n created = {2023-01-10T01:45:18.013Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:18.013Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The (βα)8 barrel is one of the most common protein folds, and enzymes with this architecture display a remarkable range of catalytic activities. Many of these functions are associated with ancient metabolic pathways, and phylogenetic reconstructions suggest that the (βα)8 barrel was one of the very first protein folds to emerge. Consequently, there is considerable interest in understanding the evolutionary processes that gave rise to this fold. In particular, much attention has been focused on the plausibility of (βα)8 barrel evolution from homodimers of half barrels. However, we previously isolated a three-quarter-barrel-sized fragment of a (βα)8 barrel, termed truncated phosphoribosylanthranilate isomerase (trPRAI), that is soluble and almost as thermostable as full-length N-(5′-phosphoribosyl) anthranilate isomerase (PRAI). Here, we report the NMR-derived structure of trPRAI. The subdomain is monomeric, is well ordered and adopts a native-like structure in solution. Side chains from strands β1 (Glu3 and Lys5), β2 (Tyr25) and β6 (Lys122) of trPRAI repack to shield the hydrophobic core from the solvent. This result demonstrates that three-quarter barrels were viable intermediates in the evolution of the (βα)8 barrel fold. We propose a unified model for (βα)8 barrel evolution that combines our data, previously published work and plausible scenarios for the emergence of (initially error-prone) genetic systems. In this model, the earliest proto-cells contained diverse pools of part-barrel subdomains. Combinatorial assembly of these subdomains gave rise to many distinct lineages of (βα)8 barrel proteins, that is, our model excludes the possibility that there was a single (βα)8 barrel from which all present examples are descended. © 2011 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Setiyaputra, S. and MacKay, J.P. and Patrick, W.M.},\n doi = {10.1016/j.jmb.2011.02.048},\n journal = {Journal of Molecular Biology},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The (βα)8 barrel is one of the most common protein folds, and enzymes with this architecture display a remarkable range of catalytic activities. Many of these functions are associated with ancient metabolic pathways, and phylogenetic reconstructions suggest that the (βα)8 barrel was one of the very first protein folds to emerge. Consequently, there is considerable interest in understanding the evolutionary processes that gave rise to this fold. In particular, much attention has been focused on the plausibility of (βα)8 barrel evolution from homodimers of half barrels. However, we previously isolated a three-quarter-barrel-sized fragment of a (βα)8 barrel, termed truncated phosphoribosylanthranilate isomerase (trPRAI), that is soluble and almost as thermostable as full-length N-(5′-phosphoribosyl) anthranilate isomerase (PRAI). Here, we report the NMR-derived structure of trPRAI. The subdomain is monomeric, is well ordered and adopts a native-like structure in solution. Side chains from strands β1 (Glu3 and Lys5), β2 (Tyr25) and β6 (Lys122) of trPRAI repack to shield the hydrophobic core from the solvent. This result demonstrates that three-quarter barrels were viable intermediates in the evolution of the (βα)8 barrel fold. We propose a unified model for (βα)8 barrel evolution that combines our data, previously published work and plausible scenarios for the emergence of (initially error-prone) genetic systems. In this model, the earliest proto-cells contained diverse pools of part-barrel subdomains. Combinatorial assembly of these subdomains gave rise to many distinct lineages of (βα)8 barrel proteins, that is, our model excludes the possibility that there was a single (βα)8 barrel from which all present examples are descended. © 2011 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2010\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2010')\"\n      onkeypress=\"toggleGroup('group_2010')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2010</span>\n      <span class=\"bibbase_group_count\">\n        \n        (11)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"font-mackay-methodsinmolecularbiology-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://link.springer.com/10.1007/978-1-60761-753-2_29\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'font-mackay-methodsinmolecularbiology-2010', 'http://link.springer.com/10.1007/978-1-60761-753-2_29', event);\">\n    Beyond DNA: Zinc Finger Domains as RNA-Binding Modules.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Font,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  Volume 649 . Methods in Molecular Biology, pages 479-491.  2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://link.springer.com/10.1007/978-1-60761-753-2_29\"\n     onclick=\"log_download('font-mackay-methodsinmolecularbiology-2010', 'http://link.springer.com/10.1007/978-1-60761-753-2_29', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Methods in Molecular Biology [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1007/978-1-60761-753-2_29\"\n     onclick=\"log_download('font-mackay-methodsinmolecularbiology-2010', 'http://doi.org/10.1007/978-1-60761-753-2_29', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/font-mackay-methodsinmolecularbiology-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('font-mackay-methodsinmolecularbiology-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@inbook{\n type = {inbook},\n year = {2010},\n keywords = {RNA-binding domain,Zinc fingers,protein design,protein interaction domain},\n pages = {479-491},\n volume = {649},\n websites = {http://link.springer.com/10.1007/978-1-60761-753-2_29},\n id = {0f1743b7-0299-3ee3-8152-d528a8b42877},\n created = {2020-12-17T05:29:52.309Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:52.309Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Font2010},\n source_type = {ARTICLE},\n notes = {cited By 21},\n private_publication = {false},\n abstract = {Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.},\n bibtype = {inbook},\n author = {Font, Josep and Mackay, Joel P.},\n doi = {10.1007/978-1-60761-753-2_29},\n chapter = {Beyond DNA: Zinc Finger Domains as RNA-Binding Modules},\n title = {Methods in Molecular Biology}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mansfield-cross-matthews-mackay-aminoacidspeptidesandproteinsinorganicchemistry-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1002/9783527631780.ch12\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'mansfield-cross-matthews-mackay-aminoacidspeptidesandproteinsinorganicchemistry-2010', 'http://doi.wiley.com/10.1002/9783527631780.ch12', event);\">\n    Protein Recognition.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mansfield,  R., E.; Cross,  A., J.; Matthews,  J., M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  Volume 2 . Amino Acids, Peptides and Proteins in Organic Chemistry, pages 505-532. Wiley-VCH Verlag GmbH & Co. KGaA, 12 2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1002/9783527631780.ch12\"\n     onclick=\"log_download('mansfield-cross-matthews-mackay-aminoacidspeptidesandproteinsinorganicchemistry-2010', 'http://doi.wiley.com/10.1002/9783527631780.ch12', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Amino Acids, Peptides and Proteins in Organic Chemistry [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/9783527631780.ch12\"\n     onclick=\"log_download('mansfield-cross-matthews-mackay-aminoacidspeptidesandproteinsinorganicchemistry-2010', 'http://doi.org/10.1002/9783527631780.ch12', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mansfield-cross-matthews-mackay-aminoacidspeptidesandproteinsinorganicchemistry-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mansfield-cross-matthews-mackay-aminoacidspeptidesandproteinsinorganicchemistry-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@inbook{\n type = {inbook},\n year = {2010},\n keywords = {Coupled folding and binding,Hotspots,Post-translational modifications,Protein interfaces,Protein recognition},\n pages = {505-532},\n volume = {2},\n websites = {http://doi.wiley.com/10.1002/9783527631780.ch12},\n month = {12},\n publisher = {Wiley-VCH Verlag GmbH &amp; Co. KGaA},\n day = {28},\n city = {Weinheim, Germany},\n id = {31798292-fed8-39f9-8349-947a9649059a},\n created = {2020-12-17T05:29:54.383Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:54.383Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Mansfield2010},\n source_type = {BOOK},\n notes = {cited By 0},\n private_publication = {false},\n bibtype = {inbook},\n author = {Mansfield, Robyn E. and Cross, Arwen J. and Matthews, Jacqueline M. and Mackay, Joel P.},\n doi = {10.1002/9783527631780.ch12},\n chapter = {Protein Recognition},\n title = {Amino Acids, Peptides and Proteins in Organic Chemistry}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"hynson-kwan-jacques-mackay-trewhella-1h13cand15nbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://link.springer.com/10.1007/s12104-010-9237-6\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'hynson-kwan-jacques-mackay-trewhella-1h13cand15nbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010', 'http://link.springer.com/10.1007/s12104-010-9237-6', event);\">\n    1H, 13C and 15N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Hynson,  R., M., G.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H.</a>; Jacques,  D., A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Trewhella,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 4(2): 167-169. 10 2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://link.springer.com/10.1007/s12104-010-9237-6\"\n     onclick=\"log_download('hynson-kwan-jacques-mackay-trewhella-1h13cand15nbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010', 'http://link.springer.com/10.1007/s12104-010-9237-6', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"1H, 13C and 15N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-010-9237-6\"\n     onclick=\"log_download('hynson-kwan-jacques-mackay-trewhella-1h13cand15nbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010', 'http://doi.org/10.1007/s12104-010-9237-6', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/hynson-kwan-jacques-mackay-trewhella-1h13cand15nbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('hynson-kwan-jacques-mackay-trewhella-1h13cand15nbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {1H, 13C and 15N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis},\n type = {article},\n year = {2010},\n keywords = {Bacillus subtilis,Bacterial signal transduction,Histidine kinase inhibition,KipI},\n pages = {167-169},\n volume = {4},\n websites = {http://link.springer.com/10.1007/s12104-010-9237-6},\n month = {10},\n day = {4},\n id = {48c78b62-9179-35cb-ae9d-83d16d7e9f3b},\n created = {2020-12-17T05:29:56.688Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:56.688Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Hynson2010},\n source_type = {ARTICLE},\n notes = {cited By 2},\n private_publication = {false},\n abstract = {KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.},\n bibtype = {article},\n author = {Hynson, Robert M. G. and Kwan, Ann H. and Jacques, David A. and Mackay, Joel P. and Trewhella, Jill},\n doi = {10.1007/s12104-010-9237-6},\n journal = {Biomolecular NMR Assignments},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1002/prot.22752\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010', 'http://doi.wiley.com/10.1002/prot.22752', event);\">\n    Two-state conformational equilibrium in the Par-4 leucine zipper domain.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Schwalbe,  M.; Dutta,  K.; Libich,  D., S.; Venugopal,  H.; Claridge,  J., K.; Gell,  D., A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Edwards,  P., J., B.;  and Pascal,  S., M.\n</span>\n\n  \n\n</span>\n\n  <i>Proteins: Structure, Function, and Bioinformatics</i>, 78(11): n/a-n/a. 4 2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1002/prot.22752\"\n     onclick=\"log_download('schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010', 'http://doi.wiley.com/10.1002/prot.22752', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Two-state conformational equilibrium in the Par-4 leucine zipper domain [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/prot.22752\"\n     onclick=\"log_download('schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010', 'http://doi.org/10.1002/prot.22752', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Two-state conformational equilibrium in the Par-4 leucine zipper domain},\n type = {article},\n year = {2010},\n keywords = {Circular dichroism,Leucine zipper,Prostate apoptosis response factor 4,Solution NMR spectroscopy},\n pages = {n/a-n/a},\n volume = {78},\n websites = {http://doi.wiley.com/10.1002/prot.22752},\n month = {4},\n day = {15},\n id = {90ab4173-c600-3c15-be8a-4dcbdfc4ed4d},\n created = {2020-12-17T05:29:59.280Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:59.280Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Schwalbe2010},\n source_type = {ARTICLE},\n notes = {cited By 5},\n private_publication = {false},\n abstract = {Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.},\n bibtype = {article},\n author = {Schwalbe, Martin and Dutta, Kaushik and Libich, David S. and Venugopal, Hariprasad and Claridge, Jolyon K. and Gell, David A. and Mackay, Joel P. and Edwards, Patrick J. B. and Pascal, Steven M.},\n doi = {10.1002/prot.22752},\n journal = {Proteins: Structure, Function, and Bioinformatics},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"krishnakumar-dickson-weiss-mackay-gell-ahsphaemoglobinstabilizingproteinstabilizesapohaemoglobininapartiallyfoldedstate-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  AHSP (α-haemoglobin-stabilizing protein) stabilizes apo-α-haemoglobin in a partially folded state.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Krishna Kumar,  K.; Dickson,  C.; Weiss,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Gell,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemical Journal</i>, 432(2): 275-282.  2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1042/BJ20100642\"\n     onclick=\"log_download('krishnakumar-dickson-weiss-mackay-gell-ahsphaemoglobinstabilizingproteinstabilizesapohaemoglobininapartiallyfoldedstate-2010', 'http://doi.org/10.1042/BJ20100642', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/krishnakumar-dickson-weiss-mackay-gell-ahsphaemoglobinstabilizingproteinstabilizesapohaemoglobininapartiallyfoldedstate-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('krishnakumar-dickson-weiss-mackay-gell-ahsphaemoglobinstabilizingproteinstabilizesapohaemoglobininapartiallyfoldedstate-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {AHSP (α-haemoglobin-stabilizing protein) stabilizes apo-α-haemoglobin in a partially folded state},\n type = {article},\n year = {2010},\n pages = {275-282},\n volume = {432},\n id = {cfeb53fe-7617-3568-b0ac-944abaedb834},\n created = {2023-01-10T01:45:18.928Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:18.928Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {To produce functional Hb (haemoglobin), nascent α-globin (αo) and β-globin (βo) chains must each bind a single haem molecule (to form αh and βh) and interact together to form heterodimers.The precise sequence of binding events is unknown, and it has been suggested that additional factors might enhance the efficiency of Hb folding. AHSP (α-haemoglobin-stabilizing protein) has been shown previously to bind αh and regulate redox activity of the haem iron. In the present study, we used a combination of classical and dynamic light scattering and NMR spectroscopy to demonstrate that AHSP forms a heterodimeric complex with αo that inhibits αo aggregation and promotes αo folding in the absence of haem. These findings indicate that AHSP may function as an αo-specific chaperone, and suggest an important role for αo in guiding Hb assembly by stabilizing βo and inhibiting off-pathway self-association of βh. © The Authors Journal compilation © 2010 Biochemical Society.},\n bibtype = {article},\n author = {Krishna Kumar, K. and Dickson, C.F. and Weiss, M.J. and Mackay, J.P. and Gell, D.A.},\n doi = {10.1042/BJ20100642},\n journal = {Biochemical Journal},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  To produce functional Hb (haemoglobin), nascent α-globin (αo) and β-globin (βo) chains must each bind a single haem molecule (to form αh and βh) and interact together to form heterodimers.The precise sequence of binding events is unknown, and it has been suggested that additional factors might enhance the efficiency of Hb folding. AHSP (α-haemoglobin-stabilizing protein) has been shown previously to bind αh and regulate redox activity of the haem iron. In the present study, we used a combination of classical and dynamic light scattering and NMR spectroscopy to demonstrate that AHSP forms a heterodimeric complex with αo that inhibits αo aggregation and promotes αo folding in the absence of haem. These findings indicate that AHSP may function as an αo-specific chaperone, and suggest an important role for αo in guiding Hb assembly by stabilizing βo and inhibiting off-pathway self-association of βh. © The Authors Journal compilation © 2010 Biochemical Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"font-mackay-beyonddnazincfingerdomainsasrnabindingmodules-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Beyond DNA: Zinc finger domains as RNA-binding modules.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Font,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  Volume 649  2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/978-1-60761-753-2_29\"\n     onclick=\"log_download('font-mackay-beyonddnazincfingerdomainsasrnabindingmodules-2010', 'http://doi.org/10.1007/978-1-60761-753-2_29', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/font-mackay-beyonddnazincfingerdomainsasrnabindingmodules-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('font-mackay-beyonddnazincfingerdomainsasrnabindingmodules-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@book{\n title = {Beyond DNA: Zinc finger domains as RNA-binding modules},\n type = {book},\n year = {2010},\n source = {Methods in Molecular Biology},\n pages = {479-491},\n volume = {649},\n id = {c6c74829-cc59-3c5a-bd21-95b1c21c0aac},\n created = {2023-01-10T01:45:19.842Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:19.842Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.},\n bibtype = {book},\n author = {Font, J. and MacKay, J.P.},\n doi = {10.1007/978-1-60761-753-2_29}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Over the last 25 years, we have learned that many structural classes of zinc-binding domains (zinc fingers, ZFs) exist and it has become clear that the molecular functions of these domains are by no means limited to the sequence-specific recognition of double-stranded DNA. For example, ZFs can act as protein recognition or RNA-binding modules, and some domains can exhibit more than one function. In this chapter we describe the progress that has been made in understanding the role of ZF domains as RNA-recognition modules, and we speculate about both the prevalence of such functions and the prospects for creating designer ZFs that target RNA. © 2010 Springer Science+Business Media, LLC.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-segal-preface-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Preface.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Segal,  D.\n</span>\n\n  \n\n</span>\n\n  Volume 649  2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-segal-preface-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-segal-preface-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@book{\n title = {Preface},\n type = {book},\n year = {2010},\n source = {Methods in Molecular Biology},\n volume = {649},\n id = {7ce2996b-3608-389e-b725-b249c989c5b4},\n created = {2023-01-10T01:45:20.744Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:20.744Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {book},\n author = {MacKay, J.P. and Segal, D.J.}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wilkinsonwhite-dastmalchi-kwan-ryan-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo2lim2ldb1lidcomplex-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular Lmo2-LIM2/Ldb1-LID complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wilkinson-White,  L.; Dastmalchi,  S.; Kwan,  A.; Ryan,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 4(2): 203-206.  2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-010-9240-y\"\n     onclick=\"log_download('wilkinsonwhite-dastmalchi-kwan-ryan-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo2lim2ldb1lidcomplex-2010', 'http://doi.org/10.1007/s12104-010-9240-y', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wilkinsonwhite-dastmalchi-kwan-ryan-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo2lim2ldb1lidcomplex-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wilkinsonwhite-dastmalchi-kwan-ryan-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlmo2lim2ldb1lidcomplex-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {&lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;15&lt;/sup&gt;N and &lt;sup&gt;13&lt;/sup&gt;C assignments of an intramolecular Lmo2-LIM2/Ldb1-LID complex},\n type = {article},\n year = {2010},\n pages = {203-206},\n volume = {4},\n id = {d3067811-3bab-3f57-99ac-522a48b4c9b3},\n created = {2023-01-10T01:45:21.656Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:21.656Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Lmo2 is a LIM-only protein involved in hematopoiesis and the development of T-cell acute lymphoblastic leukaemia. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the C-terminal LIM domain from Lmo2 tethered to the LIM interaction domain (LID) from LIM domain binding protein 1 (Ldb1). © 2010 Springer Science+Business Media B.V.},\n bibtype = {article},\n author = {Wilkinson-White, L.E. and Dastmalchi, S. and Kwan, A.H. and Ryan, D.P. and MacKay, J.P. and Matthews, J.M.},\n doi = {10.1007/s12104-010-9240-y},\n journal = {Biomolecular NMR Assignments},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Lmo2 is a LIM-only protein involved in hematopoiesis and the development of T-cell acute lymphoblastic leukaemia. Here we report backbone and side chain NMR assignments for an engineered intramolecular complex of the C-terminal LIM domain from Lmo2 tethered to the LIM interaction domain (LID) from LIM domain binding protein 1 (Ldb1). © 2010 Springer Science+Business Media B.V.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Two-state conformational equilibrium in the Par-4 leucine zipper domain.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Schwalbe,  M.; Dutta,  K.; Libich,  D.; Venugopal,  H.; Claridge,  J.; Gell,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Edwards,  P.;  and Pascal,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Proteins: Structure, Function and Bioinformatics</i>, 78(11): 2433-2449.  2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/prot.22752\"\n     onclick=\"log_download('schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010', 'http://doi.org/10.1002/prot.22752', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('schwalbe-dutta-libich-venugopal-claridge-gell-mackay-edwards-etal-twostateconformationalequilibriuminthepar4leucinezipperdomain-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Two-state conformational equilibrium in the Par-4 leucine zipper domain},\n type = {article},\n year = {2010},\n pages = {2433-2449},\n volume = {78},\n id = {f7152d3f-03a8-3aab-8417-8bbf484d2488},\n created = {2023-01-10T01:45:22.571Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:22.571Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.},\n bibtype = {article},\n author = {Schwalbe, M. and Dutta, K. and Libich, D.S. and Venugopal, H. and Claridge, J.K. and Gell, D.A. and Mackay, J.P. and Edwards, P.J.B. and Pascal, S.M.},\n doi = {10.1002/prot.22752},\n journal = {Proteins: Structure, Function and Bioinformatics},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium. © 2010 Wiley-Liss, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"hynson-kwan-jacques-mackay-trewhella-sup1suphsup13supcandsup15supnbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Hynson,  R.; Kwan,  A.; Jacques,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Trewhella,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Biomolecular NMR Assignments</i>, 4(2): 167-169.  2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s12104-010-9237-6\"\n     onclick=\"log_download('hynson-kwan-jacques-mackay-trewhella-sup1suphsup13supcandsup15supnbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010', 'http://doi.org/10.1007/s12104-010-9237-6', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/hynson-kwan-jacques-mackay-trewhella-sup1suphsup13supcandsup15supnbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('hynson-kwan-jacques-mackay-trewhella-sup1suphsup13supcandsup15supnbackboneandsidechainresonanceassignmentsofthenterminaldomainofthehistidinekinaseinhibitorkipifrombacillussubtilis-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {&lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;13&lt;/sup&gt;C and &lt;sup&gt;15&lt;/sup&gt;N backbone and side chain resonance assignments of the N-terminal domain of the histidine kinase inhibitor KipI from Bacillus subtilis},\n type = {article},\n year = {2010},\n pages = {167-169},\n volume = {4},\n id = {032a0fa0-83ef-3610-bdcc-94e56a23bb75},\n created = {2023-01-10T01:45:23.480Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:23.480Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.},\n bibtype = {article},\n author = {Hynson, R.M.G. and Kwan, A.H. and Jacques, D.A. and MacKay, J.P. and Trewhella, J.},\n doi = {10.1007/s12104-010-9237-6},\n journal = {Biomolecular NMR Assignments},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  KipI is a sporulation inhibitor in Bacillus subtilis which acts by binding to the dimerisation and histidine phosphotransfer (DHp) domain of KinA, the principle input kinase in the phosphorelay responsible for sporulation. The 15N, 13C and 1H chemical shift assignments of the N-terminal domain of KipI were determined using multidimensional, multinuclear NMR experiments. The N-terminal domain has two conformers and resonance assignments have been made for both conformers. © 2010 Springer Science+Business Media B.V.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mansfield-cross-matthews-mackay-proteinrecognition-2010\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Protein Recognition.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Mansfield,  R.; Cross,  A.; Matthews,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  Volume 2  2010.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/9783527631780.ch12\"\n     onclick=\"log_download('mansfield-cross-matthews-mackay-proteinrecognition-2010', 'http://doi.org/10.1002/9783527631780.ch12', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mansfield-cross-matthews-mackay-proteinrecognition-2010\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mansfield-cross-matthews-mackay-proteinrecognition-2010')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@book{\n title = {Protein Recognition},\n type = {book},\n year = {2010},\n source = {Amino Acids, Peptides and Proteins in Organic Chemistry},\n pages = {505-532},\n volume = {2},\n id = {cf609a26-bd7d-39ca-a12c-7a19ab224d44},\n created = {2023-01-10T01:45:24.389Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:24.389Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {book},\n author = {Mansfield, R.E. and Cross, A.J. and Matthews, J.M. and Mackay, J.P.},\n doi = {10.1002/9783527631780.ch12}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2009\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2009')\"\n      onkeypress=\"toggleGroup('group_2009')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2009</span>\n      <span class=\"bibbase_group_count\">\n        \n        (11)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009', 'https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is', event);\">\n    Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Musselman,  C., A.; Mansfield,  R., E.; Garske,  A., L.; Davrazou,  F.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H.</a>; Oliver,  S., S.; O'Leary,  H.; Denu,  J., M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Kutateladze,  T., G.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemical Journal</i>, 423(2): 179-187. 10 2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is\"\n     onclick=\"log_download('musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009', 'https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1042/BJ20090870\"\n     onclick=\"log_download('musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009', 'http://doi.org/10.1042/BJ20090870', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications},\n type = {article},\n year = {2009},\n keywords = {Chromodomain helicase DNA-binding protein 4 (CHD4),Histone,Methylation,Plant homeodomain (PHD)},\n pages = {179-187},\n volume = {423},\n websites = {https://portlandpress.com/biochemj/article/423/2/179/44685/Binding-of-the-CHD4-PHD2-finger-to-histone-H3-is},\n month = {10},\n day = {15},\n id = {4c2da44b-b1f6-3dcb-a479-0d561aabb87c},\n created = {2020-12-17T05:29:54.740Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:54.740Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Musselman2009},\n source_type = {ARTICLE},\n notes = {cited By 73},\n private_publication = {false},\n abstract = {CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation or methylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2–H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin.},\n bibtype = {article},\n author = {Musselman, Catherine A. and Mansfield, Robyn E. and Garske, Adam L. and Davrazou, Foteini and Kwan, Ann H. and Oliver, Samuel S. and O&#x27;Leary, Heather and Denu, John M. and Mackay, Joel P. and Kutateladze, Tatiana G.},\n doi = {10.1042/BJ20090870},\n journal = {Biochemical Journal},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation or methylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2–H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"oconnell-gamsjaeger-mackay-thestructuralanalysisofprotein128147proteininteractionsbynmrspectroscopy-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://doi.wiley.com/10.1002/pmic.200900303\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'oconnell-gamsjaeger-mackay-thestructuralanalysisofprotein128147proteininteractionsbynmrspectroscopy-2009', 'http://doi.wiley.com/10.1002/pmic.200900303', event);\">\n    The structural analysis of proteinâ&#128;&#147;protein interactions by NMR spectroscopy.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  O'Connell,  M., R.; Gamsjaeger,  R.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>PROTEOMICS</i>, 9(23): 5224-5232. 12 2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://doi.wiley.com/10.1002/pmic.200900303\"\n     onclick=\"log_download('oconnell-gamsjaeger-mackay-thestructuralanalysisofprotein128147proteininteractionsbynmrspectroscopy-2009', 'http://doi.wiley.com/10.1002/pmic.200900303', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"The structural analysis of proteinâ&amp;#128;&amp;#147;protein interactions by NMR spectroscopy [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pmic.200900303\"\n     onclick=\"log_download('oconnell-gamsjaeger-mackay-thestructuralanalysisofprotein128147proteininteractionsbynmrspectroscopy-2009', 'http://doi.org/10.1002/pmic.200900303', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/oconnell-gamsjaeger-mackay-thestructuralanalysisofprotein128147proteininteractionsbynmrspectroscopy-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('oconnell-gamsjaeger-mackay-thestructuralanalysisofprotein128147proteininteractionsbynmrspectroscopy-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The structural analysis of proteinâ&amp;#128;&amp;#147;protein interactions by NMR spectroscopy},\n type = {article},\n year = {2009},\n keywords = {NMR spectroscopy,Protein-protein interactions,Structural biology,Technology},\n pages = {5224-5232},\n volume = {9},\n websites = {http://doi.wiley.com/10.1002/pmic.200900303},\n month = {12},\n id = {fc390463-14cb-327f-93bf-fbec6b3cd8fd},\n created = {2020-12-17T05:29:55.049Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:55.049Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {OConnell2009},\n source_type = {ARTICLE},\n notes = {cited By 44},\n private_publication = {false},\n abstract = {A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH &amp; Co. KGaA.},\n bibtype = {article},\n author = {O&#x27;Connell, Mitchell R. and Gamsjaeger, Roland and Mackay, Joel P.},\n doi = {10.1002/pmic.200900303},\n journal = {PROTEOMICS},\n number = {23}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"oconnell-gamsjaeger-mackay-thestructuralanalysisofproteinproteininteractionsbynmrspectroscopy-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The structural analysis of protein-protein interactions by NMR spectroscopy.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  O'Connell,  M.; Gamsjaeger,  R.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Proteomics</i>, 9(23): 5224-5232.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/pmic.200900303\"\n     onclick=\"log_download('oconnell-gamsjaeger-mackay-thestructuralanalysisofproteinproteininteractionsbynmrspectroscopy-2009', 'http://doi.org/10.1002/pmic.200900303', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/oconnell-gamsjaeger-mackay-thestructuralanalysisofproteinproteininteractionsbynmrspectroscopy-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('oconnell-gamsjaeger-mackay-thestructuralanalysisofproteinproteininteractionsbynmrspectroscopy-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The structural analysis of protein-protein interactions by NMR spectroscopy},\n type = {article},\n year = {2009},\n pages = {5224-5232},\n volume = {9},\n id = {388e096a-15bd-345c-96eb-0a1702c4a9b6},\n created = {2023-01-10T01:45:25.437Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:25.437Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH &amp; Co. KGaA.},\n bibtype = {article},\n author = {O&#x27;Connell, M.R. and Gamsjaeger, R. and Mackay, J.P.},\n doi = {10.1002/pmic.200900303},\n journal = {Proteomics},\n number = {23}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A comprehensive understanding of protein-protein interactions is an important next step in our quest to understand how the information contained in a genome is put into action. Although a number of experimental techniques can report on the existence of a protein-protein interaction, very few can provide detailed structural information. NMR spectroscopy is one of these, and in recent years several complementary NMR approaches, including residual dipolar couplings and the use of paramagnetic effects, have been developed that can provide insight into the structure of protein-protein complexes. In this article, we review these approaches and comment on their strengths and weaknesses. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"macindoe-glockner-vukain-stennard-costa-harvey-mackay-sunde-conformationalstabilityanddnabindingspecificityofthecardiactboxtranscriptionfactortbx20-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Conformational Stability and DNA Binding Specificity of the Cardiac T-Box Transcription Factor Tbx20.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Macindoe,  I.; Glockner,  L.; Vukašin,  P.; Stennard,  F.; Costa,  M.; Harvey,  R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Sunde,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 389(3): 606-618.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2009.04.056\"\n     onclick=\"log_download('macindoe-glockner-vukain-stennard-costa-harvey-mackay-sunde-conformationalstabilityanddnabindingspecificityofthecardiactboxtranscriptionfactortbx20-2009', 'http://doi.org/10.1016/j.jmb.2009.04.056', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/macindoe-glockner-vukain-stennard-costa-harvey-mackay-sunde-conformationalstabilityanddnabindingspecificityofthecardiactboxtranscriptionfactortbx20-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('macindoe-glockner-vukain-stennard-costa-harvey-mackay-sunde-conformationalstabilityanddnabindingspecificityofthecardiactboxtranscriptionfactortbx20-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Conformational Stability and DNA Binding Specificity of the Cardiac T-Box Transcription Factor Tbx20},\n type = {article},\n year = {2009},\n pages = {606-618},\n volume = {389},\n id = {c62d0b72-ffae-3a09-8c2e-63a5222cc5a2},\n created = {2023-01-10T01:45:26.344Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:26.344Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The transcription factor Tbx20 acts within a hierarchy of T-box factors in lineage specification and morphogenesis in the mammalian heart and is mutated in congenital heart disease. T-box family members share a ∼ 20-kDa DNA-binding domain termed the T-box. The question of how highly homologous T-box proteins achieve differential transcriptional control in heart development, while apparently binding to the same DNA sequence, remains unresolved. Here we show that the optimal DNA recognition sequence for the T-box of Tbx20 corresponds to a T-half-site. Furthermore, we demonstrate using purified recombinant domains that distinct T-boxes show significant differences in the affinity and kinetics of binding and in conformational stability, with the T-box of Tbx20 displaying molten globule character. Our data highlight unique features of Tbx20 and suggest mechanistic ways in which cardiac T-box factors might interact synergistically and/or competitively within the cardiac regulatory network. © 2009 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Macindoe, I. and Glockner, L. and Vukašin, P. and Stennard, F.A. and Costa, M.W. and Harvey, R.P. and Mackay, J.P. and Sunde, M.},\n doi = {10.1016/j.jmb.2009.04.056},\n journal = {Journal of Molecular Biology},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The transcription factor Tbx20 acts within a hierarchy of T-box factors in lineage specification and morphogenesis in the mammalian heart and is mutated in congenital heart disease. T-box family members share a ∼ 20-kDa DNA-binding domain termed the T-box. The question of how highly homologous T-box proteins achieve differential transcriptional control in heart development, while apparently binding to the same DNA sequence, remains unresolved. Here we show that the optimal DNA recognition sequence for the T-box of Tbx20 corresponds to a T-half-site. Furthermore, we demonstrate using purified recombinant domains that distinct T-boxes show significant differences in the affinity and kinetics of binding and in conformational stability, with the T-box of Tbx20 displaying molten globule character. Our data highlight unique features of Tbx20 and suggest mechanistic ways in which cardiac T-box factors might interact synergistically and/or competitively within the cardiac regulatory network. © 2009 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gell-feng-zhou-jeffrey-bendak-gow-weiss-shi-etal-acisprolineinhemoglobinstabilizingproteindirectsthestructuralreorganizationofhemoglobin-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gell,  D.; Feng,  L.; Zhou,  S.; Jeffrey,  P.; Bendak,  K.; Gow,  A.; Weiss,  M.; Shi,  Y.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 284(43): 29462-29469.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M109.027045\"\n     onclick=\"log_download('gell-feng-zhou-jeffrey-bendak-gow-weiss-shi-etal-acisprolineinhemoglobinstabilizingproteindirectsthestructuralreorganizationofhemoglobin-2009', 'http://doi.org/10.1074/jbc.M109.027045', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gell-feng-zhou-jeffrey-bendak-gow-weiss-shi-etal-acisprolineinhemoglobinstabilizingproteindirectsthestructuralreorganizationofhemoglobin-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gell-feng-zhou-jeffrey-bendak-gow-weiss-shi-etal-acisprolineinhemoglobinstabilizingproteindirectsthestructuralreorganizationofhemoglobin-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A cis-proline in α-hemoglobin stabilizing protein directs the structural reorganization of α-hemoglobin},\n type = {article},\n year = {2009},\n pages = {29462-29469},\n volume = {284},\n id = {a280cdd1-9f7b-37f1-8e93-e668c33b7b99},\n created = {2023-01-10T01:45:27.256Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:27.256Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {α-Hemoglobin (αHb) stabilizing protein (AHSP) is expressed in erythropoietic tissues as an accessory factor in hemoglobin synthesis. AHSP forms a specific complex with αHb and suppresses the heme-catalyzed evolution of reactive oxygen species by converting αHb to a conformation in which the heme is coordinated at both axial positions by histidine side chains (bis-histidyl coordination). Currently, the detailed mechanism by which AHSP induces structural changes in αHb has not been determined. Here, we present x-ray crystallography, NMR spectroscopy, and mutagenesis data that identify, for the first time, the importance of an evolutionarily conserved proline, Pro30, in loop 1 of AHSP. Mutation of Pro30 to a variety of residue types results in reduced ability to convert αHb. In complex with-Hb, AHSP Pro30 adopts a cis-peptidyl conformation and makes contact with the N terminus of helix G in αHb. Mutations that stabilize the cis-peptidyl conformation of free AHSP, also enhance the αHb conversion activity. These findings suggest that AHSP loop 1 can transmit structural changes to the heme pocket of αHb, and, more generally, highlight the importance of cis-peptidyl prolyl residues in defining the conformation of regulatory protein loops. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Gell, D.A. and Feng, L. and Zhou, S. and Jeffrey, P.D. and Bendak, K. and Gow, A. and Weiss, M.J. and Shi, Y. and Mackay, J.P.},\n doi = {10.1074/jbc.M109.027045},\n journal = {Journal of Biological Chemistry},\n number = {43}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  α-Hemoglobin (αHb) stabilizing protein (AHSP) is expressed in erythropoietic tissues as an accessory factor in hemoglobin synthesis. AHSP forms a specific complex with αHb and suppresses the heme-catalyzed evolution of reactive oxygen species by converting αHb to a conformation in which the heme is coordinated at both axial positions by histidine side chains (bis-histidyl coordination). Currently, the detailed mechanism by which AHSP induces structural changes in αHb has not been determined. Here, we present x-ray crystallography, NMR spectroscopy, and mutagenesis data that identify, for the first time, the importance of an evolutionarily conserved proline, Pro30, in loop 1 of AHSP. Mutation of Pro30 to a variety of residue types results in reduced ability to convert αHb. In complex with-Hb, AHSP Pro30 adopts a cis-peptidyl conformation and makes contact with the N terminus of helix G in αHb. Mutations that stabilize the cis-peptidyl conformation of free AHSP, also enhance the αHb conversion activity. These findings suggest that AHSP loop 1 can transmit structural changes to the heme pocket of αHb, and, more generally, highlight the importance of cis-peptidyl prolyl residues in defining the conformation of regulatory protein loops. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Musselman,  C.; Mansfield,  R.; Garske,  A.; Davrazou,  F.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Oliver,  S.; O'Leary,  H.; Denu,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Kutateladze,  T.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemical Journal</i>, 423(2): 179-187.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1042/BJ20090870\"\n     onclick=\"log_download('musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009', 'http://doi.org/10.1042/BJ20090870', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('musselman-mansfield-garske-davrazou-kwan-oliver-oleary-denu-etal-bindingofthechd4phd2fingertohistoneh3ismodulatedbycovalentmodifications-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications},\n type = {article},\n year = {2009},\n pages = {179-187},\n volume = {423},\n id = {852e680d-f3c3-3677-a4d0-54c4e5f30958},\n created = {2023-01-10T01:45:28.180Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:28.180Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation ormethylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2-H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin. © The Authors Journal compilation. © 2009 Biochemical Society.},\n bibtype = {article},\n author = {Musselman, C.A. and Mansfield, R.E. and Garske, A.L. and Davrazou, F. and Kwan, A.H. and Oliver, S.S. and O&#x27;Leary, H. and Denu, J.M. and Mackay, J.P. and Kutateladze, T.G.},\n doi = {10.1042/BJ20090870},\n journal = {Biochemical Journal},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation ormethylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 μM binding affinity toward unmodified H3 rises to 0.6 μM for H3K9ac and to 0.9 μM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2-H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin. © The Authors Journal compilation. © 2009 Biochemical Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"langley-shojaei-chan-lok-mackay-traut-guss-christopherson-erratumstructureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparumbiochemistry2009481125702570101021bi900043p-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Erratum: Structure and inhibition of orotidine 5#-Monophosphate decarboxylase from plasmodium falciparum (Biochemistry (2009) 48:11 (2570-2570) 10.1021/bi900043p).\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Langley,  D.; Shojaei,  M.; Chan,  C.; Lok,  H.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Traut,  T.; Guss,  J.;  and Christopherson,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 48(11): 2570.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi900043p\"\n     onclick=\"log_download('langley-shojaei-chan-lok-mackay-traut-guss-christopherson-erratumstructureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparumbiochemistry2009481125702570101021bi900043p-2009', 'http://doi.org/10.1021/bi900043p', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/langley-shojaei-chan-lok-mackay-traut-guss-christopherson-erratumstructureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparumbiochemistry2009481125702570101021bi900043p-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('langley-shojaei-chan-lok-mackay-traut-guss-christopherson-erratumstructureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparumbiochemistry2009481125702570101021bi900043p-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Erratum: Structure and inhibition of orotidine 5#-Monophosphate decarboxylase from plasmodium falciparum (Biochemistry (2009) 48:11 (2570-2570) 10.1021/bi900043p)},\n type = {article},\n year = {2009},\n pages = {2570},\n volume = {48},\n id = {600819d9-be33-3029-9db3-d7575a665191},\n created = {2023-01-10T01:45:29.095Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:29.095Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Langley, D.B. and Shojaei, M. and Chan, C. and Lok, H.C. and Mackay, J.P. and Traut, T.W. and Guss, J.M. and Christopherson, R.I.},\n doi = {10.1021/bi900043p},\n journal = {Biochemistry},\n number = {11}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"talib-beck-urathamakul-nguyen-aldrichwright-mackay-ralph-amassspectrometricinvestigationoftheabilityofmetalcomplexestomodulatetranscriptionfactoractivity-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A mass spectrometric investigation of the ability of metal complexes to modulate transcription factor activity.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Talib,  J.; Beck,  J.; Urathamakul,  T.; Nguyen,  C.; Aldrich-Wright,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Ralph,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Chemical Communications</i>, (37): 5546-5548.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1039/b904751d\"\n     onclick=\"log_download('talib-beck-urathamakul-nguyen-aldrichwright-mackay-ralph-amassspectrometricinvestigationoftheabilityofmetalcomplexestomodulatetranscriptionfactoractivity-2009', 'http://doi.org/10.1039/b904751d', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/talib-beck-urathamakul-nguyen-aldrichwright-mackay-ralph-amassspectrometricinvestigationoftheabilityofmetalcomplexestomodulatetranscriptionfactoractivity-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('talib-beck-urathamakul-nguyen-aldrichwright-mackay-ralph-amassspectrometricinvestigationoftheabilityofmetalcomplexestomodulatetranscriptionfactoractivity-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A mass spectrometric investigation of the ability of metal complexes to modulate transcription factor activity},\n type = {article},\n year = {2009},\n pages = {5546-5548},\n id = {674f52e4-6615-311d-9893-ba1bd7e3946c},\n created = {2023-01-10T01:45:30.041Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:30.041Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {ESI mass spectrometry was used to assess the ability of metal complexes to inhibit binding of a transcription factor to a DNA molecule containing its recognition sequence. © 2009 The Royal Society of Chemistry.},\n bibtype = {article},\n author = {Talib, J. and Beck, J.L. and Urathamakul, T. and Nguyen, C.D. and Aldrich-Wright, J.R. and MacKay, J.P. and Ralph, S.F.},\n doi = {10.1039/b904751d},\n journal = {Chemical Communications},\n number = {37}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  ESI mass spectrometry was used to assess the ability of metal complexes to inhibit binding of a transcription factor to a DNA molecule containing its recognition sequence. © 2009 The Royal Society of Chemistry.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"lowry-gamsjaeger-thong-hung-kwan-broitmanmaduro-matthews-maduro-etal-structuralanalysisofmed1revealsunexpecteddiversityinthemechanismofdnarecognitionbygatatypezincfingerdomains-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural analysis of MED-1 reveals unexpected diversity in the mechanism of DNA recognition by GATA-type zinc finger domains.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Lowry,  J.; Gamsjaeger,  R.; Thong,  S.; Hung,  W.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Broitman-Maduro,  G.; Matthews,  J.; Maduro,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 284(9): 5827-5835.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M808712200\"\n     onclick=\"log_download('lowry-gamsjaeger-thong-hung-kwan-broitmanmaduro-matthews-maduro-etal-structuralanalysisofmed1revealsunexpecteddiversityinthemechanismofdnarecognitionbygatatypezincfingerdomains-2009', 'http://doi.org/10.1074/jbc.M808712200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/lowry-gamsjaeger-thong-hung-kwan-broitmanmaduro-matthews-maduro-etal-structuralanalysisofmed1revealsunexpecteddiversityinthemechanismofdnarecognitionbygatatypezincfingerdomains-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('lowry-gamsjaeger-thong-hung-kwan-broitmanmaduro-matthews-maduro-etal-structuralanalysisofmed1revealsunexpecteddiversityinthemechanismofdnarecognitionbygatatypezincfingerdomains-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural analysis of MED-1 reveals unexpected diversity in the mechanism of DNA recognition by GATA-type zinc finger domains},\n type = {article},\n year = {2009},\n pages = {5827-5835},\n volume = {284},\n id = {71df5cc8-308b-387c-9226-824ae9beba6f},\n created = {2023-01-10T01:45:30.973Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:30.973Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {MED-1 is a member of a group of divergent GATA-type zinc finger proteins recently identified in several species of Caenorhabditis. The med genes are transcriptional regulators that are involved in the specification of the mesoderm and endoderm precursor cells in nematodes. Unlike other GATA-type zinc fingers that recognize the consensus sequence (A/C/ T)GATA(A/G), the MED-1 zinc finger (MED1zf) binds the larger and atypical site GTATACT(T/C)3. We have examined the basis for this unusual DNA specificity using a range of biochemical and biophysical approaches. Most strikingly, we show that although the core of the MED1zf structure is similar to that of GATA-1, the basic tail C-terminal to the zinc finger unexpectedly adopts an α-helical structure upon binding DNA. This additional helix appears to contact the major groove of the DNA, making contacts that explain the extended DNA consensus sequence observed for MED1zf. Our data expand the versatility of DNA recognition by GATA-type zinc fingers and perhaps shed new light on the DNA-binding properties of mammalian GATA factors. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Lowry, J.A. and Gamsjaeger, R. and Thong, S.Y. and Hung, W. and Kwan, A.H. and Broitman-Maduro, G. and Matthews, J.M. and Maduro, M. and Mackay, J.P.},\n doi = {10.1074/jbc.M808712200},\n journal = {Journal of Biological Chemistry},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  MED-1 is a member of a group of divergent GATA-type zinc finger proteins recently identified in several species of Caenorhabditis. The med genes are transcriptional regulators that are involved in the specification of the mesoderm and endoderm precursor cells in nematodes. Unlike other GATA-type zinc fingers that recognize the consensus sequence (A/C/ T)GATA(A/G), the MED-1 zinc finger (MED1zf) binds the larger and atypical site GTATACT(T/C)3. We have examined the basis for this unusual DNA specificity using a range of biochemical and biophysical approaches. Most strikingly, we show that although the core of the MED1zf structure is similar to that of GATA-1, the basic tail C-terminal to the zinc finger unexpectedly adopts an α-helical structure upon binding DNA. This additional helix appears to contact the major groove of the DNA, making contacts that explain the extended DNA consensus sequence observed for MED1zf. Our data expand the versatility of DNA recognition by GATA-type zinc fingers and perhaps shed new light on the DNA-binding properties of mammalian GATA factors. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"loughlin-mansfield-vaz-mcgrath-setiyaputra-gamsjaeger-chen-morris-etal-thezincfingersofthesrlikeproteinzranb2aresinglestrandedrnabindingdomainsthatrecognize5splicesitelikesequences-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The zinc fingers of the SR-like protein ZRANB2 are single-stranded RNA-binding domains that recognize 5' splice site-like sequences.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Loughlin,  F.; Mansfield,  R.; Vaz,  P.; McGrath,  A.; Setiyaputra,  S.; Gamsjaeger,  R.; Chen,  E.; Morris,  B.; Guss,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 106(14): 5581-5586.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.0802466106\"\n     onclick=\"log_download('loughlin-mansfield-vaz-mcgrath-setiyaputra-gamsjaeger-chen-morris-etal-thezincfingersofthesrlikeproteinzranb2aresinglestrandedrnabindingdomainsthatrecognize5splicesitelikesequences-2009', 'http://doi.org/10.1073/pnas.0802466106', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/loughlin-mansfield-vaz-mcgrath-setiyaputra-gamsjaeger-chen-morris-etal-thezincfingersofthesrlikeproteinzranb2aresinglestrandedrnabindingdomainsthatrecognize5splicesitelikesequences-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('loughlin-mansfield-vaz-mcgrath-setiyaputra-gamsjaeger-chen-morris-etal-thezincfingersofthesrlikeproteinzranb2aresinglestrandedrnabindingdomainsthatrecognize5splicesitelikesequences-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The zinc fingers of the SR-like protein ZRANB2 are single-stranded RNA-binding domains that recognize 5&#x27; splice site-like sequences},\n type = {article},\n year = {2009},\n pages = {5581-5586},\n volume = {106},\n id = {bbb64689-b326-3416-95e2-8b6b88f2d5e6},\n created = {2023-01-10T01:45:31.914Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:31.914Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The alternative splicing of mRNA is a critical process in higher eukaryotes that generates substantial proteomic diversity. Many of the proteins that are essential to this process contain arginine/serine-rich (RS) domains. ZRANB2 is a widely-expressed and highly-conserved RS-domain protein that can regulate alternative splicing but lacks canonical RNA-binding domains. Instead, it contains 2 RanBP2-type zinc finger (ZnF) domains. We demonstrate that these ZnFs recognize ssRNA with high affinity and specificity. Each ZnF binds to a single AGGUAA motif and the 2 domains combine to recognize AGGUAA (Nx) AGGUAA double sites, suggesting that ZRANB2 regulates alternative splicing via a direct interaction with pre-mRNA at sites that resemble the consensus 5&#x27; splice site. We show using X-ray crystallography that recognition of an AGGUAA motif by a single ZnF is dominated by side-chain hydrogen bonds to the bases and formation of a guanine-tryptophan-guanine &#x27;&#x27;ladder.&#x27;&#x27; A number of other human proteins that function in RNA processing also contain RanBP2 ZnFs in which the RNA-binding residues of ZRANB2 are conserved. The ZnFs of ZRANB2 therefore define another class of RNA-binding domain, advancing our understanding of RNA recognition and emphasizing the versatility of domains in molecular recognition.},\n bibtype = {article},\n author = {Loughlin, F.E. and Mansfield, R.E. and Vaz, P.M. and McGrath, A.P. and Setiyaputra, S. and Gamsjaeger, R. and Chen, E.S. and Morris, B.J. and Guss, J.M. and Mackay, J.P.},\n doi = {10.1073/pnas.0802466106},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {14}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The alternative splicing of mRNA is a critical process in higher eukaryotes that generates substantial proteomic diversity. Many of the proteins that are essential to this process contain arginine/serine-rich (RS) domains. ZRANB2 is a widely-expressed and highly-conserved RS-domain protein that can regulate alternative splicing but lacks canonical RNA-binding domains. Instead, it contains 2 RanBP2-type zinc finger (ZnF) domains. We demonstrate that these ZnFs recognize ssRNA with high affinity and specificity. Each ZnF binds to a single AGGUAA motif and the 2 domains combine to recognize AGGUAA (Nx) AGGUAA double sites, suggesting that ZRANB2 regulates alternative splicing via a direct interaction with pre-mRNA at sites that resemble the consensus 5' splice site. We show using X-ray crystallography that recognition of an AGGUAA motif by a single ZnF is dominated by side-chain hydrogen bonds to the bases and formation of a guanine-tryptophan-guanine ''ladder.'' A number of other human proteins that function in RNA processing also contain RanBP2 ZnFs in which the RNA-binding residues of ZRANB2 are conserved. The ZnFs of ZRANB2 therefore define another class of RNA-binding domain, advancing our understanding of RNA recognition and emphasizing the versatility of domains in molecular recognition.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"matthews-bhati-lehtomaki-mansfield-cubeddu-mackay-ittakestwototangothestructureandfunctionoflimringphdandmynddomains-2009\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  It takes two to tango: The structure and function of LIM, RING, PHD and MYND domains.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Matthews,  J.; Bhati,  M.; Lehtomaki,  E.; Mansfield,  R.; Cubeddu,  L.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Current Pharmaceutical Design</i>, 15(31): 3681-3696.  2009.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.2174/138161209789271861\"\n     onclick=\"log_download('matthews-bhati-lehtomaki-mansfield-cubeddu-mackay-ittakestwototangothestructureandfunctionoflimringphdandmynddomains-2009', 'http://doi.org/10.2174/138161209789271861', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/matthews-bhati-lehtomaki-mansfield-cubeddu-mackay-ittakestwototangothestructureandfunctionoflimringphdandmynddomains-2009\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('matthews-bhati-lehtomaki-mansfield-cubeddu-mackay-ittakestwototangothestructureandfunctionoflimringphdandmynddomains-2009')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {It takes two to tango: The structure and function of LIM, RING, PHD and MYND domains},\n type = {article},\n year = {2009},\n pages = {3681-3696},\n volume = {15},\n id = {12e1371d-f8f7-3ef4-8a33-189b9e2b23cc},\n created = {2023-01-10T01:45:32.880Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:32.880Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {LIM (Lin-11, Isl-1, Mec-3), RING (Really interesting new gene), PHD (Plant homology domain) and MYND (myeloid, Nervy, DEAF-1) domains are all zinc-binding domains that ligate two zinc ions. Unlike the better known classical zinc fingers, these domains do not bind DNA, but instead mediate interactions with other proteins. LIM-domain containing proteins have diverse functions as regulators of gene expression, cell adhesion and motility and signal transduction. RING finger proteins are generally associated with ubiquitination; the presence of such a domain is the defining feature of a class of E3 ubiquitin protein ligases. PHD proteins have been associated with SUMOylation but most recently have emerged as a chromatin recognition motif that reads the methylation state of histones. The function of the MYND domain is less clear, but MYND domains are also found in proteins that have ubiquitin ligase and/or histone methyltransferase activity. Here we review the structure-function relationships for these domains and discuss strategies to modulate their activity. © 2009 Bentham Science Publishers Ltd.},\n bibtype = {article},\n author = {Matthews, J.M. and Bhati, M. and Lehtomaki, E. and Mansfield, R.E. and Cubeddu, L. and Mackay, J.P.},\n doi = {10.2174/138161209789271861},\n journal = {Current Pharmaceutical Design},\n number = {31}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  LIM (Lin-11, Isl-1, Mec-3), RING (Really interesting new gene), PHD (Plant homology domain) and MYND (myeloid, Nervy, DEAF-1) domains are all zinc-binding domains that ligate two zinc ions. Unlike the better known classical zinc fingers, these domains do not bind DNA, but instead mediate interactions with other proteins. LIM-domain containing proteins have diverse functions as regulators of gene expression, cell adhesion and motility and signal transduction. RING finger proteins are generally associated with ubiquitination; the presence of such a domain is the defining feature of a class of E3 ubiquitin protein ligases. PHD proteins have been associated with SUMOylation but most recently have emerged as a chromatin recognition motif that reads the methylation state of histones. The function of the MYND domain is less clear, but MYND domains are also found in proteins that have ubiquitin ligase and/or histone methyltransferase activity. Here we review the structure-function relationships for these domains and discuss strategies to modulate their activity. © 2009 Bentham Science Publishers Ltd.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2008\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2008')\"\n      onkeypress=\"toggleGroup('group_2008')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2008</span>\n      <span class=\"bibbase_group_count\">\n        \n        (12)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://scripts.iucr.org/cgi-bin/paper?S1744309108036993\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008', 'http://scripts.iucr.org/cgi-bin/paper?S1744309108036993', event);\">\n    Crystallization of a ZRANB2–RNA complex.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Loughlin,  F., E.; Lee,  M.; Guss,  J., M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Acta Crystallographica Section F Structural Biology and Crystallization Communications</i>, 64(12): 1175-1177. 12 2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://scripts.iucr.org/cgi-bin/paper?S1744309108036993\"\n     onclick=\"log_download('loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008', 'http://scripts.iucr.org/cgi-bin/paper?S1744309108036993', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Crystallization of a ZRANB2–RNA complex [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1107/S1744309108036993\"\n     onclick=\"log_download('loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008', 'http://doi.org/10.1107/S1744309108036993', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Crystallization of a ZRANB2–RNA complex},\n type = {article},\n year = {2008},\n keywords = {RNA-binding proteins,RanBP2-type zinc fingers,Splicing factors},\n pages = {1175-1177},\n volume = {64},\n websites = {http://scripts.iucr.org/cgi-bin/paper?S1744309108036993},\n month = {12},\n day = {1},\n id = {7587f247-d4b3-314f-84e0-30baa59ddf19},\n created = {2020-12-17T05:29:52.864Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:52.864Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Loughlin2008},\n source_type = {ARTICLE},\n notes = {cited By 1},\n private_publication = {false},\n abstract = {ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.},\n bibtype = {article},\n author = {Loughlin, Fionna E. and Lee, Mihwa and Guss, J. Mitchell and Mackay, Joel P.},\n doi = {10.1107/S1744309108036993},\n journal = {Acta Crystallographica Section F Structural Biology and Crystallization Communications},\n number = {12}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"chu-gamsjaeger-mansfield-mackay-nmrspectroscopyasatoolfortherapidassessmentoftheconformationofgstfusionproteins-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  NMR spectroscopy as a tool for the rapid assessment of the conformation of GST-fusion proteins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Chu,  K.; Gamsjaeger,  R.; Mansfield,  R.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 17(9): 1630-1635.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1110/ps.034983.108\"\n     onclick=\"log_download('chu-gamsjaeger-mansfield-mackay-nmrspectroscopyasatoolfortherapidassessmentoftheconformationofgstfusionproteins-2008', 'http://doi.org/10.1110/ps.034983.108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/chu-gamsjaeger-mansfield-mackay-nmrspectroscopyasatoolfortherapidassessmentoftheconformationofgstfusionproteins-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('chu-gamsjaeger-mansfield-mackay-nmrspectroscopyasatoolfortherapidassessmentoftheconformationofgstfusionproteins-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {NMR spectroscopy as a tool for the rapid assessment of the conformation of GST-fusion proteins},\n type = {article},\n year = {2008},\n pages = {1630-1635},\n volume = {17},\n id = {2555f698-f625-3490-b864-b58850f3abab},\n created = {2023-01-10T01:45:33.906Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:33.906Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Glutathione-S-transferase (GST)-fusion proteins are used extensively for structural, biochemical, and functional analyses. Although the conformation of the target protein is of critical importance, confirmation of the folded state of the target is often not undertaken or is cumbersome because of the requirement to first remove the GST tag. Here, we demonstrate that it is possible to record conventional 15N-HSQC NMR spectra of small GST-fusion proteins and that the observed signals arise almost exclusively from the target protein. This approach constitutes a rapid and straightforward means of assessing the conformation of a GST-fusion protein without having to cleave the GST and should prove valuable, both to biochemists seeking to check the conformation of their proteins prior to functional studies and to structural biologists screening protein constructs for suitability as targets for structural studies. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 The Protein Society.},\n bibtype = {article},\n author = {Chu, K.L. and Gamsjaeger, R. and Mansfield, R.E. and Mackay, J.P.},\n doi = {10.1110/ps.034983.108},\n journal = {Protein Science},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Glutathione-S-transferase (GST)-fusion proteins are used extensively for structural, biochemical, and functional analyses. Although the conformation of the target protein is of critical importance, confirmation of the folded state of the target is often not undertaken or is cumbersome because of the requirement to first remove the GST tag. Here, we demonstrate that it is possible to record conventional 15N-HSQC NMR spectra of small GST-fusion proteins and that the observed signals arise almost exclusively from the target protein. This approach constitutes a rapid and straightforward means of assessing the conformation of a GST-fusion protein without having to cleave the GST and should prove valuable, both to biochemists seeking to check the conformation of their proteins prior to functional studies and to structural biologists screening protein constructs for suitability as targets for structural studies. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 The Protein Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"sunde-kwan-templeton-beever-mackay-structuralanalysisofhydrophobins-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural analysis of hydrophobins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Sunde,  M.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Templeton,  M.; Beever,  R.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Micron</i>, 39(7): 773-784.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.micron.2007.08.003\"\n     onclick=\"log_download('sunde-kwan-templeton-beever-mackay-structuralanalysisofhydrophobins-2008', 'http://doi.org/10.1016/j.micron.2007.08.003', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/sunde-kwan-templeton-beever-mackay-structuralanalysisofhydrophobins-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('sunde-kwan-templeton-beever-mackay-structuralanalysisofhydrophobins-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural analysis of hydrophobins},\n type = {article},\n year = {2008},\n pages = {773-784},\n volume = {39},\n id = {d6614c6a-3917-3fb1-bfca-b4688d016cf4},\n created = {2023-01-10T01:45:34.809Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:34.809Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Hydrophobins are a remarkable class of small cysteine-rich proteins found exclusively in fungi. They self-assemble to form robust polymeric monolayers that are highly amphipathic and play numerous roles in fungal biology, such as in the formation and dispersal of aerial spores and in pathogenic and mutualistic interactions. The polymeric form can be reversibly disassembled and is able to reverse the wettability of a surface, leading to many proposals for nanotechnological applications over recent years. The surprising properties of hydrophobins and their potential for commercialization have led to substantial efforts to delineate their morphology and molecular structure. In this review, we summarize the progress that has been made using a variety of spectroscopic and microscopic approaches towards understanding the molecular mechanisms underlying hydrophobin structure. © 2007 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Sunde, M. and Kwan, A.H.Y. and Templeton, M.D. and Beever, R.E. and Mackay, J.P.},\n doi = {10.1016/j.micron.2007.08.003},\n journal = {Micron},\n number = {7}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Hydrophobins are a remarkable class of small cysteine-rich proteins found exclusively in fungi. They self-assemble to form robust polymeric monolayers that are highly amphipathic and play numerous roles in fungal biology, such as in the formation and dispersal of aerial spores and in pathogenic and mutualistic interactions. The polymeric form can be reversibly disassembled and is able to reverse the wettability of a surface, leading to many proposals for nanotechnological applications over recent years. The surprising properties of hydrophobins and their potential for commercialization have led to substantial efforts to delineate their morphology and molecular structure. In this review, we summarize the progress that has been made using a variety of spectroscopic and microscopic approaches towards understanding the molecular mechanisms underlying hydrophobin structure. © 2007 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kwan-macindoe-vukain-morris-kass-gupte-mark-templeton-etal-thecys3cys4loopofthehydrophobineasisnotrequiredforrodletformationandsurfaceactivity-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The Cys3-Cys4 Loop of the Hydrophobin EAS Is Not Required for Rodlet Formation and Surface Activity.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kwan,  A.; Macindoe,  I.; Vukašin,  P.; Morris,  V.; Kass,  I.; Gupte,  R.; Mark,  A.; Templeton,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Sunde,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 382(3): 708-720.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2008.07.034\"\n     onclick=\"log_download('kwan-macindoe-vukain-morris-kass-gupte-mark-templeton-etal-thecys3cys4loopofthehydrophobineasisnotrequiredforrodletformationandsurfaceactivity-2008', 'http://doi.org/10.1016/j.jmb.2008.07.034', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kwan-macindoe-vukain-morris-kass-gupte-mark-templeton-etal-thecys3cys4loopofthehydrophobineasisnotrequiredforrodletformationandsurfaceactivity-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kwan-macindoe-vukain-morris-kass-gupte-mark-templeton-etal-thecys3cys4loopofthehydrophobineasisnotrequiredforrodletformationandsurfaceactivity-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The Cys3-Cys4 Loop of the Hydrophobin EAS Is Not Required for Rodlet Formation and Surface Activity},\n type = {article},\n year = {2008},\n pages = {708-720},\n volume = {382},\n id = {209bc497-4431-3395-a923-3ba1135b8fac},\n created = {2023-01-10T01:45:35.837Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:35.837Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Class I hydrophobins are fungal proteins that self-assemble into robust amphipathic rodlet monolayers on the surface of aerial structures such as spores and fruiting bodies. These layers share many structural characteristics with amyloid fibrils and belong to the growing family of functional amyloid-like materials produced by microorganisms. Although the three-dimensional structure of the soluble monomeric form of a class I hydrophobin has been determined, little is known about the molecular structure of the rodlets or their assembly mechanism. Several models have been proposed, some of which suggest that the Cys3-Cys4 loop has a critical role in the initiation of assembly or in the polymeric structure. In order to provide insight into the relationship between hydrophobin sequence and rodlet assembly, we investigated the role of the Cys3-Cys4 loop in EAS, a class I hydrophobin from Neurospora crassa. Remarkably, deletion of up to 15 residues from this 25-residue loop does not impair rodlet formation or reduce the surface activity of the protein, and the physicochemical properties of rodlets formed by this mutant are indistinguishable from those of its full-length counterpart. In addition, the core structure of the truncation mutant is essentially unchanged. Molecular dynamics simulations carried out on the full-length protein and this truncation mutant binding to an air-water interface show that, although it is hydrophobic, the loop does not play a role in positioning the protein at the surface. These results demonstrate that the Cys3-Cys4 loop does not have an integral role in the formation or structure of the rodlets and that the major determinant of the unique properties of these proteins is the amphipathic core structure, which is likely to be preserved in all hydrophobins despite the high degree of sequence variation across the family. © 2008 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Kwan, A.H. and Macindoe, I. and Vukašin, P.V. and Morris, V.K. and Kass, I. and Gupte, R. and Mark, A.E. and Templeton, M.D. and Mackay, J.P. and Sunde, M.},\n doi = {10.1016/j.jmb.2008.07.034},\n journal = {Journal of Molecular Biology},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Class I hydrophobins are fungal proteins that self-assemble into robust amphipathic rodlet monolayers on the surface of aerial structures such as spores and fruiting bodies. These layers share many structural characteristics with amyloid fibrils and belong to the growing family of functional amyloid-like materials produced by microorganisms. Although the three-dimensional structure of the soluble monomeric form of a class I hydrophobin has been determined, little is known about the molecular structure of the rodlets or their assembly mechanism. Several models have been proposed, some of which suggest that the Cys3-Cys4 loop has a critical role in the initiation of assembly or in the polymeric structure. In order to provide insight into the relationship between hydrophobin sequence and rodlet assembly, we investigated the role of the Cys3-Cys4 loop in EAS, a class I hydrophobin from Neurospora crassa. Remarkably, deletion of up to 15 residues from this 25-residue loop does not impair rodlet formation or reduce the surface activity of the protein, and the physicochemical properties of rodlets formed by this mutant are indistinguishable from those of its full-length counterpart. In addition, the core structure of the truncation mutant is essentially unchanged. Molecular dynamics simulations carried out on the full-length protein and this truncation mutant binding to an air-water interface show that, although it is hydrophobic, the loop does not play a role in positioning the protein at the surface. These results demonstrate that the Cys3-Cys4 loop does not have an integral role in the formation or structure of the rodlets and that the major determinant of the unique properties of these proteins is the amphipathic core structure, which is likely to be preserved in all hydrophobins despite the high degree of sequence variation across the family. © 2008 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-sunde-lowry-crossley-matthews-responsetochatraryamontrietalproteininteractionstobelieveornottobelieve-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Response to Chatr-aryamontri et al.: Protein interactions: to believe or not to believe?.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Sunde,  M.; Lowry,  J.; Crossley,  M.;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Trends in Biochemical Sciences</i>, 33(6): 242-243.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.tibs.2008.04.003\"\n     onclick=\"log_download('mackay-sunde-lowry-crossley-matthews-responsetochatraryamontrietalproteininteractionstobelieveornottobelieve-2008', 'http://doi.org/10.1016/j.tibs.2008.04.003', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-sunde-lowry-crossley-matthews-responsetochatraryamontrietalproteininteractionstobelieveornottobelieve-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-sunde-lowry-crossley-matthews-responsetochatraryamontrietalproteininteractionstobelieveornottobelieve-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Response to Chatr-aryamontri et al.: Protein interactions: to believe or not to believe?},\n type = {article},\n year = {2008},\n pages = {242-243},\n volume = {33},\n id = {a93b20e9-5f78-368e-9bc7-126a1f8e9c4e},\n created = {2023-01-10T01:45:36.806Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:36.806Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Mackay, J.P. and Sunde, M. and Lowry, J.A. and Crossley, M. and Matthews, J.M.},\n doi = {10.1016/j.tibs.2008.04.003},\n journal = {Trends in Biochemical Sciences},\n number = {6}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"griffin-dobson-pearce-antonio-whitten-liew-mackay-trewhella-etal-evolutionofquaternarystructureinahomotetramericenzyme-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Evolution of Quaternary Structure in a Homotetrameric Enzyme.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Griffin,  M.; Dobson,  R.; Pearce,  F.; Antonio,  L.; Whitten,  A.; Liew,  C.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Trewhella,  J.; Jameson,  G.; Perugini,  M.; Perugini,  M.;  and Gerrard,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 380(4): 691-703.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2008.05.038\"\n     onclick=\"log_download('griffin-dobson-pearce-antonio-whitten-liew-mackay-trewhella-etal-evolutionofquaternarystructureinahomotetramericenzyme-2008', 'http://doi.org/10.1016/j.jmb.2008.05.038', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/griffin-dobson-pearce-antonio-whitten-liew-mackay-trewhella-etal-evolutionofquaternarystructureinahomotetramericenzyme-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('griffin-dobson-pearce-antonio-whitten-liew-mackay-trewhella-etal-evolutionofquaternarystructureinahomotetramericenzyme-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Evolution of Quaternary Structure in a Homotetrameric Enzyme},\n type = {article},\n year = {2008},\n pages = {691-703},\n volume = {380},\n id = {63b6023f-2683-3fd8-a96d-6ee13c0b2a51},\n created = {2023-01-10T01:45:37.717Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:37.717Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Dihydrodipicolinate synthase (DHDPS) is an essential enzyme in (S)-lysine biosynthesis and an important antibiotic target. All X-ray crystal structures solved to date reveal a homotetrameric enzyme. In order to explore the role of this quaternary structure, dimeric variants of Escherichia coli DHDPS were engineered and their properties were compared to those of the wild-type tetrameric form. X-ray crystallography reveals that the active site is not disturbed when the quaternary structure is disrupted. However, the activity of the dimeric enzymes in solution is substantially reduced, and a tetrahedral adduct of a substrate analogue is observed to be trapped at the active site in the crystal form. Remarkably, heating the dimeric enzymes increases activity. We propose that the homotetrameric structure of DHDPS reduces dynamic fluctuations present in the dimeric forms and increases specificity for the first substrate, pyruvate. By restricting motion in a key catalytic motif, a competing, non-productive reaction with a substrate analogue is avoided. Small-angle X-ray scattering and mutagenesis data, together with a B-factor analysis of the crystal structures, support this hypothesis and lead to the suggestion that in at least some cases, the evolution of quaternary enzyme structures might serve to optimise the dynamic properties of the protein subunits. © 2008 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Griffin, M.D.W. and Dobson, R.C.J. and Pearce, F.G. and Antonio, L. and Whitten, A.E. and Liew, C.K. and Mackay, J.P. and Trewhella, J. and Jameson, G.B. and Perugini, M.A. and Perugini, M.A. and Gerrard, J.A.},\n doi = {10.1016/j.jmb.2008.05.038},\n journal = {Journal of Molecular Biology},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Dihydrodipicolinate synthase (DHDPS) is an essential enzyme in (S)-lysine biosynthesis and an important antibiotic target. All X-ray crystal structures solved to date reveal a homotetrameric enzyme. In order to explore the role of this quaternary structure, dimeric variants of Escherichia coli DHDPS were engineered and their properties were compared to those of the wild-type tetrameric form. X-ray crystallography reveals that the active site is not disturbed when the quaternary structure is disrupted. However, the activity of the dimeric enzymes in solution is substantially reduced, and a tetrahedral adduct of a substrate analogue is observed to be trapped at the active site in the crystal form. Remarkably, heating the dimeric enzymes increases activity. We propose that the homotetrameric structure of DHDPS reduces dynamic fluctuations present in the dimeric forms and increases specificity for the first substrate, pyruvate. By restricting motion in a key catalytic motif, a competing, non-productive reaction with a substrate analogue is avoided. Small-angle X-ray scattering and mutagenesis data, together with a B-factor analysis of the crystal structures, support this hypothesis and lead to the suggestion that in at least some cases, the evolution of quaternary enzyme structures might serve to optimise the dynamic properties of the protein subunits. © 2008 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"bhati-lee-nancarrow-lee-craig-bach-guss-mackay-etal-implementingthelimcodethestructuralbasisforcelltypespecificassemblyoflimhomeodomaincomplexes-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Implementing the LIM code: The structural basis for cell type-specific assembly of LIM-homeodomain complexes.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Bhati,  M.; Lee,  C.; Nancarrow,  A.; Lee,  M.; Craig,  V.; Bach,  I.; Guss,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>EMBO Journal</i>, 27(14): 2018-2029.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/emboj.2008.123\"\n     onclick=\"log_download('bhati-lee-nancarrow-lee-craig-bach-guss-mackay-etal-implementingthelimcodethestructuralbasisforcelltypespecificassemblyoflimhomeodomaincomplexes-2008', 'http://doi.org/10.1038/emboj.2008.123', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/bhati-lee-nancarrow-lee-craig-bach-guss-mackay-etal-implementingthelimcodethestructuralbasisforcelltypespecificassemblyoflimhomeodomaincomplexes-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('bhati-lee-nancarrow-lee-craig-bach-guss-mackay-etal-implementingthelimcodethestructuralbasisforcelltypespecificassemblyoflimhomeodomaincomplexes-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Implementing the LIM code: The structural basis for cell type-specific assembly of LIM-homeodomain complexes},\n type = {article},\n year = {2008},\n pages = {2018-2029},\n volume = {27},\n id = {aab1c421-98b0-388e-a789-d78011488b4f},\n created = {2023-01-10T01:45:38.632Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:38.632Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {LIM-homeodomain (LIM-HD) transcription factors form a combinatorial &#x27;LIM code&#x27; that contributes to the specification of cell types. In the ventral spinal cord, the binary LIM homeobox protein 3 (Lhx3)/LIM domain-binding protein 1 (Ldb1) complex specifies the formation of V2 interneurons. The additional expression of islet-1 (Isl1) in adjacent cells instead specifies the formation of motor neurons through assembly of a ternary complex in which Isl1 contacts both Lhx3 and Ldb1, displacing Lhx3 as the binding partner of Ldb1. However, little is known about how this molecular switch occurs. Here, we have identified the 30-residue Lhx3-binding domain on Isl1 (Isl1LBD). Although the LIM interaction domain of Ldb1 (Ldb1LID) and Isl1LBD share low levels of sequence homology, X-ray and NMR structures reveal that they bind Lhx3 in an identical manner, that is, Isl1LBD mimics Ldb1 LID. These data provide a structural basis for the formation of cell type-specific protein-protein interactions in which unstructured linear motifs with diverse sequences compete to bind protein partners. The resulting alternate protein complexes can target different genes to regulate key biological events. ©2008 European Molecular Biology Organization.},\n bibtype = {article},\n author = {Bhati, M. and Lee, C. and Nancarrow, A.L. and Lee, M. and Craig, V.J. and Bach, I. and Guss, J.M. and Mackay, J.P. and Matthews, J.M.},\n doi = {10.1038/emboj.2008.123},\n journal = {EMBO Journal},\n number = {14}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  LIM-homeodomain (LIM-HD) transcription factors form a combinatorial 'LIM code' that contributes to the specification of cell types. In the ventral spinal cord, the binary LIM homeobox protein 3 (Lhx3)/LIM domain-binding protein 1 (Ldb1) complex specifies the formation of V2 interneurons. The additional expression of islet-1 (Isl1) in adjacent cells instead specifies the formation of motor neurons through assembly of a ternary complex in which Isl1 contacts both Lhx3 and Ldb1, displacing Lhx3 as the binding partner of Ldb1. However, little is known about how this molecular switch occurs. Here, we have identified the 30-residue Lhx3-binding domain on Isl1 (Isl1LBD). Although the LIM interaction domain of Ldb1 (Ldb1LID) and Isl1LBD share low levels of sequence homology, X-ray and NMR structures reveal that they bind Lhx3 in an identical manner, that is, Isl1LBD mimics Ldb1 LID. These data provide a structural basis for the formation of cell type-specific protein-protein interactions in which unstructured linear motifs with diverse sequences compete to bind protein partners. The resulting alternate protein complexes can target different genes to regulate key biological events. ©2008 European Molecular Biology Organization.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gamsjaeger-swanton-kobus-lehtomaki-lowry-kwan-matthews-mackay-structuralandbiophysicalanalysisofthednabindingpropertiesofmyelintranscriptionfactor1-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural and biophysical analysis of the DNA binding properties of myelin transcription factor 1.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gamsjaeger,  R.; Swanton,  M.; Kobus,  F.; Lehtomaki,  E.; Lowry,  J.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Matthews,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 283(8): 5158-5167.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M703772200\"\n     onclick=\"log_download('gamsjaeger-swanton-kobus-lehtomaki-lowry-kwan-matthews-mackay-structuralandbiophysicalanalysisofthednabindingpropertiesofmyelintranscriptionfactor1-2008', 'http://doi.org/10.1074/jbc.M703772200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gamsjaeger-swanton-kobus-lehtomaki-lowry-kwan-matthews-mackay-structuralandbiophysicalanalysisofthednabindingpropertiesofmyelintranscriptionfactor1-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gamsjaeger-swanton-kobus-lehtomaki-lowry-kwan-matthews-mackay-structuralandbiophysicalanalysisofthednabindingpropertiesofmyelintranscriptionfactor1-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural and biophysical analysis of the DNA binding properties of myelin transcription factor 1},\n type = {article},\n year = {2008},\n pages = {5158-5167},\n volume = {283},\n id = {915fd038-e9b1-36a4-8be4-dcecb9f56909},\n created = {2023-01-10T01:45:39.551Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:39.551Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Zinc binding domains, or zinc fingers (ZnFs), form one of the most numerous and most diverse superclasses of protein structural motifs in eukaryotes. Although our understanding of the functions of several classes of these domains is relatively well developed, we know much less about the molecular mechanisms of action of many others. Myelin transcription factor 1 (MyT1) type ZnFs are found in organisms as diverse as nematodes and mammals and are found in a range of sequence contexts. MyT1, one of the early transcription factors expressed in the developing central nervous system, contains seven MyT1 ZnFs that are very highly conserved both within the protein and between species. We have used a range of biophysical techniques, including NMR spectroscopy and data-driven macromolecular docking, to investigate the structural basis for the interaction between MyT1 ZnFs and DNA. Our data indicate that MyT1 ZnFs recognize the major groove of DNA in a way that appears to differ from other known zinc binding domains. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Gamsjaeger, R. and Swanton, M.K. and Kobus, F.J. and Lehtomaki, E. and Lowry, J.A. and Kwan, A.H. and Matthews, J.M. and Mackay, J.P.},\n doi = {10.1074/jbc.M703772200},\n journal = {Journal of Biological Chemistry},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Zinc binding domains, or zinc fingers (ZnFs), form one of the most numerous and most diverse superclasses of protein structural motifs in eukaryotes. Although our understanding of the functions of several classes of these domains is relatively well developed, we know much less about the molecular mechanisms of action of many others. Myelin transcription factor 1 (MyT1) type ZnFs are found in organisms as diverse as nematodes and mammals and are found in a range of sequence contexts. MyT1, one of the early transcription factors expressed in the developing central nervous system, contains seven MyT1 ZnFs that are very highly conserved both within the protein and between species. We have used a range of biophysical techniques, including NMR spectroscopy and data-driven macromolecular docking, to investigate the structural basis for the interaction between MyT1 ZnFs and DNA. Our data indicate that MyT1 ZnFs recognize the major groove of DNA in a way that appears to differ from other known zinc binding domains. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"langley-shojaei-chan-hiu-mackay-traut-guss-christopherson-structureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparum-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structure and inhibition of orotidine 5′-monophosphate decarboxylase from Plasmodium falciparum.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Langley,  D.; Shojaei,  M.; Chan,  C.; Hiu,  C.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Traut,  T.; Guss,  J.;  and Christopherson,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 47(12): 3842-3854.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi702390k\"\n     onclick=\"log_download('langley-shojaei-chan-hiu-mackay-traut-guss-christopherson-structureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparum-2008', 'http://doi.org/10.1021/bi702390k', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/langley-shojaei-chan-hiu-mackay-traut-guss-christopherson-structureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparum-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('langley-shojaei-chan-hiu-mackay-traut-guss-christopherson-structureandinhibitionoforotidine5monophosphatedecarboxylasefromplasmodiumfalciparum-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structure and inhibition of orotidine 5′-monophosphate decarboxylase from Plasmodium falciparum},\n type = {article},\n year = {2008},\n pages = {3842-3854},\n volume = {47},\n id = {a141dd55-ab8c-3d60-ae66-5bb20bb77e87},\n created = {2023-01-10T01:45:40.465Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:40.465Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Orotidine 5′-monophosphate (OMP) decarboxylase from Plasmodium falciparum (PfODCase, EC 4.1.1.23) has been overexpressed, purified, subjected to kinetic and biochemical analysis, and crystallized. The native enzyme is a homodimer with a subunit molecular mass of 38 kDa. The saturation curve for OMP as a substrate conformed to Michaelis-Menten kinetics with Km = 350 ± 60 nM and Vmax = 2.70 ± 0.10 μmol/min/mg protein. Inhibition patterns for nucleoside 5′-monophosphate analogues were linear competitive with respect to OMP with a decreasing potency of inhibition of PfODCase in the order: pyrazofurin 5′-monophosphate (Ki = 3.6 ± 0.7 nM) &gt; xanthosine 5′-monophosphate (XMP, Ki = 4.4 ± 0.7 nM) &gt; 6-azauridine 5′-monophosphate (AzaUMP, K i = 12 ±3 nM) &gt; allopurinol-3-riboside 5′- monophosphate (Ki = 240 ± 20 nM). XMP is an ∼150-fold more potent inhibitor of PfODCase compared with the human enzyme. The structure of PfODCase was solved in the absence of ligand and displays a classic TIM-barrel fold characteristic of the enzyme. Both the phosphate-binding loop and the βα5-loop have conformational flexibility, which may be associated with substrate capture and product release along the reaction pathway. © 2008 American Chemical Society.},\n bibtype = {article},\n author = {Langley, D.B. and Shojaei, M. and Chan, C. and Hiu, C.L. and Mackay, J.P. and Traut, T.W. and Guss, J.M. and Christopherson, R.I.},\n doi = {10.1021/bi702390k},\n journal = {Biochemistry},\n number = {12}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Orotidine 5′-monophosphate (OMP) decarboxylase from Plasmodium falciparum (PfODCase, EC 4.1.1.23) has been overexpressed, purified, subjected to kinetic and biochemical analysis, and crystallized. The native enzyme is a homodimer with a subunit molecular mass of 38 kDa. The saturation curve for OMP as a substrate conformed to Michaelis-Menten kinetics with Km = 350 ± 60 nM and Vmax = 2.70 ± 0.10 μmol/min/mg protein. Inhibition patterns for nucleoside 5′-monophosphate analogues were linear competitive with respect to OMP with a decreasing potency of inhibition of PfODCase in the order: pyrazofurin 5′-monophosphate (Ki = 3.6 ± 0.7 nM) > xanthosine 5′-monophosphate (XMP, Ki = 4.4 ± 0.7 nM) > 6-azauridine 5′-monophosphate (AzaUMP, K i = 12 ±3 nM) > allopurinol-3-riboside 5′- monophosphate (Ki = 240 ± 20 nM). XMP is an ∼150-fold more potent inhibitor of PfODCase compared with the human enzyme. The structure of PfODCase was solved in the absence of ligand and displays a classic TIM-barrel fold characteristic of the enzyme. Both the phosphate-binding loop and the βα5-loop have conformational flexibility, which may be associated with substrate capture and product release along the reaction pathway. © 2008 American Chemical Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Crystallization of a ZRANB2-RNA complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Loughlin,  F.; Lee,  M.; Guss,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Acta Crystallographica Section F: Structural Biology and Crystallization Communications</i>, 64(12): 1175-1177.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1107/S1744309108036993\"\n     onclick=\"log_download('loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008', 'http://doi.org/10.1107/S1744309108036993', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('loughlin-lee-guss-mackay-crystallizationofazranb2rnacomplex-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Crystallization of a ZRANB2-RNA complex},\n type = {article},\n year = {2008},\n pages = {1175-1177},\n volume = {64},\n id = {5c489013-8140-39aa-85b6-66fd1e85cec0},\n created = {2023-01-10T01:45:41.371Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:41.371Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.},\n bibtype = {article},\n author = {Loughlin, F.E. and Lee, M. and Guss, J.M. and Mackay, J.P.},\n doi = {10.1107/S1744309108036993},\n journal = {Acta Crystallographica Section F: Structural Biology and Crystallization Communications},\n number = {12}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  ZRANB2 is a zinc-finger protein that has been shown to influence alternative splice-site selection. The protein comprises a C-terminal arginine/serine-rich domain that interacts with spliceosomal proteins and two N-terminal RanBP2-type zinc fingers that have been implicated in RNA recognition. The second zinc finger bound to a six-nucleotide single-stranded RNA target sequence crystallized in the hexagonal space group P6522 or P6122, with unit-cell parameters a = 54.52, b = 54.52, c = 48.07 Å; the crystal contains one monomeric complex per asymmetric unit. This crystal form has a solvent content of 39% and diffracted to 1.4 Å resolution using synchrotron radiation. © International Union of Crystallography 2008.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"matthews-loughlin-mackay-designedmetalbindingsitesinbiomolecularandbioinorganicinteractions-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Designed metal-binding sites in biomolecular and bioinorganic interactions.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Matthews,  J.; Loughlin,  F.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Current Opinion in Structural Biology</i>, 18(4): 484-490.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.sbi.2008.04.009\"\n     onclick=\"log_download('matthews-loughlin-mackay-designedmetalbindingsitesinbiomolecularandbioinorganicinteractions-2008', 'http://doi.org/10.1016/j.sbi.2008.04.009', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/matthews-loughlin-mackay-designedmetalbindingsitesinbiomolecularandbioinorganicinteractions-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('matthews-loughlin-mackay-designedmetalbindingsitesinbiomolecularandbioinorganicinteractions-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Designed metal-binding sites in biomolecular and bioinorganic interactions},\n type = {article},\n year = {2008},\n pages = {484-490},\n volume = {18},\n id = {719ca520-9316-3d73-84b3-1e92163b1c32},\n created = {2023-01-10T01:45:42.283Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:42.283Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The design of metal-binding functionality in proteins is expanding into many different areas with a wide range of practical and research applications. Here we review several developing areas of metal-related protein design, including the use of metals to induce protein-protein interactions or facilitate the assembly of extended nanostructures; the design of metallopeptides that bind metal and other inorganic surfaces, an area with potential in diverse applications ranging from nanoelectronics and photonics to biotechnology and biomedicine; and, the creation of sensitive and selective metal sensors for use both in vivo and in vitro. © 2008 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Matthews, J.M. and Loughlin, F.E. and Mackay, J.P.},\n doi = {10.1016/j.sbi.2008.04.009},\n journal = {Current Opinion in Structural Biology},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The design of metal-binding functionality in proteins is expanding into many different areas with a wide range of practical and research applications. Here we review several developing areas of metal-related protein design, including the use of metals to induce protein-protein interactions or facilitate the assembly of extended nanostructures; the design of metallopeptides that bind metal and other inorganic surfaces, an area with potential in diverse applications ranging from nanoelectronics and photonics to biotechnology and biomedicine; and, the creation of sensitive and selective metal sensors for use both in vivo and in vitro. © 2008 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wilkinson-liew-mackay-salleh-withers-mcleod-escherichiacoliglucuronylsynthaseanengineeredenzymeforthesynthesisofglucuronides-2008\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Escherichia coli glucuronylsynthase: An engineered enzyme for the synthesis of β-glucuronides.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wilkinson,  S.; Liew,  C.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Salleh,  H.; Withers,  S.;  and McLeod,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Organic Letters</i>, 10(8): 1585-1588.  2008.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/ol8002767\"\n     onclick=\"log_download('wilkinson-liew-mackay-salleh-withers-mcleod-escherichiacoliglucuronylsynthaseanengineeredenzymeforthesynthesisofglucuronides-2008', 'http://doi.org/10.1021/ol8002767', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wilkinson-liew-mackay-salleh-withers-mcleod-escherichiacoliglucuronylsynthaseanengineeredenzymeforthesynthesisofglucuronides-2008\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wilkinson-liew-mackay-salleh-withers-mcleod-escherichiacoliglucuronylsynthaseanengineeredenzymeforthesynthesisofglucuronides-2008')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Escherichia coli glucuronylsynthase: An engineered enzyme for the synthesis of β-glucuronides},\n type = {article},\n year = {2008},\n pages = {1585-1588},\n volume = {10},\n id = {d014882a-497b-3d62-96a2-9f6d9e1e973d},\n created = {2023-01-10T01:45:43.190Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:43.190Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The glycosynthase derived from E. coli β-glucuronidase catalyzes the glucuronylation of a range of primary, secondary, and aryl alcohols with moderate to excellent yields. The procedure provides an efficient, stereoselective, and scalable single-step synthesis of β-glucuronides under mild conditions. © 2008 American Chemical Society.},\n bibtype = {article},\n author = {Wilkinson, S.M. and Liew, C.W. and Mackay, J.P. and Salleh, H.M. and Withers, S.G. and McLeod, M.D.},\n doi = {10.1021/ol8002767},\n journal = {Organic Letters},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The glycosynthase derived from E. coli β-glucuronidase catalyzes the glucuronylation of a range of primary, secondary, and aryl alcohols with moderate to excellent yields. The procedure provides an efficient, stereoselective, and scalable single-step synthesis of β-glucuronides under mild conditions. © 2008 American Chemical Society.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2007\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2007')\"\n      onkeypress=\"toggleGroup('group_2007')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2007</span>\n      <span class=\"bibbase_group_count\">\n        \n        (7)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"porter-matthews-mackay-pursglove-schmidberger-leedman-pero-krag-etal-grb7sh2domainstructureandinteractionswithacyclicpeptideinhibitorofcancercellmigrationandproliferation-2007\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Grb7 SH2 domain structure and interactions with a cyclic peptide inhibitor of cancer cell migration and proliferation.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Porter,  C.; Matthews,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Pursglove,  S.; Schmidberger,  J.; Leedman,  P.; Pero,  S.; Krag,  D.; Wilce,  M.;  and Wilce,  J.\n</span>\n\n  \n\n</span>\n\n  <i>BMC Structural Biology</i>, 7.  2007.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1186/1472-6807-7-58\"\n     onclick=\"log_download('porter-matthews-mackay-pursglove-schmidberger-leedman-pero-krag-etal-grb7sh2domainstructureandinteractionswithacyclicpeptideinhibitorofcancercellmigrationandproliferation-2007', 'http://doi.org/10.1186/1472-6807-7-58', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/porter-matthews-mackay-pursglove-schmidberger-leedman-pero-krag-etal-grb7sh2domainstructureandinteractionswithacyclicpeptideinhibitorofcancercellmigrationandproliferation-2007\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('porter-matthews-mackay-pursglove-schmidberger-leedman-pero-krag-etal-grb7sh2domainstructureandinteractionswithacyclicpeptideinhibitorofcancercellmigrationandproliferation-2007')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Grb7 SH2 domain structure and interactions with a cyclic peptide inhibitor of cancer cell migration and proliferation},\n type = {article},\n year = {2007},\n volume = {7},\n id = {4beae229-a038-3821-9cc9-5285dcb8776b},\n created = {2023-01-10T01:45:44.136Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:44.136Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background. Human growth factor receptor bound protein 7 (Grb7) is an adapter protein that mediates the coupling of tyrosine kinases with their downstream signaling pathways. Grb7 is frequently overexpressed in invasive and metastatic human cancers and is implicated in cancer progression via its interaction with the ErbB2 receptor and focal adhesion kinase (FAK) that play critical roles in cell proliferation and migration. It is thus a prime target for the development of novel anti-cancer therapies. Recently, an inhibitory peptide (G7-18NATE) has been developed which binds specifically to the Grb7 SH2 domain and is able to attenuate cancer cell proliferation and migration in various cancer cell lines. Results. As a first step towards understanding how Grb7 may be inhibited by G7-18NATE, we solved the crystal structure of the Grb7 SH2 domain to 2.1 Å resolution. We describe the details of the peptide binding site underlying target specificity, as well as the dimer interface of Grb 7 SH2. Dimer formation of Grb7 was determined to be in the μM range using analytical ultracentrifugation for both full-length Grb7 and the SH2 domain alone, suggesting the SH2 domain forms the basis of a physiological dimer. ITC measurements of the interaction of the G7-18NATE peptide with the Grb7 SH2 domain revealed that it binds with a binding affinity of Kd= ∼35.7 μM and NMR spectroscopy titration experiments revealed that peptide binding causes perturbations to both the ligand binding surface of the Grb7 SH2 domain as well as to the dimer interface, suggesting that dimerisation of Grb7 is impacted on by peptide binding. Conclusion. Together the data allow us to propose a model of the Grb7 SH2 domain/G7-18NATE interaction and to rationalize the basis for the observed binding specificity and affinity. We propose that the current study will assist with the development of second generation Grb7 SH2 domain inhibitors, potentially leading to novel inhibitors of cancer cell migration and invasion. © 2007 Porter et al; licensee BioMed Central Ltd.},\n bibtype = {article},\n author = {Porter, C.J. and Matthews, J.M. and Mackay, J.P. and Pursglove, S.E. and Schmidberger, J.W. and Leedman, P.J. and Pero, S.C. and Krag, D.N. and Wilce, M.C.J. and Wilce, J.A.},\n doi = {10.1186/1472-6807-7-58},\n journal = {BMC Structural Biology}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background. Human growth factor receptor bound protein 7 (Grb7) is an adapter protein that mediates the coupling of tyrosine kinases with their downstream signaling pathways. Grb7 is frequently overexpressed in invasive and metastatic human cancers and is implicated in cancer progression via its interaction with the ErbB2 receptor and focal adhesion kinase (FAK) that play critical roles in cell proliferation and migration. It is thus a prime target for the development of novel anti-cancer therapies. Recently, an inhibitory peptide (G7-18NATE) has been developed which binds specifically to the Grb7 SH2 domain and is able to attenuate cancer cell proliferation and migration in various cancer cell lines. Results. As a first step towards understanding how Grb7 may be inhibited by G7-18NATE, we solved the crystal structure of the Grb7 SH2 domain to 2.1 Å resolution. We describe the details of the peptide binding site underlying target specificity, as well as the dimer interface of Grb 7 SH2. Dimer formation of Grb7 was determined to be in the μM range using analytical ultracentrifugation for both full-length Grb7 and the SH2 domain alone, suggesting the SH2 domain forms the basis of a physiological dimer. ITC measurements of the interaction of the G7-18NATE peptide with the Grb7 SH2 domain revealed that it binds with a binding affinity of Kd= ∼35.7 μM and NMR spectroscopy titration experiments revealed that peptide binding causes perturbations to both the ligand binding surface of the Grb7 SH2 domain as well as to the dimer interface, suggesting that dimerisation of Grb7 is impacted on by peptide binding. Conclusion. Together the data allow us to propose a model of the Grb7 SH2 domain/G7-18NATE interaction and to rationalize the basis for the observed binding specificity and affinity. We propose that the current study will assist with the development of second generation Grb7 SH2 domain inhibitors, potentially leading to novel inhibitors of cancer cell migration and invasion. © 2007 Porter et al; licensee BioMed Central Ltd.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gamsjaeger-liew-loughlin-crossley-mackay-stickyfingerszincfingersasproteinrecognitionmotifs-2007\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Sticky fingers: zinc-fingers as protein-recognition motifs.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gamsjaeger,  R.; Liew,  C.; Loughlin,  F.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Trends in Biochemical Sciences</i>, 32(2): 63-70.  2007.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.tibs.2006.12.007\"\n     onclick=\"log_download('gamsjaeger-liew-loughlin-crossley-mackay-stickyfingerszincfingersasproteinrecognitionmotifs-2007', 'http://doi.org/10.1016/j.tibs.2006.12.007', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gamsjaeger-liew-loughlin-crossley-mackay-stickyfingerszincfingersasproteinrecognitionmotifs-2007\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gamsjaeger-liew-loughlin-crossley-mackay-stickyfingerszincfingersasproteinrecognitionmotifs-2007')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Sticky fingers: zinc-fingers as protein-recognition motifs},\n type = {article},\n year = {2007},\n pages = {63-70},\n volume = {32},\n id = {0bdbbb8b-8f2a-39d3-a9c6-8cef76dd27e8},\n created = {2023-01-10T01:45:45.060Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:45.060Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Zinc-fingers (ZnFs) are extremely abundant in higher eukaryotes. Once considered to function exclusively as sequence-specific DNA-binding motifs, ZnFs are now known to have additional activities such as the recognition of RNA and other proteins. Here we discuss recent advances in our understanding of ZnFs as specific modules for protein recognition. Structural studies of ZnF complexes reveal considerable diversity in terms of protein partners, binding modes and affinities, and highlight the often underestimated versatility of ZnF structure and function. An appreciation of the structural features of ZnF-protein interactions will contribute to our ability to engineer and to use ZnFs with tailored protein-binding properties. © 2006 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Gamsjaeger, R. and Liew, C.K. and Loughlin, F.E. and Crossley, M. and Mackay, J.P.},\n doi = {10.1016/j.tibs.2006.12.007},\n journal = {Trends in Biochemical Sciences},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Zinc-fingers (ZnFs) are extremely abundant in higher eukaryotes. Once considered to function exclusively as sequence-specific DNA-binding motifs, ZnFs are now known to have additional activities such as the recognition of RNA and other proteins. Here we discuss recent advances in our understanding of ZnFs as specific modules for protein recognition. Structural studies of ZnF complexes reveal considerable diversity in terms of protein partners, binding modes and affinities, and highlight the often underestimated versatility of ZnF structure and function. An appreciation of the structural features of ZnF-protein interactions will contribute to our ability to engineer and to use ZnFs with tailored protein-binding properties. © 2006 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"liew-crossley-mackay-nicholas-solutionstructureofthethapdomainfromcaenorhabditiseleganscterminalbindingproteinctbp-2007\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Solution Structure of the THAP Domain from Caenorhabditis elegans C-terminal Binding Protein (CtBP).\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Liew,  C.; Crossley,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Nicholas,  H.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 366(2): 382-390.  2007.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2006.11.058\"\n     onclick=\"log_download('liew-crossley-mackay-nicholas-solutionstructureofthethapdomainfromcaenorhabditiseleganscterminalbindingproteinctbp-2007', 'http://doi.org/10.1016/j.jmb.2006.11.058', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/liew-crossley-mackay-nicholas-solutionstructureofthethapdomainfromcaenorhabditiseleganscterminalbindingproteinctbp-2007\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('liew-crossley-mackay-nicholas-solutionstructureofthethapdomainfromcaenorhabditiseleganscterminalbindingproteinctbp-2007')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution Structure of the THAP Domain from Caenorhabditis elegans C-terminal Binding Protein (CtBP)},\n type = {article},\n year = {2007},\n pages = {382-390},\n volume = {366},\n id = {5e95220d-3a32-3190-836d-ac33617c7af2},\n created = {2023-01-10T01:45:45.962Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:45.962Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The THAP (Thanatos-associated protein) domain is a recently discovered zinc-binding domain found in proteins involved in transcriptional regulation, cell-cycle control, apoptosis and chromatin modification. It contains a single zinc atom ligated by cysteine and histidine residues within a Cys-X2-4-Cys-X35-53-Cys-X2-His consensus. We have determined the NMR solution structure of the THAP domain from Caenorhabditis elegans C-terminal binding protein (CtBP) and show that it adopts a fold containing a treble clef motif, bearing similarity to the zinc finger-associated domain (ZAD) from Drosophila Grauzone. The CtBP THAP domain contains a large, positively charged surface patch and we demonstrate that this domain can bind to double-stranded DNA in an electrophoretic mobility-shift assay. These data, together with existing reports, indicate that THAP domains might exhibit a functional diversity similar to that observed for classical and GATA-type zinc fingers. © 2006 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Liew, C.K. and Crossley, M. and Mackay, J.P. and Nicholas, H.R.},\n doi = {10.1016/j.jmb.2006.11.058},\n journal = {Journal of Molecular Biology},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The THAP (Thanatos-associated protein) domain is a recently discovered zinc-binding domain found in proteins involved in transcriptional regulation, cell-cycle control, apoptosis and chromatin modification. It contains a single zinc atom ligated by cysteine and histidine residues within a Cys-X2-4-Cys-X35-53-Cys-X2-His consensus. We have determined the NMR solution structure of the THAP domain from Caenorhabditis elegans C-terminal binding protein (CtBP) and show that it adopts a fold containing a treble clef motif, bearing similarity to the zinc finger-associated domain (ZAD) from Drosophila Grauzone. The CtBP THAP domain contains a large, positively charged surface patch and we demonstrate that this domain can bind to double-stranded DNA in an electrophoretic mobility-shift assay. These data, together with existing reports, indicate that THAP domains might exhibit a functional diversity similar to that observed for classical and GATA-type zinc fingers. © 2006 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"yu-kong-dore-abdulmalik-katein-zhou-choi-gell-etal-anerythroidchaperonethatfacilitatesfoldingofglobinsubunitsforhemoglobinsynthesis-2007\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  An erythroid chaperone that facilitates folding of α-globin subunits for hemoglobin synthesis.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Yu,  X.; Kong,  Y.; Dore,  L.; Abdulmalik,  O.; Katein,  A.; Zhou,  S.; Choi,  J.; Gell,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Gow,  A.; Gow,  A.;  and Weiss,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Clinical Investigation</i>, 117(7): 1856-1865.  2007.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1172/JCI31664\"\n     onclick=\"log_download('yu-kong-dore-abdulmalik-katein-zhou-choi-gell-etal-anerythroidchaperonethatfacilitatesfoldingofglobinsubunitsforhemoglobinsynthesis-2007', 'http://doi.org/10.1172/JCI31664', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/yu-kong-dore-abdulmalik-katein-zhou-choi-gell-etal-anerythroidchaperonethatfacilitatesfoldingofglobinsubunitsforhemoglobinsynthesis-2007\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('yu-kong-dore-abdulmalik-katein-zhou-choi-gell-etal-anerythroidchaperonethatfacilitatesfoldingofglobinsubunitsforhemoglobinsynthesis-2007')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {An erythroid chaperone that facilitates folding of α-globin subunits for hemoglobin synthesis},\n type = {article},\n year = {2007},\n pages = {1856-1865},\n volume = {117},\n id = {b36c185f-d2dd-356f-b1e4-9fa2eef4f751},\n created = {2023-01-10T01:45:46.881Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:46.881Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Erythrocyte precursors produce abundant α- and β-globin proteins, which assemble with each other to form hemoglobin A (HbA), the major blood oxygen carrier. αHb-stabilizing protein (AHSP) binds free α subunits reversibly to maintain their structure and limit their ability to generate reactive oxygen species. Accordingly, loss of AHSP aggravates the toxicity of excessive free α-globin caused by β-globin gene disruption in mice. Surprisingly, we found that AHSP also has important functions when free α-globin is limited. Thus, compound mutants lacking both Ahsp and 1 of 4 α-globin genes (genotype Ahsp-/-α-globin -/-α/αα) exhibited more severe anemia and Hb instability than mice with either mutation alone. In vitro, recombinant AHSP promoted folding of newly translated α-globin, enhanced its refolding after denaturation, and facilitated its incorporation into HbA. Moreover, in erythroid precursors, newly formed free α-globin was destabilized by loss of AHSP. Therefore, in addition to its previously defined role in detoxification of excess α-globin, AHSP also acts as a molecular chaperone to stabilize nascent α-globin for HbA assembly. Our findings illustrate what we believe to be a novel adaptive mechanism by which a specialized cell coordinates high-level production of a multisubunit protein and protects against various synthetic imbalances.},\n bibtype = {article},\n author = {Yu, X. and Kong, Y. and Dore, L.C. and Abdulmalik, O. and Katein, A.M. and Zhou, S. and Choi, J.K. and Gell, D. and Mackay, J.P. and Gow, A.J. and Gow, A.J. and Weiss, M.J.},\n doi = {10.1172/JCI31664},\n journal = {Journal of Clinical Investigation},\n number = {7}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Erythrocyte precursors produce abundant α- and β-globin proteins, which assemble with each other to form hemoglobin A (HbA), the major blood oxygen carrier. αHb-stabilizing protein (AHSP) binds free α subunits reversibly to maintain their structure and limit their ability to generate reactive oxygen species. Accordingly, loss of AHSP aggravates the toxicity of excessive free α-globin caused by β-globin gene disruption in mice. Surprisingly, we found that AHSP also has important functions when free α-globin is limited. Thus, compound mutants lacking both Ahsp and 1 of 4 α-globin genes (genotype Ahsp-/-α-globin -/-α/αα) exhibited more severe anemia and Hb instability than mice with either mutation alone. In vitro, recombinant AHSP promoted folding of newly translated α-globin, enhanced its refolding after denaturation, and facilitated its incorporation into HbA. Moreover, in erythroid precursors, newly formed free α-globin was destabilized by loss of AHSP. Therefore, in addition to its previously defined role in detoxification of excess α-globin, AHSP also acts as a molecular chaperone to stabilize nascent α-globin for HbA assembly. Our findings illustrate what we believe to be a novel adaptive mechanism by which a specialized cell coordinates high-level production of a multisubunit protein and protects against various synthetic imbalances.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kirk-sunde-costa-rankin-wolstein-castro-butler-hyun-etal-mutationsincrdiactboxfactorgenetbx20areassociatedwithdiversecardiacpathologiesincludingdefectsofseptationandvalvulogenesisandcardiomyopathy-2007\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Mutations in crdiac T-box factor gene TBX20 are associated with diverse cardiac pathologies, including defects of septation and valvulogenesis and cardiomyopathy.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kirk,  E.; Sunde,  M.; Costa,  M.; Rankin,  S.; Wolstein,  O.; Castro,  M.; Butler,  T.; Hyun,  C.; Guo,  G.; Otway,  R.; Winlaw,  D.;  and Harvey,  R.\n</span>\n\n  \n\n</span>\n\n  <i>American Journal of Human Genetics</i>, 81(2): 280-291.  2007.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1086/519530\"\n     onclick=\"log_download('kirk-sunde-costa-rankin-wolstein-castro-butler-hyun-etal-mutationsincrdiactboxfactorgenetbx20areassociatedwithdiversecardiacpathologiesincludingdefectsofseptationandvalvulogenesisandcardiomyopathy-2007', 'http://doi.org/10.1086/519530', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kirk-sunde-costa-rankin-wolstein-castro-butler-hyun-etal-mutationsincrdiactboxfactorgenetbx20areassociatedwithdiversecardiacpathologiesincludingdefectsofseptationandvalvulogenesisandcardiomyopathy-2007\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kirk-sunde-costa-rankin-wolstein-castro-butler-hyun-etal-mutationsincrdiactboxfactorgenetbx20areassociatedwithdiversecardiacpathologiesincludingdefectsofseptationandvalvulogenesisandcardiomyopathy-2007')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Mutations in crdiac T-box factor gene TBX20 are associated with diverse cardiac pathologies, including defects of septation and valvulogenesis and cardiomyopathy},\n type = {article},\n year = {2007},\n pages = {280-291},\n volume = {81},\n id = {52c04b2f-8782-380e-959a-24222788591c},\n created = {2023-01-10T01:45:47.818Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:47.818Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The T-box family transcription factor gene TBX20 acts in a conserved regulatory network, guiding heart formation and patterning in diverse species. Mouse Tbx20 is expressed in cardiac progenitor cells, differentiating cardiomyocytes, and developing valvular tissue, and its deletion or RNA interference-mediated knockdown is catastrophic for heart development. TBX20 interacts physically, functionally, and genetically with other cardiac transcription factors, including NKX2-5, GATA4, and TBX5, mutations of which cause congenital heart disease (CHD). Here, we report nonsense (Q195X) and missense (I152M) germline mutations within the T-box DNA-binding domain of human TBX20 that were associated with a family history of CHD and a complex spectrum of developmental anomalies, including defects in septation, chamber growth, and valvulogenesis. Biophysical characterization of wild-type and mutant proteins indicated how the missense mutation disrupts the structure and function of the TBX20 T-box. Dilated cardiomyopathy was a feature of the TBX20 mutant phenotype in humans and mice, suggesting that mutations in developmental transcription factors can provide a sensitized template for adult-onset heart disease. Our findings are the first to link TBX20 mutations to human pathology. They provide insights into how mutation of different genes in an interactive regulatory circuit lead to diverse clinical phenotypes, with implications for diagnosis, genetic screening, and patient follow-up. © 2007 by The American Society of Human Genetics. All rights reserved.},\n bibtype = {article},\n author = {Kirk, E.P. and Sunde, M. and Costa, M.W. and Rankin, S.A. and Wolstein, O. and Castro, M.L. and Butler, T.L. and Hyun, C. and Guo, G. and Otway, R. and Winlaw, D.S. and Harvey, R.P.},\n doi = {10.1086/519530},\n journal = {American Journal of Human Genetics},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The T-box family transcription factor gene TBX20 acts in a conserved regulatory network, guiding heart formation and patterning in diverse species. Mouse Tbx20 is expressed in cardiac progenitor cells, differentiating cardiomyocytes, and developing valvular tissue, and its deletion or RNA interference-mediated knockdown is catastrophic for heart development. TBX20 interacts physically, functionally, and genetically with other cardiac transcription factors, including NKX2-5, GATA4, and TBX5, mutations of which cause congenital heart disease (CHD). Here, we report nonsense (Q195X) and missense (I152M) germline mutations within the T-box DNA-binding domain of human TBX20 that were associated with a family history of CHD and a complex spectrum of developmental anomalies, including defects in septation, chamber growth, and valvulogenesis. Biophysical characterization of wild-type and mutant proteins indicated how the missense mutation disrupts the structure and function of the TBX20 T-box. Dilated cardiomyopathy was a feature of the TBX20 mutant phenotype in humans and mice, suggesting that mutations in developmental transcription factors can provide a sensitized template for adult-onset heart disease. Our findings are the first to link TBX20 mutations to human pathology. They provide insights into how mutation of different genes in an interactive regulatory circuit lead to diverse clinical phenotypes, with implications for diagnosis, genetic screening, and patient follow-up. © 2007 by The American Society of Human Genetics. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-sunde-lowry-crossley-matthews-proteininteractionsisseeingbelieving-2007\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Protein interactions: is seeing believing?.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Sunde,  M.; Lowry,  J.; Crossley,  M.;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Trends in Biochemical Sciences</i>, 32(12): 530-531.  2007.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.tibs.2007.09.006\"\n     onclick=\"log_download('mackay-sunde-lowry-crossley-matthews-proteininteractionsisseeingbelieving-2007', 'http://doi.org/10.1016/j.tibs.2007.09.006', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-sunde-lowry-crossley-matthews-proteininteractionsisseeingbelieving-2007\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-sunde-lowry-crossley-matthews-proteininteractionsisseeingbelieving-2007')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Protein interactions: is seeing believing?},\n type = {article},\n year = {2007},\n pages = {530-531},\n volume = {32},\n id = {44b87550-332a-36df-aa40-16703ffad894},\n created = {2023-01-10T01:45:48.739Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:48.739Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Mackay, J.P. and Sunde, M. and Lowry, J.A. and Crossley, M. and Matthews, J.M.},\n doi = {10.1016/j.tibs.2007.09.006},\n journal = {Trends in Biochemical Sciences},\n number = {12}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wang-gunar-forwood-teh-catanzariti-lawrence-loughlin-mackay-etal-crystalstructuresofflaxrustavirulenceproteinsavrl567aanddrevealdetailsofthestructuralbasisforflaxdiseaseresistancespecificity-2007\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Crystal structures of flax rust avirulence proteins AvrL567-A and -D reveal details of the structural basis for flax disease resistance specificity.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wang,  C.; Gunčar,  G.; Forwood,  J.; Teh,  T.; Catanzariti,  A.; Lawrence,  G.; Loughlin,  F.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Schirra,  H.; Anderson,  P.; Dodds,  P.;  and Kobe,  B.\n</span>\n\n  \n\n</span>\n\n  <i>Plant Cell</i>, 19(9): 2898-2912.  2007.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1105/tpc.107.053611\"\n     onclick=\"log_download('wang-gunar-forwood-teh-catanzariti-lawrence-loughlin-mackay-etal-crystalstructuresofflaxrustavirulenceproteinsavrl567aanddrevealdetailsofthestructuralbasisforflaxdiseaseresistancespecificity-2007', 'http://doi.org/10.1105/tpc.107.053611', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wang-gunar-forwood-teh-catanzariti-lawrence-loughlin-mackay-etal-crystalstructuresofflaxrustavirulenceproteinsavrl567aanddrevealdetailsofthestructuralbasisforflaxdiseaseresistancespecificity-2007\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wang-gunar-forwood-teh-catanzariti-lawrence-loughlin-mackay-etal-crystalstructuresofflaxrustavirulenceproteinsavrl567aanddrevealdetailsofthestructuralbasisforflaxdiseaseresistancespecificity-2007')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Crystal structures of flax rust avirulence proteins AvrL567-A and -D reveal details of the structural basis for flax disease resistance specificity},\n type = {article},\n year = {2007},\n pages = {2898-2912},\n volume = {19},\n id = {e42e696d-217e-3568-9613-ef60ec204073},\n created = {2023-01-10T01:45:49.665Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:49.665Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The gene-for-gene mechanism of plant disease resistance involves direct or indirect recognition of pathogen avirulence (Avr) proteins by plant resistance (R) proteins. Flax rust (Melampsora lini) AvrL567 avirulence proteins and the corresponding flax (Linum usitatissimum) L5, L6, and L7 resistance proteins interact directly. We determined the three-dimensional structures of two members of the AvrL567 family, AvrL567-A and AvrL567-D, at 1.4- and 2.3-Å resolution, respectively. The structures of both proteins are very similar and reveal a β-sandwich fold with no close known structural homologs. The polymorphic residues in the AvrL567 family map to the surface of the protein, and polymorphisms in residues associated with recognition differences for the R proteins lead to significant changes in surface chemical properties. Analysis of single amino acid substitutions in AvrL567 proteins confirm the role of individual residues in conferring differences in recognition and suggest that the specificity results from the cumulative effects of multiple amino acid contacts. The structures also provide insights into possible pathogen-associated functions of AvrL567 proteins, with nucleic acid binding activity demonstrated in vitro. Our studies provide some of the first structural information on avirulence proteins that bind directly to the corresponding resistance proteins, allowing an examination of the molecular basis of the interaction with the resistance proteins as a step toward designing new resistance specificities. © 2007 American Society of Plant Biologists.},\n bibtype = {article},\n author = {Wang, C.-I.A. and Gunčar, G. and Forwood, J.K. and Teh, T. and Catanzariti, A.-M. and Lawrence, G.J. and Loughlin, F.E. and Mackay, J.P. and Schirra, H.J. and Anderson, P.A. and Dodds, P.N. and Kobe, B.},\n doi = {10.1105/tpc.107.053611},\n journal = {Plant Cell},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The gene-for-gene mechanism of plant disease resistance involves direct or indirect recognition of pathogen avirulence (Avr) proteins by plant resistance (R) proteins. Flax rust (Melampsora lini) AvrL567 avirulence proteins and the corresponding flax (Linum usitatissimum) L5, L6, and L7 resistance proteins interact directly. We determined the three-dimensional structures of two members of the AvrL567 family, AvrL567-A and AvrL567-D, at 1.4- and 2.3-Å resolution, respectively. The structures of both proteins are very similar and reveal a β-sandwich fold with no close known structural homologs. The polymorphic residues in the AvrL567 family map to the surface of the protein, and polymorphisms in residues associated with recognition differences for the R proteins lead to significant changes in surface chemical properties. Analysis of single amino acid substitutions in AvrL567 proteins confirm the role of individual residues in conferring differences in recognition and suggest that the specificity results from the cumulative effects of multiple amino acid contacts. The structures also provide insights into possible pathogen-associated functions of AvrL567 proteins, with nucleic acid binding activity demonstrated in vitro. Our studies provide some of the first structural information on avirulence proteins that bind directly to the corresponding resistance proteins, allowing an examination of the molecular basis of the interaction with the resistance proteins as a step toward designing new resistance specificities. © 2007 American Society of Plant Biologists.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2006\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2006')\"\n      onkeypress=\"toggleGroup('group_2006')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2006</span>\n      <span class=\"bibbase_group_count\">\n        \n        (10)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006', 'http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103', event);\">\n    Structural basis for rodlet assembly in fungal hydrophobins.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H., Y.</a>; Winefield,  R., D.; Sunde,  M.; Matthews,  J., M.; Haverkamp,  R., G.; Templeton,  M., D.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences</i>, 103(10): 3621-3626. 3 2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103\"\n     onclick=\"log_download('kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006', 'http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Structural basis for rodlet assembly in fungal hydrophobins [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.0505704103\"\n     onclick=\"log_download('kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006', 'http://doi.org/10.1073/pnas.0505704103', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural basis for rodlet assembly in fungal hydrophobins},\n type = {article},\n year = {2006},\n keywords = {Amyloid,NMR,Polymer},\n pages = {3621-3626},\n volume = {103},\n websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.0505704103},\n month = {3},\n day = {7},\n id = {5c3390e3-fea1-3491-b3cb-82c763c2d1b5},\n created = {2020-12-17T05:29:58.566Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:58.566Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Kwan2006},\n source_type = {ARTICLE},\n notes = {cited By 150},\n private_publication = {false},\n abstract = {Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.},\n bibtype = {article},\n author = {Kwan, A. H. Y. and Winefield, R. D. and Sunde, M. and Matthews, J. M. and Haverkamp, R. G. and Templeton, M. D. and Mackay, J. P.},\n doi = {10.1073/pnas.0505704103},\n journal = {Proceedings of the National Academy of Sciences},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006', 'https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679', event);\">\n    A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gell,  D., A.; Westman,  B., J.; Gorman,  D.; Liew,  C.; Welch,  J., J.; Weiss,  M., J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 362(2): 287-297. 9 2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679\"\n     onclick=\"log_download('gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006', 'https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2006.07.010\"\n     onclick=\"log_download('gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006', 'http://doi.org/10.1016/j.jmb.2006.07.010', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP},\n type = {article},\n year = {2006},\n keywords = {NMR,haem,haem-binding protein,p22HBP,protein structure},\n pages = {287-297},\n volume = {362},\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S0022283606008679},\n month = {9},\n id = {6eee0455-9c97-35b4-ad59-54f7f318abd4},\n created = {2020-12-17T05:29:58.858Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:58.858Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Gell2006},\n source_type = {ARTICLE},\n notes = {cited By 6},\n private_publication = {false},\n abstract = {The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Gell, David A. and Westman, Belinda J. and Gorman, Daniel and Liew, ChuKong and Welch, John J. and Weiss, Mitchell J. and Mackay, Joel P.},\n doi = {10.1016/j.jmb.2006.07.010},\n journal = {Journal of Molecular Biology},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"lowry-mackay-gata1oneproteinmanypartners-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'lowry-mackay-gata1oneproteinmanypartners-2006', 'https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074', event);\">\n    GATA-1: One protein, many partners.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Lowry,  J., A.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>The International Journal of Biochemistry & Cell Biology</i>, 38(1): 6-11. 1 2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074\"\n     onclick=\"log_download('lowry-mackay-gata1oneproteinmanypartners-2006', 'https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"GATA-1: One protein, many partners [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.biocel.2005.06.017\"\n     onclick=\"log_download('lowry-mackay-gata1oneproteinmanypartners-2006', 'http://doi.org/10.1016/j.biocel.2005.06.017', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/lowry-mackay-gata1oneproteinmanypartners-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('lowry-mackay-gata1oneproteinmanypartners-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {GATA-1: One protein, many partners},\n type = {article},\n year = {2006},\n keywords = {Factor,Finger,Hematopoiesis,Transcription,Zinc},\n pages = {6-11},\n volume = {38},\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S1357272505002074},\n month = {1},\n id = {ef48217b-ded1-36e7-b8a7-8426ac4c459e},\n created = {2020-12-17T05:29:58.983Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:58.983Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Lowry2006},\n source_type = {ARTICLE},\n notes = {cited By 35},\n private_publication = {false},\n abstract = {GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Lowry, Jason A. and Mackay, Joel P.},\n doi = {10.1016/j.biocel.2005.06.017},\n journal = {The International Journal of Biochemistry &amp; Cell Biology},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"loughlin-mackay-zincfingersareknownasdomainsforbindingdnaandrnadotheyalsomediateproteinproteininteractions-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Zinc fingers are known as domains for binding DNA and RNA. Do they also mediate protein-protein interactions?.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Loughlin,  F.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>IUBMB Life</i>, 58(12): 731-733.  2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1080/15216540600868445\"\n     onclick=\"log_download('loughlin-mackay-zincfingersareknownasdomainsforbindingdnaandrnadotheyalsomediateproteinproteininteractions-2006', 'http://doi.org/10.1080/15216540600868445', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/loughlin-mackay-zincfingersareknownasdomainsforbindingdnaandrnadotheyalsomediateproteinproteininteractions-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('loughlin-mackay-zincfingersareknownasdomainsforbindingdnaandrnadotheyalsomediateproteinproteininteractions-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Zinc fingers are known as domains for binding DNA and RNA. Do they also mediate protein-protein interactions?},\n type = {article},\n year = {2006},\n pages = {731-733},\n volume = {58},\n id = {a59a3ace-fadf-3a89-ab6c-424f0813e6d6},\n created = {2023-01-10T01:45:50.577Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:50.577Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Loughlin, F.E. and Mackay, J.P.},\n doi = {10.1080/15216540600868445},\n journal = {IUBMB Life},\n number = {12}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"chu-rand-simpson-yung-mansfield-crossley-prtoriusibba-nerlov-etal-molecularanalysisoftheinteractionbetweenthehematopoieticmastertranscriptionfactorsgata1andpu-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Molecular analysis of the interaction between the hematopoietic master transcription factors GATA-1 and PU.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Chu,  W.; Rand,  K.; Simpson,  R.; Yung,  W.; Mansfield,  R.; Crossley,  M.; Prœtorius-Ibba,  M.; Nerlov,  C.; Poulsen,  F.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 281(38): 28296-28306.  2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M602830200\"\n     onclick=\"log_download('chu-rand-simpson-yung-mansfield-crossley-prtoriusibba-nerlov-etal-molecularanalysisoftheinteractionbetweenthehematopoieticmastertranscriptionfactorsgata1andpu-2006', 'http://doi.org/10.1074/jbc.M602830200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/chu-rand-simpson-yung-mansfield-crossley-prtoriusibba-nerlov-etal-molecularanalysisoftheinteractionbetweenthehematopoieticmastertranscriptionfactorsgata1andpu-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('chu-rand-simpson-yung-mansfield-crossley-prtoriusibba-nerlov-etal-molecularanalysisoftheinteractionbetweenthehematopoieticmastertranscriptionfactorsgata1andpu-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Molecular analysis of the interaction between the hematopoietic master transcription factors GATA-1 and PU},\n type = {article},\n year = {2006},\n pages = {28296-28306},\n volume = {281},\n id = {62d88558-80bb-327f-9a17-f07a26210a6c},\n created = {2023-01-10T01:45:51.530Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:51.530Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {GATA-1 and PU.1 are transcription factors that control erythroid and myeloid development, respectively. The two proteins have been shown to function in an antagonistic fashion, with GATA-1 repressing PU.1 activity during erythropoiesis and PU.1 repressing GATA-1 function during myelopoiesis. It has also become clear that this functional antagonism involves direct interactions between the two proteins. However, the molecular basis for these interactions is not known, and a number of inconsistencies exist in the literature. We have used a range of biophysical methods to define the molecular details of the GATA-1-PU.1 interaction. A combination of NMR titration data and extensive mutagenesis revealed that the PU.1-Ets domain and the GATA-1 C-terminal zinc finger (CF) form a low affinity interaction in which specific regions of each protein are implicated. Surprisingly, the interaction cannot be disrupted by single alanine substitution mutations, suggesting that binding is distributed over an extended interface. The C-terminal basic tail region of CF appears to be sufficient to mediate an interaction with PU.1-Ets, and neither acetylation nor phosphorylation of a peptide corresponding to this region disrupts binding, indicating that the interaction is not dominated by electrostatic interactions. The CF basic tail shares significant sequence homology with the PU.1 interacting motif from c-Jun, suggesting that GATA-1 and c-Jun might compete to bind PU.1. Taken together, our data provide a molecular perspective on the GATA-1-PU.1 interaction, resolving several issues in the existing data and providing insight into the mechanisms through which these two proteins combine to regulate blood development. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Chu, W.L. and Rand, K.D. and Simpson, R.J.Y. and Yung, W.W. and Mansfield, R.E. and Crossley, M. and Prœtorius-Ibba, M. and Nerlov, C. and Poulsen, F.M. and Mackay, J.P.},\n doi = {10.1074/jbc.M602830200},\n journal = {Journal of Biological Chemistry},\n number = {38}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  GATA-1 and PU.1 are transcription factors that control erythroid and myeloid development, respectively. The two proteins have been shown to function in an antagonistic fashion, with GATA-1 repressing PU.1 activity during erythropoiesis and PU.1 repressing GATA-1 function during myelopoiesis. It has also become clear that this functional antagonism involves direct interactions between the two proteins. However, the molecular basis for these interactions is not known, and a number of inconsistencies exist in the literature. We have used a range of biophysical methods to define the molecular details of the GATA-1-PU.1 interaction. A combination of NMR titration data and extensive mutagenesis revealed that the PU.1-Ets domain and the GATA-1 C-terminal zinc finger (CF) form a low affinity interaction in which specific regions of each protein are implicated. Surprisingly, the interaction cannot be disrupted by single alanine substitution mutations, suggesting that binding is distributed over an extended interface. The C-terminal basic tail region of CF appears to be sufficient to mediate an interaction with PU.1-Ets, and neither acetylation nor phosphorylation of a peptide corresponding to this region disrupts binding, indicating that the interaction is not dominated by electrostatic interactions. The CF basic tail shares significant sequence homology with the PU.1 interacting motif from c-Jun, suggesting that GATA-1 and c-Jun might compete to bind PU.1. Taken together, our data provide a molecular perspective on the GATA-1-PU.1 interaction, resolving several issues in the existing data and providing insight into the mechanisms through which these two proteins combine to regulate blood development. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"ryan-sunde-kwan-marianayagam-nancarrow-vandenhoven-thompson-baca-etal-identificationofthekeylmo2bindingdeterminantsonldb1-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Identification of the Key LMO2-binding Determinants on Ldb1.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Ryan,  D.; Sunde,  M.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Marianayagam,  N.; Nancarrow,  A.; vanden Hoven,  R.; Thompson,  L.; Baca,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Visvader,  J.; Visvader,  J.;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 359(1): 66-75.  2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2006.02.074\"\n     onclick=\"log_download('ryan-sunde-kwan-marianayagam-nancarrow-vandenhoven-thompson-baca-etal-identificationofthekeylmo2bindingdeterminantsonldb1-2006', 'http://doi.org/10.1016/j.jmb.2006.02.074', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/ryan-sunde-kwan-marianayagam-nancarrow-vandenhoven-thompson-baca-etal-identificationofthekeylmo2bindingdeterminantsonldb1-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('ryan-sunde-kwan-marianayagam-nancarrow-vandenhoven-thompson-baca-etal-identificationofthekeylmo2bindingdeterminantsonldb1-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Identification of the Key LMO2-binding Determinants on Ldb1},\n type = {article},\n year = {2006},\n pages = {66-75},\n volume = {359},\n id = {14513112-4f7f-308b-b9ed-cb4aa5fd5f5b},\n created = {2023-01-10T01:45:52.515Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:52.515Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The overexpression of LIM-only protein 2 (LMO2) in T-cells, as a result of chromosomal translocations, retroviral insertion during gene therapy, or in transgenic mice models, leads to the onset of T-cell leukemias. LMO2 comprises two protein-binding LIM domains that allow LMO2 to interact with multiple protein partners, including LIM domain-binding protein 1 (Ldb1, also known as CLIM2 and NLI), an essential cofactor for LMO proteins. Sequestration of Ldb1 by LMO2 in T-cells may prevent it binding other key partners, such as LMO4. Here, we show using protein engineering and enzyme-linked immunosorbent assay (ELISA) methodologies that LMO2 binds Ldb1 with a twofold lower affinity than does LMO4. Thus, excess LMO2 rather than an intrinsically higher binding affinity would lead to sequestration of Ldb1. Both LIM domains of LMO2 are required for high-affinity binding to Ldb1 (KD=2.0×10-8 M). However, the first LIM domain of LMO2 is primarily responsible for binding to Ldb1 (KD=2.3×10-7 M), whereas the second LIM domain increases binding by an order of magnitude. We used mutagenesis in combination with yeast two-hybrid analysis, and phage display selection to identify LMO2-binding &quot;hot spots&quot; within Ldb1 that locate to the LIM1-binding region. The delineation of this region reveals some specific differences when compared to the equivalent LMO4:Ldb1 interaction that hold promise for the development of reagents to specifically bind LMO2 in the treatment of leukemia. © 2006 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Ryan, D.P. and Sunde, M. and Kwan, A.H.-Y. and Marianayagam, N.J. and Nancarrow, A.L. and vanden Hoven, R.N. and Thompson, L.S. and Baca, M. and Mackay, J.P. and Visvader, J.E. and Visvader, J.E. and Matthews, J.M.},\n doi = {10.1016/j.jmb.2006.02.074},\n journal = {Journal of Molecular Biology},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The overexpression of LIM-only protein 2 (LMO2) in T-cells, as a result of chromosomal translocations, retroviral insertion during gene therapy, or in transgenic mice models, leads to the onset of T-cell leukemias. LMO2 comprises two protein-binding LIM domains that allow LMO2 to interact with multiple protein partners, including LIM domain-binding protein 1 (Ldb1, also known as CLIM2 and NLI), an essential cofactor for LMO proteins. Sequestration of Ldb1 by LMO2 in T-cells may prevent it binding other key partners, such as LMO4. Here, we show using protein engineering and enzyme-linked immunosorbent assay (ELISA) methodologies that LMO2 binds Ldb1 with a twofold lower affinity than does LMO4. Thus, excess LMO2 rather than an intrinsically higher binding affinity would lead to sequestration of Ldb1. Both LIM domains of LMO2 are required for high-affinity binding to Ldb1 (KD=2.0×10-8 M). However, the first LIM domain of LMO2 is primarily responsible for binding to Ldb1 (KD=2.3×10-7 M), whereas the second LIM domain increases binding by an order of magnitude. We used mutagenesis in combination with yeast two-hybrid analysis, and phage display selection to identify LMO2-binding \"hot spots\" within Ldb1 that locate to the LIM1-binding region. The delineation of this region reveals some specific differences when compared to the equivalent LMO4:Ldb1 interaction that hold promise for the development of reagents to specifically bind LMO2 in the treatment of leukemia. © 2006 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural basis for rodlet assembly in fungal hydrophobins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Winefield,  R.; Sunde,  M.; Matthews,  J.; Haverkamp,  R.; Templeton,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 103(10): 3621-3626.  2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.0505704103\"\n     onclick=\"log_download('kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006', 'http://doi.org/10.1073/pnas.0505704103', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kwan-winefield-sunde-matthews-haverkamp-templeton-mackay-structuralbasisforrodletassemblyinfungalhydrophobins-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural basis for rodlet assembly in fungal hydrophobins},\n type = {article},\n year = {2006},\n pages = {3621-3626},\n volume = {103},\n id = {0944c3ce-33ab-33ae-b13f-43193a7e6f03},\n created = {2023-01-10T01:45:53.429Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:53.429Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.},\n bibtype = {article},\n author = {Kwan, A.H.Y. and Winefield, R.D. and Sunde, M. and Matthews, J.M. and Haverkamp, R.G. and Templeton, M.D. and Mackay, J.P.},\n doi = {10.1073/pnas.0505704103},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Class I hydrophobins are a unique family of fungal proteins that form a polymeric, water-repellent monolayer on the surface of structures such as spores and fruiting bodies. Similar monolayers are being discovered on an increasing range of important microorganisms. Hydrophobin monolayers are amphipathic and particularly robust, and they reverse the wettability of the surface on which they are formed. There are also significant similarities between these polymers and amyloid-like fibrils. However, structural information on these proteins and the rodlets they form has been elusive. Here, we describe the three-dimensional structure of the monomeric form of the class I hydrophobin EAS. EAS forms a β-barrel structure punctuated by several disordered regions and displays a complete segregation of charged and hydrophobic residues on its surface. This structure is consistent with its ability to form an amphipathic polymer. By using this structure, together with data from mutagenesis and previous biophysical studies, we have been able to propose a model for the polymeric rodlet structure adopted by these proteins. X-ray fiber diffraction data from EAS rodlets are consistent with our model. Our data provide molecular insight into the nature of hydrophobin rodlet films and extend our understanding of the fibrillar β-structures that continue to be discovered in the protein world. © 2006 by The National Academy of Sciences of the USA.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kook-yung-simpson-kee-shin-lowry-loughlin-yin-etal-analysisofthestructureandfunctionofthetranscriptionalcoregulatorhop-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Analysis of the structure and function of the transcriptional coregulator HOP.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kook,  H.; Yung,  W.; Simpson,  R.; Kee,  H.; Shin,  S.; Lowry,  J.; Loughlin,  F.; Yin,  Z.; Epstein,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 45(35): 10584-10590.  2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi060641s\"\n     onclick=\"log_download('kook-yung-simpson-kee-shin-lowry-loughlin-yin-etal-analysisofthestructureandfunctionofthetranscriptionalcoregulatorhop-2006', 'http://doi.org/10.1021/bi060641s', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kook-yung-simpson-kee-shin-lowry-loughlin-yin-etal-analysisofthestructureandfunctionofthetranscriptionalcoregulatorhop-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kook-yung-simpson-kee-shin-lowry-loughlin-yin-etal-analysisofthestructureandfunctionofthetranscriptionalcoregulatorhop-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Analysis of the structure and function of the transcriptional coregulator HOP},\n type = {article},\n year = {2006},\n pages = {10584-10590},\n volume = {45},\n id = {ad62d749-5828-3220-9e44-df8440515254},\n created = {2023-01-10T01:45:54.347Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:54.347Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Homeodomain-only protein (HOP) is an 8-kDa transcriptional corepressor that is essential for the normal development of the mammalian heart. Previous studies have shown that HOP, which consists entirely of a putative homeodomain, acts downstream of Nkx2.5 and associates with the serum response factor (SRF), repressing transcription from SRF-responsive genes. HOP is also able to recruit histone deacetylase (HDAC) activity, consistent with its ability to repress transcription. Unlike other classic homeodomain proteins, HOP does not appear to interact with DNA, although it has been unclear if this is because of an overall divergent structure or because of specific amino acid differences between HOP and other homeodomains. To work toward an understanding of HOP function, we have determined the 3D structure of full-length HOP and used a range of biochemical assays to define the parts of the protein that are functionally important for its repression activity. We show that HOP forms a classical homeodomain fold but that it cannot recognize double stranded DNA, a result that emphasizes the importance of caution in predicting protein function from sequence homology alone. We also demonstrate that two distinct regions on the surface of HOP are required for its ability to repress an SRF-driven reporter gene, and it is likely that these motifs direct interactions between HOP and partner proteins such as SRF- and HDAC-containing complexes. Our results demonstrate that the homeodomain fold has been co-opted during evolution for functions other than sequence-specific DNA binding and suggest that HOP functions as an adaptor protein to mediate transcriptional repression. © 2006 American Chemical Society.},\n bibtype = {article},\n author = {Kook, H. and Yung, W.W. and Simpson, R.J. and Kee, H.J. and Shin, S. and Lowry, J.A. and Loughlin, F.E. and Yin, Z. and Epstein, J.A. and Mackay, J.P.},\n doi = {10.1021/bi060641s},\n journal = {Biochemistry},\n number = {35}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Homeodomain-only protein (HOP) is an 8-kDa transcriptional corepressor that is essential for the normal development of the mammalian heart. Previous studies have shown that HOP, which consists entirely of a putative homeodomain, acts downstream of Nkx2.5 and associates with the serum response factor (SRF), repressing transcription from SRF-responsive genes. HOP is also able to recruit histone deacetylase (HDAC) activity, consistent with its ability to repress transcription. Unlike other classic homeodomain proteins, HOP does not appear to interact with DNA, although it has been unclear if this is because of an overall divergent structure or because of specific amino acid differences between HOP and other homeodomains. To work toward an understanding of HOP function, we have determined the 3D structure of full-length HOP and used a range of biochemical assays to define the parts of the protein that are functionally important for its repression activity. We show that HOP forms a classical homeodomain fold but that it cannot recognize double stranded DNA, a result that emphasizes the importance of caution in predicting protein function from sequence homology alone. We also demonstrate that two distinct regions on the surface of HOP are required for its ability to repress an SRF-driven reporter gene, and it is likely that these motifs direct interactions between HOP and partner proteins such as SRF- and HDAC-containing complexes. Our results demonstrate that the homeodomain fold has been co-opted during evolution for functions other than sequence-specific DNA binding and suggest that HOP functions as an adaptor protein to mediate transcriptional repression. © 2006 American Chemical Society.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gell,  D.; Westman,  B.; Gorman,  D.; Liew,  C.; Welch,  J.; Weiss,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Molecular Biology</i>, 362(2): 287-297.  2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jmb.2006.07.010\"\n     onclick=\"log_download('gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006', 'http://doi.org/10.1016/j.jmb.2006.07.010', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gell-westman-gorman-liew-welch-weiss-mackay-anovelhaembindinginterfaceinthe22kdahaembindingproteinp22hbp-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A Novel Haem-binding Interface in the 22 kDa Haem-binding Protein p22HBP},\n type = {article},\n year = {2006},\n pages = {287-297},\n volume = {362},\n id = {f66fd98b-e46d-356c-adc5-dbb7bb45a1c1},\n created = {2023-01-10T01:45:55.259Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:55.259Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Gell, D.A. and Westman, B.J. and Gorman, D. and Liew, C. and Welch, J.J. and Weiss, M.J. and Mackay, J.P.},\n doi = {10.1016/j.jmb.2006.07.010},\n journal = {Journal of Molecular Biology},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The 22 kDa haem-binding protein, p22HBP, is highly expressed in erythropoietic tissues and binds to a range of metallo- and non-metalloporphyrin molecules with similar affinities, suggesting a role in haem regulation or synthesis. We have determined the three-dimensional solution structure of p22HBP and mapped the porphyrin-binding site, which comprises a number of loops and a α-helix all located on a single face of the molecule. The structure of p22HBP is related to the bacterial multi-drug resistance protein BmrR, and is the first protein with this fold to be identified in eukaryotes. Strikingly, the porphyrin-binding site in p22HBP is located in a similar position to the drug-binding site of BmrR. These similarities suggest that the broad ligand specificity observed for both BmrR and p22HBP may result from a conserved ligand interaction mechanism. Taken together, these data suggest that the both the fold and its associated function, that of binding to a broad range of small hydrophobic molecules, are ancient, and have been adapted throughout evolution for a variety of purposes. © 2006 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"lowry-mackay-gata1oneproteinmanypartners-2006\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  GATA-1: One protein, many partners.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Lowry,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>International Journal of Biochemistry and Cell Biology</i>, 38(1): 6-11.  2006.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.biocel.2005.06.017\"\n     onclick=\"log_download('lowry-mackay-gata1oneproteinmanypartners-2006', 'http://doi.org/10.1016/j.biocel.2005.06.017', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/lowry-mackay-gata1oneproteinmanypartners-2006\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('lowry-mackay-gata1oneproteinmanypartners-2006')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {GATA-1: One protein, many partners},\n type = {article},\n year = {2006},\n pages = {6-11},\n volume = {38},\n id = {91c99222-cb02-3632-8751-7f2b89a9578c},\n created = {2023-01-10T01:45:56.163Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:56.163Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Lowry, J.A. and MacKay, J.P.},\n doi = {10.1016/j.biocel.2005.06.017},\n journal = {International Journal of Biochemistry and Cell Biology},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  GATA-1, the founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocytic lineages. GATA-1 regulates the expression of many genes within these lineages and its functionality depends upon its ability to bind both DNA and protein partners. Disruption of either of these functions causes severe hematopoietic dysfunction and results in blood disorders, such as thrombocytopenia and anemia. Within this review, we will focus on the structural aspects of GATA-1 with regard to interactions with its many partners and the identification of several mutations that disrupt these interactions. © 2005 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2005\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2005')\"\n      onkeypress=\"toggleGroup('group_2005')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2005</span>\n      <span class=\"bibbase_group_count\">\n        \n        (12)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/15644435\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005', 'http://www.ncbi.nlm.nih.gov/pubmed/15644435', event);\">\n    Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/Friend of GATA interaction.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Liew,  C., K.; Simpson,  R., J., Y.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A., H., Y.</a>; Crofts,  L., A.; Loughlin,  F., E.; Matthews,  J., M.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences</i>, 102(3): 583-588. 1 2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/15644435\"\n     onclick=\"log_download('liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005', 'http://www.ncbi.nlm.nih.gov/pubmed/15644435', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/Friend of GATA interaction [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.0407511102\"\n     onclick=\"log_download('liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005', 'http://doi.org/10.1073/pnas.0407511102', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/Friend of GATA interaction},\n type = {article},\n year = {2005},\n keywords = {Factor,Gene expression,Transcription},\n pages = {583-588},\n volume = {102},\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/15644435},\n month = {1},\n day = {18},\n id = {4fa93b4d-9743-397c-8bc8-7dec216c8aa7},\n created = {2020-12-17T05:29:53.577Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:53.577Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Liew2005},\n source_type = {ARTICLE},\n notes = {cited By 65},\n private_publication = {false},\n abstract = {GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.},\n bibtype = {article},\n author = {Liew, Chu Kong and Simpson, Raina J Y and Kwan, Ann H Y and Crofts, Linda A. and Loughlin, Fionna E. and Matthews, Jacqueline M. and Crossley, Merlin and Mackay, Joel P.},\n doi = {10.1073/pnas.0407511102},\n journal = {Proceedings of the National Academy of Sciences},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/15841400\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005', 'http://www.ncbi.nlm.nih.gov/pubmed/15841400', event);\">\n    Grb7-SH2 domain dimerisation is affected by a single point mutation.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Porter,  C., J.; Wilce,  M., C., J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Leedman,  P.;  and Wilce,  J., A.\n</span>\n\n  \n\n</span>\n\n  <i>European biophysics journal : EBJ</i>, 34(5): 454-60. 7 2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/15841400\"\n     onclick=\"log_download('porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005', 'http://www.ncbi.nlm.nih.gov/pubmed/15841400', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Grb7-SH2 domain dimerisation is affected by a single point mutation. [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s00249-005-0480-1\"\n     onclick=\"log_download('porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005', 'http://doi.org/10.1007/s00249-005-0480-1', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Grb7-SH2 domain dimerisation is affected by a single point mutation.},\n type = {article},\n year = {2005},\n keywords = {Analytical ultracentrifugation,Growth factor receptor bound protein 7,Protein engineering,Size-exclusion chromatography,Src homology 2 domain},\n pages = {454-60},\n volume = {34},\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/15841400},\n month = {7},\n day = {20},\n id = {042bb7c3-136a-3d03-a241-4b3ebfa7ee45},\n created = {2020-12-17T05:29:56.590Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:56.590Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Porter2005},\n source_type = {ARTICLE},\n notes = {cited By 20},\n private_publication = {false},\n abstract = {Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 muM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation.},\n bibtype = {article},\n author = {Porter, Corrine J. and Wilce, Matthew C J and Mackay, Joel P. and Leedman, Peter and Wilce, Jackie A.},\n doi = {10.1007/s00249-005-0480-1},\n journal = {European biophysics journal : EBJ},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 muM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Grb7-SH2 domain dimerisation is affected by a single point mutation.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Porter,  C.; Wilce,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Leedman,  P.;  and Wilce,  J.\n</span>\n\n  \n\n</span>\n\n  <i>European Biophysics Journal</i>, 34(5): 454-460.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s00249-005-0480-1\"\n     onclick=\"log_download('porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005', 'http://doi.org/10.1007/s00249-005-0480-1', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('porter-wilce-mackay-leedman-wilce-grb7sh2domaindimerisationisaffectedbyasinglepointmutation-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Grb7-SH2 domain dimerisation is affected by a single point mutation},\n type = {article},\n year = {2005},\n pages = {454-460},\n volume = {34},\n id = {273a7a2d-ac31-3984-b4ee-f7adf13fb08c},\n created = {2023-01-10T01:45:57.071Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:57.071Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 μM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation. © EBSA 2005.},\n bibtype = {article},\n author = {Porter, C.J. and Wilce, M.C.J. and Mackay, J.P. and Leedman, P. and Wilce, J.A.},\n doi = {10.1007/s00249-005-0480-1},\n journal = {European Biophysics Journal},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 μM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation. © EBSA 2005.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"feng-zhou-gu-gell-mackay-weiss-gow-shi-structureofoxidizedhaemoglobinboundtoahsprevealsaprotectivemechanismforhaem-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structure of oxidized α-haemoglobin bound to AHSP reveals a protective mechanism for haem.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Feng,  L.; Zhou,  S.; Gu,  L.; Gell,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Weiss,  M.; Gow,  A.;  and Shi,  Y.\n</span>\n\n  \n\n</span>\n\n  <i>Nature</i>, 435(7042): 697-701.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/nature03609\"\n     onclick=\"log_download('feng-zhou-gu-gell-mackay-weiss-gow-shi-structureofoxidizedhaemoglobinboundtoahsprevealsaprotectivemechanismforhaem-2005', 'http://doi.org/10.1038/nature03609', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/feng-zhou-gu-gell-mackay-weiss-gow-shi-structureofoxidizedhaemoglobinboundtoahsprevealsaprotectivemechanismforhaem-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('feng-zhou-gu-gell-mackay-weiss-gow-shi-structureofoxidizedhaemoglobinboundtoahsprevealsaprotectivemechanismforhaem-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structure of oxidized α-haemoglobin bound to AHSP reveals a protective mechanism for haem},\n type = {article},\n year = {2005},\n pages = {697-701},\n volume = {435},\n id = {a2006ec4-0c0d-3521-8be7-b22dddf9fea3},\n created = {2023-01-10T01:45:57.993Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:57.993Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The synthesis of haemoglobin A (HbA) is exquisitely coordinated during erythrocyte development to prevent damaging effects from individual α- and β-subunits. The α-haemoglobin-stabilizing protein (AHSP) binds α-haemoglobin (αHb), inhibits the ability of αHb to generate reactive oxygen species and prevents its precipitation on exposure to oxidant stress. The structure of AHSP bound to ferrous αHb is thought to represent a transitional complex through which αHb is converted to a non-reactive, hexacoordinate ferric form. Here we report the crystal structure of this ferric αHb-AHSP complex at 2.4 A resolution. Our findings reveal a striking bis-histidyl configuration in which both the proximal and the distal histidines coordinate the haem iron atom. To attain this unusual conformation, segments of αHb undergo drastic structural rearrangements, including the repositioning of several α-helices. Moreover, conversion to the ferric bishistidine configuration strongly and specifically inhibits redox chemistry catalysis and haem loss from αHb. The observed structural changes, which impair the chemical reactivity of haem iron, explain how AHSP stabilizes αHb and prevents its damaging effects in cells.},\n bibtype = {article},\n author = {Feng, L. and Zhou, S. and Gu, L. and Gell, D.A. and Mackay, J.P. and Weiss, M.J. and Gow, A.J. and Shi, Y.},\n doi = {10.1038/nature03609},\n journal = {Nature},\n number = {7042}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The synthesis of haemoglobin A (HbA) is exquisitely coordinated during erythrocyte development to prevent damaging effects from individual α- and β-subunits. The α-haemoglobin-stabilizing protein (AHSP) binds α-haemoglobin (αHb), inhibits the ability of αHb to generate reactive oxygen species and prevents its precipitation on exposure to oxidant stress. The structure of AHSP bound to ferrous αHb is thought to represent a transitional complex through which αHb is converted to a non-reactive, hexacoordinate ferric form. Here we report the crystal structure of this ferric αHb-AHSP complex at 2.4 A resolution. Our findings reveal a striking bis-histidyl configuration in which both the proximal and the distal histidines coordinate the haem iron atom. To attain this unusual conformation, segments of αHb undergo drastic structural rearrangements, including the repositioning of several α-helices. Moreover, conversion to the ferric bishistidine configuration strongly and specifically inhibits redox chemistry catalysis and haem loss from αHb. The observed structural changes, which impair the chemical reactivity of haem iron, explain how AHSP stabilizes αHb and prevents its damaging effects in cells.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"lee-nancarrow-bach-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlhx3ldb1complex4-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of an intramolecular Lhx3:ldb1 complex [4].\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Lee,  C.; Nancarrow,  A.; Bach,  I.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biomolecular NMR</i>, 33(3): 198.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/s10858-005-3209-7\"\n     onclick=\"log_download('lee-nancarrow-bach-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlhx3ldb1complex4-2005', 'http://doi.org/10.1007/s10858-005-3209-7', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/lee-nancarrow-bach-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlhx3ldb1complex4-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('lee-nancarrow-bach-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofanintramolecularlhx3ldb1complex4-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {&lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;15&lt;/sup&gt;N and &lt;sup&gt;13&lt;/sup&gt;C assignments of an intramolecular Lhx3:ldb1 complex [4]},\n type = {article},\n year = {2005},\n pages = {198},\n volume = {33},\n id = {f9aa56cf-83b9-3151-a514-a77d7bddc008},\n created = {2023-01-10T01:45:58.923Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:58.923Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Lee, C. and Nancarrow, A.L. and Bach, I. and Mackay, J.P. and Matthews, J.M.},\n doi = {10.1007/s10858-005-3209-7},\n journal = {Journal of Biomolecular NMR},\n number = {3}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/friend of GATA interaction.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Liew,  C.; Simpson,  R.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Crofts,  L.; Loughlin,  F.; Matthews,  J.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 102(3): 583-588.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.0407511102\"\n     onclick=\"log_download('liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005', 'http://doi.org/10.1073/pnas.0407511102', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('liew-simpson-kwan-crofts-loughlin-matthews-crossley-mackay-zincfingersasproteinrecognitionmotifsstructuralbasisforthegata1friendofgatainteraction-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Zinc fingers as protein recognition motifs: Structural basis for the GATA-1/friend of GATA interaction},\n type = {article},\n year = {2005},\n pages = {583-588},\n volume = {102},\n id = {6ca8effa-53df-38c4-a21a-ded042ec2309},\n created = {2023-01-10T01:45:59.921Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:45:59.921Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.},\n bibtype = {article},\n author = {Liew, C.K. and Simpson, R.J.Y. and Kwan, A.H.Y. and Crofts, L.A. and Loughlin, F.E. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},\n doi = {10.1073/pnas.0407511102},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  GATA-1 and friend of GATA (FOG) are zinc-finger transcription factors that physically interact to play essential roles in erythroid and megakaryocytic development. Several naturally occurring mutations in the GATA-1 gene that alter the FOG-binding domain have been reported. The mutations are associated with familial anemias and thrombocytopenias of differing severity. To elucidate the molecular basis for the GATA-1/FOG interaction, we have determined the three-dimensional structure of a complex comprising the interaction domains of these proteins. The structure reveals how zinc fingers can act as protein recognition motifs. Details of the architecture of the contact domains and their physical properties provide a molecular explanation for how the GATA-1 mutations contribute to distinct but related genetic diseases.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"sharpe-liew-kwan-wilce-crossley-matthews-mackay-assessmentoftherobustnessofaserendipitouszincbindingfoldmutagenesisandproteingrafting-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Assessment of the robustness of a serendipitous zinc binding fold: Mutagenesis and protein grafting.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Sharpe,  B.; Liew,  C.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Wilce,  J.; Crossley,  M.; Matthews,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 13(2): 257-266.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.str.2004.12.007\"\n     onclick=\"log_download('sharpe-liew-kwan-wilce-crossley-matthews-mackay-assessmentoftherobustnessofaserendipitouszincbindingfoldmutagenesisandproteingrafting-2005', 'http://doi.org/10.1016/j.str.2004.12.007', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/sharpe-liew-kwan-wilce-crossley-matthews-mackay-assessmentoftherobustnessofaserendipitouszincbindingfoldmutagenesisandproteingrafting-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('sharpe-liew-kwan-wilce-crossley-matthews-mackay-assessmentoftherobustnessofaserendipitouszincbindingfoldmutagenesisandproteingrafting-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Assessment of the robustness of a serendipitous zinc binding fold: Mutagenesis and protein grafting},\n type = {article},\n year = {2005},\n pages = {257-266},\n volume = {13},\n id = {9aeeb5d0-0dee-3dab-93d2-266c720ce66c},\n created = {2023-01-10T01:46:00.845Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:00.845Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Zinc binding motifs have received much attention in the area of protein design. Here, we have tested the suitability of a recently discovered nonnative zinc binding structure as a protein design scaffold. A series of multiple alanine mutants was created to investigate the minimal requirements for folding, and solution structures of these mutants showed that the original fold was maintained, despite changes in 50% of the sequence. We next attempted to transplant binding faces from chosen bimolecular interactions onto one of these mutants, and many of the resulting &quot;chimeras&quot; were shown to adopt a native-like fold. These results both highlight the robust nature of small zinc binding domains and underscore the complexity of designing functional proteins, even using such small, highly ordered scaffolds as templates.},\n bibtype = {article},\n author = {Sharpe, B.K. and Liew, C.K. and Kwan, A.H. and Wilce, J.A. and Crossley, M. and Matthews, J.M. and Mackay, J.P.},\n doi = {10.1016/j.str.2004.12.007},\n journal = {Structure},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Zinc binding motifs have received much attention in the area of protein design. Here, we have tested the suitability of a recently discovered nonnative zinc binding structure as a protein design scaffold. A series of multiple alanine mutants was created to investigate the minimal requirements for folding, and solution structures of these mutants showed that the original fold was maintained, despite changes in 50% of the sequence. We next attempted to transplant binding faces from chosen bimolecular interactions onto one of these mutants, and many of the resulting \"chimeras\" were shown to adopt a native-like fold. These results both highlight the robust nature of small zinc binding domains and underscore the complexity of designing functional proteins, even using such small, highly ordered scaffolds as templates.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kwan-fairley-anderberg-liew-harding-mackay-solutionstructureofarecombinanttypeisculpinantifreezeprotein-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Solution structure of a recombinant type I sculpin antifreeze protein.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Fairley,  K.; Anderberg,  P.; Liew,  C.; Harding,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 44(6): 1980-1988.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi047782j\"\n     onclick=\"log_download('kwan-fairley-anderberg-liew-harding-mackay-solutionstructureofarecombinanttypeisculpinantifreezeprotein-2005', 'http://doi.org/10.1021/bi047782j', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kwan-fairley-anderberg-liew-harding-mackay-solutionstructureofarecombinanttypeisculpinantifreezeprotein-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kwan-fairley-anderberg-liew-harding-mackay-solutionstructureofarecombinanttypeisculpinantifreezeprotein-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution structure of a recombinant type I sculpin antifreeze protein},\n type = {article},\n year = {2005},\n pages = {1980-1988},\n volume = {44},\n id = {02c3451d-5f79-3362-9ed9-2542509ffb84},\n created = {2023-01-10T01:46:01.762Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:01.762Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {We have determined the solution structure of rSS3, a recombinant form of the type I shorthorn sculpin antifreeze protein (AFP), at 278 and 268 K. This AFP contains an unusual sequence of N-terminal residues, together with two of the 11-residue repeats that are characteristic of the type I winter flounder AFP. The solution conformation of the N-terminal region of the sculpin AFP has been assumed to be the critical factor that results in recognition of different ice planes by the sculpin and flounder AFPs. At 278 K, the two repeats units (residues 11-20 and 21-32) in rSS3 form a continuous α-helix, with the residues 30-33 in the second repeat somewhat less well defined. Within the N-terminal region, residues 2-6 are well defined and helical and linked to the main helix by a more flexible region comprising residues A7-T11. At 268 K the AFP is overall more helical but retains the apparent hinge region. The helical conformation of the two repeats units is almost identical to the corresponding repeats in the type I winter flounder AFP. We also show that while tetracetylated rSS3 has antifreeze activity comparable to the natural AFP, its overall structure is the same as that of the unacetylated peptide. These data provide some insight into the structural determinants of antifreeze activity and should assist in the development of models that explain the recognition of different ice interfaces by the sculpin and flounder type I AFPs.},\n bibtype = {article},\n author = {Kwan, A.H.-Y. and Fairley, K. and Anderberg, P.I. and Liew, C.W. and Harding, M.M. and Mackay, J.P.},\n doi = {10.1021/bi047782j},\n journal = {Biochemistry},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  We have determined the solution structure of rSS3, a recombinant form of the type I shorthorn sculpin antifreeze protein (AFP), at 278 and 268 K. This AFP contains an unusual sequence of N-terminal residues, together with two of the 11-residue repeats that are characteristic of the type I winter flounder AFP. The solution conformation of the N-terminal region of the sculpin AFP has been assumed to be the critical factor that results in recognition of different ice planes by the sculpin and flounder AFPs. At 278 K, the two repeats units (residues 11-20 and 21-32) in rSS3 form a continuous α-helix, with the residues 30-33 in the second repeat somewhat less well defined. Within the N-terminal region, residues 2-6 are well defined and helical and linked to the main helix by a more flexible region comprising residues A7-T11. At 268 K the AFP is overall more helical but retains the apparent hinge region. The helical conformation of the two repeats units is almost identical to the corresponding repeats in the type I winter flounder AFP. We also show that while tetracetylated rSS3 has antifreeze activity comparable to the natural AFP, its overall structure is the same as that of the unacetylated peptide. These data provide some insight into the structural determinants of antifreeze activity and should assist in the development of models that explain the recognition of different ice interfaces by the sculpin and flounder type I AFPs.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"weiss-zhou-feng-gell-mackay-shi-gow-roleofalphahemoglobinstabilizingproteininnormalerythropoiesisandthalassemia-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Role of alpha hemoglobin-stabilizing protein in normal erythropoiesis and β-thalassemia.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Weiss,  M.; Zhou,  S.; Feng,  L.; Gell,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Shi,  Y.;  and Gow,  A.\n</span>\n\n  \n\n</span>\n\n  Volume 1054  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1196/annals.1345.013\"\n     onclick=\"log_download('weiss-zhou-feng-gell-mackay-shi-gow-roleofalphahemoglobinstabilizingproteininnormalerythropoiesisandthalassemia-2005', 'http://doi.org/10.1196/annals.1345.013', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/weiss-zhou-feng-gell-mackay-shi-gow-roleofalphahemoglobinstabilizingproteininnormalerythropoiesisandthalassemia-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('weiss-zhou-feng-gell-mackay-shi-gow-roleofalphahemoglobinstabilizingproteininnormalerythropoiesisandthalassemia-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@book{\n title = {Role of alpha hemoglobin-stabilizing protein in normal erythropoiesis and β-thalassemia},\n type = {book},\n year = {2005},\n source = {Annals of the New York Academy of Sciences},\n pages = {103-117},\n volume = {1054},\n id = {3eadb043-bae4-3de9-9a30-64811004299d},\n created = {2023-01-10T01:46:02.725Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:02.725Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Hemoglobin (Hb) synthesis is coordinated by homeostatic mechanisms to limit the accumulation of free α or β subunits, which are cytotoxic. Alpha hemoglobin-stabilizing protein (AHSP) is an abundant erythroid protein that specifically binds free αHb, stabilizes its structure, and limits its ability to participate in chemical reactions that generate reactive oxygen species. Gene ablation studies in mice demonstrate that AHSP is required for normal erythropoiesis. AHSP-null erythrocytes are short-lived, contain Hb precipitates, and exhibit signs of oxidative damage. Loss of AHSP exacerbates β-thalassemia in mice, indicating that altered AHSP expression or function could modify thalassemia phenotypes in humans, a topic that is beginning to be explored in clinical studies. We used biochemical, spectroscopic, and crystallographic methods to examine how AHSP stabilizes αHb. AHSP binds the G and H helices of αHb on a surface that largely overlaps with the α1-β1 interface of HbA. This result explains previous findings that βHb can competitively displace AHSP from αHb to form HbA tetramer. Remarkably, binding of AHSP to oxygenated αHb induces dramatic conformational changes and converts the heme-bound iron to an oxidized hemichrome state in which all six coordinate positions are occupied. This structure limits the reactivity of heme iron, providing a mechanism by which AHSP stabilizes αHb. These findings suggest a biochemical pathway through which AHSP might participate in normal Hb synthesis and modulate the severity of thalassemias. Moreover, understanding how AHSP stabilizes αHb provides a theoretical basis for new strategies to inhibit the damaging effects of free αHb that accumulates in β-thalassemia. © 2005 New York Academy of Sciences.},\n bibtype = {book},\n author = {Weiss, M.J. and Zhou, S. and Feng, L. and Gell, D.A. and Mackay, J.P. and Shi, Y. and Gow, A.J.},\n doi = {10.1196/annals.1345.013}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Hemoglobin (Hb) synthesis is coordinated by homeostatic mechanisms to limit the accumulation of free α or β subunits, which are cytotoxic. Alpha hemoglobin-stabilizing protein (AHSP) is an abundant erythroid protein that specifically binds free αHb, stabilizes its structure, and limits its ability to participate in chemical reactions that generate reactive oxygen species. Gene ablation studies in mice demonstrate that AHSP is required for normal erythropoiesis. AHSP-null erythrocytes are short-lived, contain Hb precipitates, and exhibit signs of oxidative damage. Loss of AHSP exacerbates β-thalassemia in mice, indicating that altered AHSP expression or function could modify thalassemia phenotypes in humans, a topic that is beginning to be explored in clinical studies. We used biochemical, spectroscopic, and crystallographic methods to examine how AHSP stabilizes αHb. AHSP binds the G and H helices of αHb on a surface that largely overlaps with the α1-β1 interface of HbA. This result explains previous findings that βHb can competitively displace AHSP from αHb to form HbA tetramer. Remarkably, binding of AHSP to oxygenated αHb induces dramatic conformational changes and converts the heme-bound iron to an oxidized hemichrome state in which all six coordinate positions are occupied. This structure limits the reactivity of heme iron, providing a mechanism by which AHSP stabilizes αHb. These findings suggest a biochemical pathway through which AHSP might participate in normal Hb synthesis and modulate the severity of thalassemias. Moreover, understanding how AHSP stabilizes αHb provides a theoretical basis for new strategies to inhibit the damaging effects of free αHb that accumulates in β-thalassemia. © 2005 New York Academy of Sciences.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-muiznieks-toonkool-weiss-thehydrophobicdomain26ofhumantropoelastinisunstructuredinsolution-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The hydrophobic domain 26 of human tropoelastin is unstructured in solution.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Muiznieks,  L.; Toonkool,  P.;  and Weiss,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Structural Biology</i>, 150(2): 154-162.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jsb.2005.02.005\"\n     onclick=\"log_download('mackay-muiznieks-toonkool-weiss-thehydrophobicdomain26ofhumantropoelastinisunstructuredinsolution-2005', 'http://doi.org/10.1016/j.jsb.2005.02.005', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-muiznieks-toonkool-weiss-thehydrophobicdomain26ofhumantropoelastinisunstructuredinsolution-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-muiznieks-toonkool-weiss-thehydrophobicdomain26ofhumantropoelastinisunstructuredinsolution-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The hydrophobic domain 26 of human tropoelastin is unstructured in solution},\n type = {article},\n year = {2005},\n pages = {154-162},\n volume = {150},\n id = {3e2d6e2e-f905-3566-81be-3cc4ef02fb5c},\n created = {2023-01-10T01:46:03.625Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:03.625Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Elastin is the protein responsible for the elastic properties of vertebrate tissue. Very little is currently known about the structure of elastin or of its soluble precursor tropoelastin. We have used high-resolution solution NMR methods to probe the conformational preferences of a conserved hydrophobic region in tropoelastin, domain 26 (D26). Using a combination of homonuclear, 15N-separated and triple resonance experiments, we have obtained essentially full chemical shift assignments for D26 at 278 K. An analysis of secondary chemical shift changes, as well as NOE and 15N relaxation data, leads us to conclude that this domain is essentially unstructured in solution and does not interact with intact tropoelastin. D26 does not display exposed hydrophobic clusters, as expected for a fully unfolded protein and commensurate with an absence of flexible structural motifs, as identified by lack of binding of the fluorescent probe 4,4′-dianilino-1,1′- binaphthyl-5,5′-disulfonic acid. Sedimentation equilibrium data establish that this domain is strictly monomeric in solution. NMR spectra recorded at 278 and 308 K indicate that no significant structural changes occur for this domain over the temperature range 278-308 K, in contrast to the characteristic coacervation behavior that is observed for the full-length protein. © 2005 Elsevier Inc. All rights reserved.},\n bibtype = {article},\n author = {MacKay, J.P. and Muiznieks, L.D. and Toonkool, P. and Weiss, A.S.},\n doi = {10.1016/j.jsb.2005.02.005},\n journal = {Journal of Structural Biology},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Elastin is the protein responsible for the elastic properties of vertebrate tissue. Very little is currently known about the structure of elastin or of its soluble precursor tropoelastin. We have used high-resolution solution NMR methods to probe the conformational preferences of a conserved hydrophobic region in tropoelastin, domain 26 (D26). Using a combination of homonuclear, 15N-separated and triple resonance experiments, we have obtained essentially full chemical shift assignments for D26 at 278 K. An analysis of secondary chemical shift changes, as well as NOE and 15N relaxation data, leads us to conclude that this domain is essentially unstructured in solution and does not interact with intact tropoelastin. D26 does not display exposed hydrophobic clusters, as expected for a fully unfolded protein and commensurate with an absence of flexible structural motifs, as identified by lack of binding of the fluorescent probe 4,4′-dianilino-1,1′- binaphthyl-5,5′-disulfonic acid. Sedimentation equilibrium data establish that this domain is strictly monomeric in solution. NMR spectra recorded at 278 and 308 K indicate that no significant structural changes occur for this domain over the temperature range 278-308 K, in contrast to the characteristic coacervation behavior that is observed for the full-length protein. © 2005 Elsevier Inc. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"duggin-matthews-dixon-wake-mackay-acomplexmechanismdeterminespolarityofdnareplicationforkarrestbythereplicationterminatorcomplexofbacillussubtilis-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A complex mechanism determines polarity of DNA replication fork arrest by the replication terminator complex of Bacillus subtilis.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Duggin,  I.; Matthews,  J.; Dixon,  N.; Wake,  R.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 280(13): 13105-13113.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M414187200\"\n     onclick=\"log_download('duggin-matthews-dixon-wake-mackay-acomplexmechanismdeterminespolarityofdnareplicationforkarrestbythereplicationterminatorcomplexofbacillussubtilis-2005', 'http://doi.org/10.1074/jbc.M414187200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/duggin-matthews-dixon-wake-mackay-acomplexmechanismdeterminespolarityofdnareplicationforkarrestbythereplicationterminatorcomplexofbacillussubtilis-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('duggin-matthews-dixon-wake-mackay-acomplexmechanismdeterminespolarityofdnareplicationforkarrestbythereplicationterminatorcomplexofbacillussubtilis-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A complex mechanism determines polarity of DNA replication fork arrest by the replication terminator complex of Bacillus subtilis},\n type = {article},\n year = {2005},\n pages = {13105-13113},\n volume = {280},\n id = {826a04da-c81b-3dcb-bff3-7b7d2e7fede1},\n created = {2023-01-10T01:46:04.549Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:04.549Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Two dimers of the replication terminator protein (RTP) of Bacillus subtilis bind to a chromosomal DNA terminator site to effect polar replication fork arrest. Cooperative binding of the dimers to overlapping half-sites within the terminator is essential for arrest. It was suggested previously that polarity of fork arrest is the result of the RTP dimer at the blocking (proximal) side within the complex binding very tightly and the permissive-side RTP dimer binding relatively weakly. In order to investigate this &quot;differential binding affinity&quot; model, we have constructed a series of mutant terminators that contain half-sites of widely different RTP binding affinities in various combinations. Although there appeared to be a correlation between binding affinity at the proximal half-site and fork arrest efficiency in vivo for some terminators, several deviated significantly from this correlation. Some terminators exhibited greatly reduced binding cooperativity (and therefore have reduced affinity at each half-site) but were highly efficient in fork arrest, whereas one terminator had normal affinity over the proximal half-site, yet had low fork arrest efficiency. The results show clearly that there is no direct correlation between the RTP binding affinity (either within the full complex or at the proximal half-site within the full complex) and the efficiency of replication fork arrest in vivo. Thus, the differential binding affinity over the proximal and distal half-sites cannot be solely responsible for functional polarity of fork arrest. Furthermore, efficient fork arrest relies on features in addition to the tight binding of RTP to terminator DNA. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc.},\n bibtype = {article},\n author = {Duggin, I.G. and Matthews, J.M. and Dixon, N.E. and Wake, R.G. and Mackay, J.P.},\n doi = {10.1074/jbc.M414187200},\n journal = {Journal of Biological Chemistry},\n number = {13}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Two dimers of the replication terminator protein (RTP) of Bacillus subtilis bind to a chromosomal DNA terminator site to effect polar replication fork arrest. Cooperative binding of the dimers to overlapping half-sites within the terminator is essential for arrest. It was suggested previously that polarity of fork arrest is the result of the RTP dimer at the blocking (proximal) side within the complex binding very tightly and the permissive-side RTP dimer binding relatively weakly. In order to investigate this \"differential binding affinity\" model, we have constructed a series of mutant terminators that contain half-sites of widely different RTP binding affinities in various combinations. Although there appeared to be a correlation between binding affinity at the proximal half-site and fork arrest efficiency in vivo for some terminators, several deviated significantly from this correlation. Some terminators exhibited greatly reduced binding cooperativity (and therefore have reduced affinity at each half-site) but were highly efficient in fork arrest, whereas one terminator had normal affinity over the proximal half-site, yet had low fork arrest efficiency. The results show clearly that there is no direct correlation between the RTP binding affinity (either within the full complex or at the proximal half-site within the full complex) and the efficiency of replication fork arrest in vivo. Thus, the differential binding affinity over the proximal and distal half-sites cannot be solely responsible for functional polarity of fork arrest. Furthermore, efficient fork arrest relies on features in addition to the tight binding of RTP to terminator DNA. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"pursglove-mackay-cslanotchabovetherest-2005\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  CSL: A notch above the rest.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Pursglove,  S.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>International Journal of Biochemistry and Cell Biology</i>, 37(12): 2472-2477.  2005.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.biocel.2005.06.013\"\n     onclick=\"log_download('pursglove-mackay-cslanotchabovetherest-2005', 'http://doi.org/10.1016/j.biocel.2005.06.013', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/pursglove-mackay-cslanotchabovetherest-2005\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('pursglove-mackay-cslanotchabovetherest-2005')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {CSL: A notch above the rest},\n type = {article},\n year = {2005},\n pages = {2472-2477},\n volume = {37},\n id = {951f9e11-35e5-393f-985a-6df0a62be9ff},\n created = {2023-01-10T01:46:05.458Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:05.458Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {CSL (CBF1, Suppressor of Hairless, Lag-1) is a transcription factor that is responsible for activating the genes downstream of the Notch signalling pathway, a pathway that is essential for the development of the nervous system and the differentiation of the haematopoietic system among others. In the absence of Notch signalling, CSL represses transcription of Notch target genes, and following activation by Notch, CSL is converted into a transcriptional activator and activates transcription of the same genes. These two opposing functions of CSL are mediated through interactions with distinct protein complexes. The Notch signalling pathway and its crucial cofactor CSL can maintain cells in an undifferentiated state, and have therefore been associated with a growing list of cancers. In addition, CSL has been co-opted by Epstein-Barr virus to mediate viral and host gene transcription following infection. © 2005 Elsevier Ltd. All rights reserved.},\n bibtype = {article},\n author = {Pursglove, S.E. and Mackay, J.P.},\n doi = {10.1016/j.biocel.2005.06.013},\n journal = {International Journal of Biochemistry and Cell Biology},\n number = {12}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  CSL (CBF1, Suppressor of Hairless, Lag-1) is a transcription factor that is responsible for activating the genes downstream of the Notch signalling pathway, a pathway that is essential for the development of the nervous system and the differentiation of the haematopoietic system among others. In the absence of Notch signalling, CSL represses transcription of Notch target genes, and following activation by Notch, CSL is converted into a transcriptional activator and activates transcription of the same genes. These two opposing functions of CSL are mediated through interactions with distinct protein complexes. The Notch signalling pathway and its crucial cofactor CSL can maintain cells in an undifferentiated state, and have therefore been associated with a growing list of cancers. In addition, CSL has been co-opted by Epstein-Barr virus to mediate viral and host gene transcription following infection. © 2005 Elsevier Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2004\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2004')\"\n      onkeypress=\"toggleGroup('group_2004')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2004</span>\n      <span class=\"bibbase_group_count\">\n        \n        (9)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfrom1hnmrcirculardichroismandpredictionstudies-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/15099568\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfrom1hnmrcirculardichroismandpredictionstudies-2004', 'http://www.ncbi.nlm.nih.gov/pubmed/15099568', event);\">\n    Presence of transient helical segments in the galanin-like peptide evident from 1H NMR, circular dichroism, and prediction studies.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dastmalchi,  S.; Church,  W.; Morris,  M., B.; Iismaa,  T., P.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Structural Biology</i>, 146(3): 261-271. 6 2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/15099568\"\n     onclick=\"log_download('dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfrom1hnmrcirculardichroismandpredictionstudies-2004', 'http://www.ncbi.nlm.nih.gov/pubmed/15099568', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Presence of transient helical segments in the galanin-like peptide evident from 1H NMR, circular dichroism, and prediction studies [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jsb.2004.01.004\"\n     onclick=\"log_download('dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfrom1hnmrcirculardichroismandpredictionstudies-2004', 'http://doi.org/10.1016/j.jsb.2004.01.004', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfrom1hnmrcirculardichroismandpredictionstudies-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfrom1hnmrcirculardichroismandpredictionstudies-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Presence of transient helical segments in the galanin-like peptide evident from 1H NMR, circular dichroism, and prediction studies},\n type = {article},\n year = {2004},\n keywords = {Circular dichroism,Galanin-like peptide,NMR spectroscopy,Nascent helix,Secondary structure prediction},\n pages = {261-271},\n volume = {146},\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/15099568},\n month = {6},\n id = {ccd45448-027a-3ba1-94ee-88d2de266d92},\n created = {2020-12-17T05:29:55.417Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:55.417Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Dastmalchi2004},\n source_type = {ARTICLE},\n notes = {cited By 7},\n private_publication = {false},\n abstract = {Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods.},\n bibtype = {article},\n author = {Dastmalchi, Siavoush and Church, W.Bret and Morris, Michael B. and Iismaa, Tiina P. and Mackay, Joel P.},\n doi = {10.1016/j.jsb.2004.01.004},\n journal = {Journal of Structural Biology},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"feng-gell-zhou-gu-kong-li-hu-yan-etal-molecularmechanismofahspmediatedstabilizationofhemoglobin-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Molecular mechanism of AHSP-mediated stabilization of α-hemoglobin.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Feng,  L.; Gell,  D.; Zhou,  S.; Gu,  L.; Kong,  Y.; Li,  J.; Hu,  M.; Yan,  N.; Lee,  C.; Rich,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Shi,  Y.\n</span>\n\n  \n\n</span>\n\n  <i>Cell</i>, 119(5): 629-640.  2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.cell.2004.11.025\"\n     onclick=\"log_download('feng-gell-zhou-gu-kong-li-hu-yan-etal-molecularmechanismofahspmediatedstabilizationofhemoglobin-2004', 'http://doi.org/10.1016/j.cell.2004.11.025', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/feng-gell-zhou-gu-kong-li-hu-yan-etal-molecularmechanismofahspmediatedstabilizationofhemoglobin-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('feng-gell-zhou-gu-kong-li-hu-yan-etal-molecularmechanismofahspmediatedstabilizationofhemoglobin-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Molecular mechanism of AHSP-mediated stabilization of α-hemoglobin},\n type = {article},\n year = {2004},\n pages = {629-640},\n volume = {119},\n id = {ed2d19cb-f91e-3151-b6da-974c5bb50868},\n created = {2023-01-10T01:46:06.370Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:06.370Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Hemoglobin A (HbA), the oxygen delivery system in humans, comprises two α and two β subunits. Free α-hemoglobin (αHb) is unstable, and its precipitation contributes to the pathophysiology of β thalassemia. In erythrocytes, the α-hemoglobin stabilizing protein (AHSP) binds αHb and inhibits its precipitation. The crystal structure of AHSP bound to Fe(II)-αHb reveals that AHSP specifically recognizes the G and H helices of αHb through a hydrophobic interface that largely recapitulates the α1-β1 interface of hemoglobin. The AHSP-αHb interactions are extensive but suboptimal, explaining why β-hemoglobin can competitively displace AHSP to form HbA. Remarkably, the Fe(II)-heme group in AHSP bound αHb is coordinated by the distal but not the proximal histidine. Importantly, binding to AHSP facilitates the conversion of oxy-αHb to a deoxygenated, oxidized [Fe(III)], nonreactive form in which all six coordinate positions are occupied. These observations reveal the molecular mechanisms by which AHSP stabilizes free αHb.},\n bibtype = {article},\n author = {Feng, L. and Gell, D.A. and Zhou, S. and Gu, L. and Kong, Y. and Li, J. and Hu, M. and Yan, N. and Lee, C. and Rich, A.M. and MacKay, J.P. and Shi, Y.},\n doi = {10.1016/j.cell.2004.11.025},\n journal = {Cell},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Hemoglobin A (HbA), the oxygen delivery system in humans, comprises two α and two β subunits. Free α-hemoglobin (αHb) is unstable, and its precipitation contributes to the pathophysiology of β thalassemia. In erythrocytes, the α-hemoglobin stabilizing protein (AHSP) binds αHb and inhibits its precipitation. The crystal structure of AHSP bound to Fe(II)-αHb reveals that AHSP specifically recognizes the G and H helices of αHb through a hydrophobic interface that largely recapitulates the α1-β1 interface of hemoglobin. The AHSP-αHb interactions are extensive but suboptimal, explaining why β-hemoglobin can competitively displace AHSP to form HbA. Remarkably, the Fe(II)-heme group in AHSP bound αHb is coordinated by the distal but not the proximal histidine. Importantly, binding to AHSP facilitates the conversion of oxy-αHb to a deoxygenated, oxidized [Fe(III)], nonreactive form in which all six coordinate positions are occupied. These observations reveal the molecular mechanisms by which AHSP stabilizes free αHb.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"simpson-lee-bartle-sum-visvader-matthews-mackay-crossley-classiczincfingerfromfriendofgatamediatesaninteractionwiththecoiledcoiloftransformingacidiccoiledcoil3-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Classic zinc finger from friend of GATA mediates an interaction with the coiled-coil of transforming acidic coiled-coil 3.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Simpson,  R.; Lee,  S.; Bartle,  N.; Sum,  E.; Visvader,  J.; Matthews,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 279(38): 39789-39797.  2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M404130200\"\n     onclick=\"log_download('simpson-lee-bartle-sum-visvader-matthews-mackay-crossley-classiczincfingerfromfriendofgatamediatesaninteractionwiththecoiledcoiloftransformingacidiccoiledcoil3-2004', 'http://doi.org/10.1074/jbc.M404130200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/simpson-lee-bartle-sum-visvader-matthews-mackay-crossley-classiczincfingerfromfriendofgatamediatesaninteractionwiththecoiledcoiloftransformingacidiccoiledcoil3-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('simpson-lee-bartle-sum-visvader-matthews-mackay-crossley-classiczincfingerfromfriendofgatamediatesaninteractionwiththecoiledcoiloftransformingacidiccoiledcoil3-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Classic zinc finger from friend of GATA mediates an interaction with the coiled-coil of transforming acidic coiled-coil 3},\n type = {article},\n year = {2004},\n pages = {39789-39797},\n volume = {279},\n id = {8ae82118-628a-3d0f-85fb-40ef4ede1b72},\n created = {2023-01-10T01:46:07.273Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:07.273Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Classic zinc finger domains (cZFs) consist of a β-hairpin followed by an α-helix. They are among the most abundant of all protein domains and are often found in tandem arrays in DNA-binding proteins, with each finger contributing an α-helix to effect sequence-specific DNA recognition. Lone cZFs, not found in tandem arrays, have been postulated to function in protein interactions. We have studied the transcriptional co-regulator Friend of GATA (FOG), which contains nine zinc fingers. We have discovered that the third cZF of FOG contacts a coiled-coil domain in the centrosomal protein transforming acidic coiled-coil 3 (TACC3). Although FOG-ZF3 exhibited low solubility, we have used a combination of mutational mapping and protein engineering to generate a derivative that was suitable for in vitro and structural analysis. We report that the α-helix of FOG-ZF3 recognizes a C-terminal portion of the TACC3 coiled-coil. Remarkably, the α-helical surface utilized by FOG-ZF3 is the same surface responsible for the well established sequence-specific DNA-binding properties of many other cZFs. Our data demonstrate the versatility of cZFs and have implications for the analysis of many as yet uncharacterized cZF proteins.},\n bibtype = {article},\n author = {Simpson, R.J.Y. and Lee, S.H.Y. and Bartle, N. and Sum, E.Y. and Visvader, J.E. and Matthews, J.M. and Mackay, J.P. and Crossley, M.},\n doi = {10.1074/jbc.M404130200},\n journal = {Journal of Biological Chemistry},\n number = {38}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Classic zinc finger domains (cZFs) consist of a β-hairpin followed by an α-helix. They are among the most abundant of all protein domains and are often found in tandem arrays in DNA-binding proteins, with each finger contributing an α-helix to effect sequence-specific DNA recognition. Lone cZFs, not found in tandem arrays, have been postulated to function in protein interactions. We have studied the transcriptional co-regulator Friend of GATA (FOG), which contains nine zinc fingers. We have discovered that the third cZF of FOG contacts a coiled-coil domain in the centrosomal protein transforming acidic coiled-coil 3 (TACC3). Although FOG-ZF3 exhibited low solubility, we have used a combination of mutational mapping and protein engineering to generate a derivative that was suitable for in vitro and structural analysis. We report that the α-helix of FOG-ZF3 recognizes a C-terminal portion of the TACC3 coiled-coil. Remarkably, the α-helical surface utilized by FOG-ZF3 is the same surface responsible for the well established sequence-specific DNA-binding properties of many other cZFs. Our data demonstrate the versatility of cZFs and have implications for the analysis of many as yet uncharacterized cZF proteins.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfromsup1suphnmrcirculardichroismandpredictionstudies-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Presence of transient helical segments in the galanin-like peptide evident from <sup>1</sup>H NMR, circular dichroism, and prediction studies.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dastmalchi,  S.; Church,  W.; Morris,  M.; Iismaa,  T.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Structural Biology</i>, 146(3): 261-271.  2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.jsb.2004.01.004\"\n     onclick=\"log_download('dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfromsup1suphnmrcirculardichroismandpredictionstudies-2004', 'http://doi.org/10.1016/j.jsb.2004.01.004', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfromsup1suphnmrcirculardichroismandpredictionstudies-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dastmalchi-church-morris-iismaa-mackay-presenceoftransienthelicalsegmentsinthegalaninlikepeptideevidentfromsup1suphnmrcirculardichroismandpredictionstudies-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Presence of transient helical segments in the galanin-like peptide evident from &lt;sup&gt;1&lt;/sup&gt;H NMR, circular dichroism, and prediction studies},\n type = {article},\n year = {2004},\n pages = {261-271},\n volume = {146},\n id = {dcdf6b5e-2e76-3021-abce-66a56e5b7f57},\n created = {2023-01-10T01:46:08.193Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:08.193Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods. © 2004 Elsevier Inc. All rights reserved.},\n bibtype = {article},\n author = {Dastmalchi, S. and Church, W.B. and Morris, M.B. and Iismaa, T.P. and Mackay, J.P.},\n doi = {10.1016/j.jsb.2004.01.004},\n journal = {Journal of Structural Biology},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP encompasses within its sequence the first 13 residues of galanin, known to be crucial to binding and activation of galanin receptor (GalR) subtypes. Using 2D-NMR and circular dichroism spectroscopy we demonstrated that GALP does not adopt a preferred conformation in pure water alone. However, it shows characteristics of transient turn-like structures in two distinct regions of its sequence, 11-23 and 41-49. These transient ordered structures, nascent helices, probably form stable helical structures upon addition of the helix-inducing solvent, trifluoroethanol, as determined by circular dichroism studies. Secondary structure prediction methods also predict the presence of two helical regions in the sequence of GALP overlapping reasonably with those regions identified as nascent helical structures by experimental methods. © 2004 Elsevier Inc. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"chow-mackay-whisstock-scanlon-bottomley-structuralandfunctionalanalysisofthejosephindomainofthepolyglutamineproteinataxin3-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural and functional analysis of the Josephin domain of the polyglutamine protein ataxin-3.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Chow,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>; Whisstock,  J.; Scanlon,  M.;  and Bottomley,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemical and Biophysical Research Communications</i>, 322(2): 387-394.  2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.bbrc.2004.07.131\"\n     onclick=\"log_download('chow-mackay-whisstock-scanlon-bottomley-structuralandfunctionalanalysisofthejosephindomainofthepolyglutamineproteinataxin3-2004', 'http://doi.org/10.1016/j.bbrc.2004.07.131', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/chow-mackay-whisstock-scanlon-bottomley-structuralandfunctionalanalysisofthejosephindomainofthepolyglutamineproteinataxin3-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('chow-mackay-whisstock-scanlon-bottomley-structuralandfunctionalanalysisofthejosephindomainofthepolyglutamineproteinataxin3-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural and functional analysis of the Josephin domain of the polyglutamine protein ataxin-3},\n type = {article},\n year = {2004},\n pages = {387-394},\n volume = {322},\n id = {74e20e29-48a8-34da-ac85-4beac421f531},\n created = {2023-01-10T01:46:09.118Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:09.118Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Ataxin-3 belongs to the family of polyglutamine proteins, which are associated with nine different neurodegenerative disorders. Relatively little is known about the structural and functional properties of ataxin-3, and only recently have these aspects of the protein begun to be explored. We have performed a preliminary investigation into the conserved N-terminal domain of ataxin-3, termed Josephin. We show that Josephin is a monomeric domain which folds into a globular conformation and possesses ubiquitin protease activity. In addition, we demonstrate that the presence of the polyglutamine region of the protein does not alter the structure of the protein. However, its presence destabilizes the Josephin domain. The implications of these data in the pathogenesis of polyglutamine repeat proteins are discussed. © 2004 Elsevier Inc. All rights reserved.},\n bibtype = {article},\n author = {Chow, M.K.M. and MacKay, J.P. and Whisstock, J.C. and Scanlon, M.J. and Bottomley, S.P.},\n doi = {10.1016/j.bbrc.2004.07.131},\n journal = {Biochemical and Biophysical Research Communications},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Ataxin-3 belongs to the family of polyglutamine proteins, which are associated with nine different neurodegenerative disorders. Relatively little is known about the structural and functional properties of ataxin-3, and only recently have these aspects of the protein begun to be explored. We have performed a preliminary investigation into the conserved N-terminal domain of ataxin-3, termed Josephin. We show that Josephin is a monomeric domain which folds into a globular conformation and possesses ubiquitin protease activity. In addition, we demonstrate that the presence of the polyglutamine region of the protein does not alter the structure of the protein. However, its presence destabilizes the Josephin domain. The implications of these data in the pathogenesis of polyglutamine repeat proteins are discussed. © 2004 Elsevier Inc. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"donadini-liew-kwan-mackay-fields-crystalandsolutionstructuresofasuperantigenfromyersiniapseudotuberculosisrevealayellyrollfold-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Crystal and Solution Structures of a Superantigen from Yersinia pseudotuberculosis Reveal a Yelly-Roll Fold.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Donadini,  R.; Liew,  C.; Kwan,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Fields,  B.\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 12(1): 145-156.  2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/j.str.2003.12.002\"\n     onclick=\"log_download('donadini-liew-kwan-mackay-fields-crystalandsolutionstructuresofasuperantigenfromyersiniapseudotuberculosisrevealayellyrollfold-2004', 'http://doi.org/10.1016/j.str.2003.12.002', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/donadini-liew-kwan-mackay-fields-crystalandsolutionstructuresofasuperantigenfromyersiniapseudotuberculosisrevealayellyrollfold-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('donadini-liew-kwan-mackay-fields-crystalandsolutionstructuresofasuperantigenfromyersiniapseudotuberculosisrevealayellyrollfold-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Crystal and Solution Structures of a Superantigen from Yersinia pseudotuberculosis Reveal a Yelly-Roll Fold},\n type = {article},\n year = {2004},\n pages = {145-156},\n volume = {12},\n id = {cff79b19-ac8f-3b35-9aa3-73d032e0181d},\n created = {2023-01-10T01:46:10.033Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:10.033Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Superantigens are a class of microbial proteins with the ability to excessively activate T cells by binding to the T cell receptor. The staphylococcal and streptococcal superantigens are closely related in structure and possess an N-terminal domain that resembles an OB fold and a C-terminal domain similar to a β-grasp fold. Yersinia pseudotuberculosis produces superantigens, YPMa, YPMb, and YPMc, which have no significant amino acid similarity to other proteins. We have determined the crystal and solution structures of YPMa, which show that the protein has a jelly-roll fold. The closest structural neighbors to YPMa are viral capsid proteins and members of the tumor necrosis factor superfamily. In the crystal structure, YPMa packs as a trimer, another feature shared with viral capsid proteins and TNF superfamily proteins. However, in solution YPMa behaves as a monomer, and any functional relevance of the trimer observed in the crystals is yet to be established.},\n bibtype = {article},\n author = {Donadini, R. and Liew, C.W. and Kwan, A.H.Y. and Mackay, J.P. and Fields, B.A.},\n doi = {10.1016/j.str.2003.12.002},\n journal = {Structure},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Superantigens are a class of microbial proteins with the ability to excessively activate T cells by binding to the T cell receptor. The staphylococcal and streptococcal superantigens are closely related in structure and possess an N-terminal domain that resembles an OB fold and a C-terminal domain similar to a β-grasp fold. Yersinia pseudotuberculosis produces superantigens, YPMa, YPMb, and YPMc, which have no significant amino acid similarity to other proteins. We have determined the crystal and solution structures of YPMa, which show that the protein has a jelly-roll fold. The closest structural neighbors to YPMa are viral capsid proteins and members of the tumor necrosis factor superfamily. In the crystal structure, YPMa packs as a trimer, another feature shared with viral capsid proteins and TNF superfamily proteins. However, in solution YPMa behaves as a monomer, and any functional relevance of the trimer observed in the crystals is yet to be established.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kong-zhou-kihm-katein-yu-gell-mackay-adachi-etal-lossofhemoglobinstabilizingproteinimpairserythropoiesisandexacerbatesthalassemia-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Loss of α-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates β-thalassemia.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kong,  Y.; Zhou,  S.; Kihm,  A.; Katein,  A.; Yu,  X.; Gell,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Adachi,  K.; Foster-Brown,  L.; Louden,  C.; Gow,  A.;  and Weiss,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Clinical Investigation</i>, 114(10): 1457-1466.  2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1172/JCI21982\"\n     onclick=\"log_download('kong-zhou-kihm-katein-yu-gell-mackay-adachi-etal-lossofhemoglobinstabilizingproteinimpairserythropoiesisandexacerbatesthalassemia-2004', 'http://doi.org/10.1172/JCI21982', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kong-zhou-kihm-katein-yu-gell-mackay-adachi-etal-lossofhemoglobinstabilizingproteinimpairserythropoiesisandexacerbatesthalassemia-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kong-zhou-kihm-katein-yu-gell-mackay-adachi-etal-lossofhemoglobinstabilizingproteinimpairserythropoiesisandexacerbatesthalassemia-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Loss of α-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates β-thalassemia},\n type = {article},\n year = {2004},\n pages = {1457-1466},\n volume = {114},\n id = {b699fcb7-add4-3ff6-a946-b7bf6448eaeb},\n created = {2023-01-10T01:46:10.944Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:10.944Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Hemoglobin (Hb) A production during red blood cell development is coordinated to minimize the deleterious effects of free α- and β-Hb subunits, which are unstable and cytotoxic. The α-Hb-stabilizing protein (AHSP) is an erythroid protein that specifically binds α-Hb and prevents its precipitation in vitro, which suggests that it may function to limit free α-Hb toxicities in vivo. We investigated this possibility through gene ablation and biochemical studies. AHSP-/- erythrocytes contained hemoglobin precipitates and were short-lived. In hematopoietic tissues, erythroid precursors were elevated in number but exhibited increased apoptosis. Consistent with unstable α-Hb, AHSP-/- erythrocytes contained increased ROS and evidence of oxidative damage. Moreover, purified recombinant AHSP inhibited ROS production by α-Hb in solution. Finally, loss of AHSP worsened the phenotype of β-thalassemia, a common inherited anemia characterized by excess free α-Hb. Together, the data support a model in which AHSP binds α-Hb transiently to stabilize its conformation and render it biochemically inert prior to Hb A assembly. This function is essential for normal erythropoiesis and, to a greater extent, in β-thalassemia. Our findings raise the possibility that altered AHSP expression levels could modulate the severity of β-thalassemia in humans.},\n bibtype = {article},\n author = {Kong, Y. and Zhou, S. and Kihm, A.J. and Katein, A.M. and Yu, X. and Gell, D.A. and Mackay, J.P. and Adachi, K. and Foster-Brown, L. and Louden, C.S. and Gow, A.J. and Weiss, M.J.},\n doi = {10.1172/JCI21982},\n journal = {Journal of Clinical Investigation},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Hemoglobin (Hb) A production during red blood cell development is coordinated to minimize the deleterious effects of free α- and β-Hb subunits, which are unstable and cytotoxic. The α-Hb-stabilizing protein (AHSP) is an erythroid protein that specifically binds α-Hb and prevents its precipitation in vitro, which suggests that it may function to limit free α-Hb toxicities in vivo. We investigated this possibility through gene ablation and biochemical studies. AHSP-/- erythrocytes contained hemoglobin precipitates and were short-lived. In hematopoietic tissues, erythroid precursors were elevated in number but exhibited increased apoptosis. Consistent with unstable α-Hb, AHSP-/- erythrocytes contained increased ROS and evidence of oxidative damage. Moreover, purified recombinant AHSP inhibited ROS production by α-Hb in solution. Finally, loss of AHSP worsened the phenotype of β-thalassemia, a common inherited anemia characterized by excess free α-Hb. Together, the data support a model in which AHSP binds α-Hb transiently to stabilize its conformation and render it biochemically inert prior to Hb A assembly. This function is essential for normal erythropoiesis and, to a greater extent, in β-thalassemia. Our findings raise the possibility that altered AHSP expression levels could modulate the severity of β-thalassemia in humans.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"westman-perdomo-matthews-crossley-mackay-structuralstudiesonaproteinbindingzincfingerdomainofeosrevealbothsimilaritiesanddifferencestoclassicalzincfingers-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural studies on a protein-binding zinc-finger domain of eos reveal both similarities and differences to classical zinc fingers.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Westman,  B.; Perdomo,  J.; Matthews,  J.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 43(42): 13318-13327.  2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi049506a\"\n     onclick=\"log_download('westman-perdomo-matthews-crossley-mackay-structuralstudiesonaproteinbindingzincfingerdomainofeosrevealbothsimilaritiesanddifferencestoclassicalzincfingers-2004', 'http://doi.org/10.1021/bi049506a', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/westman-perdomo-matthews-crossley-mackay-structuralstudiesonaproteinbindingzincfingerdomainofeosrevealbothsimilaritiesanddifferencestoclassicalzincfingers-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('westman-perdomo-matthews-crossley-mackay-structuralstudiesonaproteinbindingzincfingerdomainofeosrevealbothsimilaritiesanddifferencestoclassicalzincfingers-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural studies on a protein-binding zinc-finger domain of eos reveal both similarities and differences to classical zinc fingers},\n type = {article},\n year = {2004},\n pages = {13318-13327},\n volume = {43},\n id = {08b79ba5-e49f-31ea-9efb-6f22acd29fae},\n created = {2023-01-10T01:46:11.867Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:11.867Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The oligomerization domain that is present at the C terminus of Ikaros-family proteins and the protein Trps-1 is important for the proper regulation of developmental processes such as hematopoiesis. Remarkably, this domain is predicted to contain two classical zinc fingers (ZnFs), domains normally associated with the recognition of nucleic acids. The preference for protein binding by these predicted ZnFs is not well-understood. We have used a range of methods to gain insight into the structure of this domain. Circular dichroism, UV - vis, and NMR experiments carried out on the C-terminal domain of Eos (EosC) revealed that the two putative ZnFs (C1 and C2) are separable, i.e., capable of folding independently in the presence of ZnII. We next determined the structure of EosC2 using NMR spectroscopy, revealing that, although the overall fold of EosC2 is similar to other classical ZnFs, a number of differences exist. For example, the conformation of the C terminus of EosC2 appears to be flexible and may result in a major rearrangement of the zinc ligands. Finally, alanine-scanning mutagenesis was used to identify the residues that are involved in the homo- and hetero-oligomerization of Eos, and these results are discussed in the context of the structure of EosC. These studies provide the first structural insights into how EosC mediates protein - protein interactions and contributes to our understanding of why it does not exhibit high-affinity DNA binding.},\n bibtype = {article},\n author = {Westman, B.J. and Perdomo, J. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},\n doi = {10.1021/bi049506a},\n journal = {Biochemistry},\n number = {42}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The oligomerization domain that is present at the C terminus of Ikaros-family proteins and the protein Trps-1 is important for the proper regulation of developmental processes such as hematopoiesis. Remarkably, this domain is predicted to contain two classical zinc fingers (ZnFs), domains normally associated with the recognition of nucleic acids. The preference for protein binding by these predicted ZnFs is not well-understood. We have used a range of methods to gain insight into the structure of this domain. Circular dichroism, UV - vis, and NMR experiments carried out on the C-terminal domain of Eos (EosC) revealed that the two putative ZnFs (C1 and C2) are separable, i.e., capable of folding independently in the presence of ZnII. We next determined the structure of EosC2 using NMR spectroscopy, revealing that, although the overall fold of EosC2 is similar to other classical ZnFs, a number of differences exist. For example, the conformation of the C terminus of EosC2 appears to be flexible and may result in a major rearrangement of the zinc ligands. Finally, alanine-scanning mutagenesis was used to identify the residues that are involved in the homo- and hetero-oligomerization of Eos, and these results are discussed in the context of the structure of EosC. These studies provide the first structural insights into how EosC mediates protein - protein interactions and contributes to our understanding of why it does not exhibit high-affinity DNA binding.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"sunde-mcgrath-young-matthews-chua-mackay-death-tc1isanoveltumorigenicandnativelydisorderedproteinassociatedwiththyroidcancer-2004\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  TC-1 Is a Novel Tumorigenic and Natively Disordered Protein Associated with Thyroid Cancer.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Sunde,  M.; McGrath,  K.; Young,  L.; Matthews,  J.; Chua,  E.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Death,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Cancer Research</i>, 64(8): 2766-2773.  2004.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1158/0008-5472.CAN-03-2093\"\n     onclick=\"log_download('sunde-mcgrath-young-matthews-chua-mackay-death-tc1isanoveltumorigenicandnativelydisorderedproteinassociatedwiththyroidcancer-2004', 'http://doi.org/10.1158/0008-5472.CAN-03-2093', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/sunde-mcgrath-young-matthews-chua-mackay-death-tc1isanoveltumorigenicandnativelydisorderedproteinassociatedwiththyroidcancer-2004\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('sunde-mcgrath-young-matthews-chua-mackay-death-tc1isanoveltumorigenicandnativelydisorderedproteinassociatedwiththyroidcancer-2004')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {TC-1 Is a Novel Tumorigenic and Natively Disordered Protein Associated with Thyroid Cancer},\n type = {article},\n year = {2004},\n pages = {2766-2773},\n volume = {64},\n id = {17d8f871-48a9-351a-8a2f-85f939c4f286},\n created = {2023-01-10T01:46:12.786Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:12.786Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {A novel gene, thyroid cancer 1 (TC-1), was found recently to be overexpressed in thyroid cancer. TC-1 shows no homology to any of the known thyroid cancer-associated genes. We have produced stable transformants of normal thyroid cells that express the TC-1 gene, and these cells show increased proliferation rates and anchorage-independent growth in soft agar. Apoptosis rates also are decreased in the transformed cells. We also have expressed recombinant TC-1 protein and have undertaken a structural and functional characterization of the protein. The protein is monomeric and predominantly unstructured under conditions of physiologic salt and pH. This places it in the category of natively disordered proteins, a rapidly expanding group of proteins, many members of which play critical roles in cell regulation processes. We show that the protein can be phosphorylated by cyclic AMP-dependent protein kinase and protein kinase C, and the activity of both of these kinases is up-regulated when cells are stably transfected with TC-1. These results suggest that overexpression of TC-1 may be important in thyroid carcinogenesis.},\n bibtype = {article},\n author = {Sunde, M. and McGrath, K.C.Y. and Young, L. and Matthews, J.M. and Chua, E.L. and Mackay, J.P. and Death, A.K.},\n doi = {10.1158/0008-5472.CAN-03-2093},\n journal = {Cancer Research},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A novel gene, thyroid cancer 1 (TC-1), was found recently to be overexpressed in thyroid cancer. TC-1 shows no homology to any of the known thyroid cancer-associated genes. We have produced stable transformants of normal thyroid cells that express the TC-1 gene, and these cells show increased proliferation rates and anchorage-independent growth in soft agar. Apoptosis rates also are decreased in the transformed cells. We also have expressed recombinant TC-1 protein and have undertaken a structural and functional characterization of the protein. The protein is monomeric and predominantly unstructured under conditions of physiologic salt and pH. This places it in the category of natively disordered proteins, a rapidly expanding group of proteins, many members of which play critical roles in cell regulation processes. We show that the protein can be phosphorylated by cyclic AMP-dependent protein kinase and protein kinase C, and the activity of both of these kinases is up-regulated when cells are stably transfected with TC-1. These results suggest that overexpression of TC-1 may be important in thyroid carcinogenesis.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2003\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2003')\"\n      onkeypress=\"toggleGroup('group_2003')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2003</span>\n      <span class=\"bibbase_group_count\">\n        \n        (8)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"westman-perdomo-sunde-crossley-mackay-thecterminaldomainofeosformsahighordercomplexinsolution-2003\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The C-terminal Domain of Eos Forms a High Order Complex in Solution.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Westman,  B.; Perdomo,  J.; Sunde,  M.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 278(43): 42419-42426.  2003.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M306817200\"\n     onclick=\"log_download('westman-perdomo-sunde-crossley-mackay-thecterminaldomainofeosformsahighordercomplexinsolution-2003', 'http://doi.org/10.1074/jbc.M306817200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/westman-perdomo-sunde-crossley-mackay-thecterminaldomainofeosformsahighordercomplexinsolution-2003\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('westman-perdomo-sunde-crossley-mackay-thecterminaldomainofeosformsahighordercomplexinsolution-2003')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The C-terminal Domain of Eos Forms a High Order Complex in Solution},\n type = {article},\n year = {2003},\n pages = {42419-42426},\n volume = {278},\n id = {d44e19e1-2ac4-30ab-88a3-a1966922ed83},\n created = {2023-01-10T01:46:13.722Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:13.722Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Ikaros family transcription factors play important roles in the control of hematopoiesis. Family members are predicted to contain up to six classic zinc fingers that are arranged into N- and C-terminal domains. The N-terminal domain is responsible for site-specific DNA binding, whereas the C-terminal domain primarily mediates the homo- and hetero-oligomerization between family members. Although the mechanisms of action of these proteins are not completely understood, the zinc finger domains are known to play a central role. In the current study, we have sought to understand the physical and functional properties of these domains, in particular the C-terminal domain. We show that the N-terminal domain from Eos, and not its C-terminal region, is required to recognize GGGA consensus sequences. Surprisingly, in contrast to the behavior exhibited by Ikaros, the C-terminal domain of Eos inhibits the DNA-binding activity of the full-length protein. In addition, we have used a range of biophysical techniques to demonstrate that the C-terminal domain of Eos mediates the formation of complexes that consist of nine or ten molecules. These results constitute the first direct demonstration that Ikaros family proteins can form higher order complexes in solution, and we discuss this unexpected result in the context of what is currently known about the family members and their possible mechanism of action.},\n bibtype = {article},\n author = {Westman, B.J. and Perdomo, J. and Sunde, M. and Crossley, M. and Mackay, J.P.},\n doi = {10.1074/jbc.M306817200},\n journal = {Journal of Biological Chemistry},\n number = {43}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Ikaros family transcription factors play important roles in the control of hematopoiesis. Family members are predicted to contain up to six classic zinc fingers that are arranged into N- and C-terminal domains. The N-terminal domain is responsible for site-specific DNA binding, whereas the C-terminal domain primarily mediates the homo- and hetero-oligomerization between family members. Although the mechanisms of action of these proteins are not completely understood, the zinc finger domains are known to play a central role. In the current study, we have sought to understand the physical and functional properties of these domains, in particular the C-terminal domain. We show that the N-terminal domain from Eos, and not its C-terminal region, is required to recognize GGGA consensus sequences. Surprisingly, in contrast to the behavior exhibited by Ikaros, the C-terminal domain of Eos inhibits the DNA-binding activity of the full-length protein. In addition, we have used a range of biophysical techniques to demonstrate that the C-terminal domain of Eos mediates the formation of complexes that consist of nine or ten molecules. These results constitute the first direct demonstration that Ikaros family proteins can form higher order complexes in solution, and we discuss this unexpected result in the context of what is currently known about the family members and their possible mechanism of action.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kwan-czolij-mackay-crossley-pentaprobeacomprehensivesequencefortheonestepdetectionofdnabindingactivities-2003\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Pentaprobe: a comprehensive sequence for the one-step detection of DNA-binding activities.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kwan,  A.; Czolij,  R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Nucleic acids research</i>, 31(20).  2003.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1093/nar/gng124\"\n     onclick=\"log_download('kwan-czolij-mackay-crossley-pentaprobeacomprehensivesequencefortheonestepdetectionofdnabindingactivities-2003', 'http://doi.org/10.1093/nar/gng124', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kwan-czolij-mackay-crossley-pentaprobeacomprehensivesequencefortheonestepdetectionofdnabindingactivities-2003\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kwan-czolij-mackay-crossley-pentaprobeacomprehensivesequencefortheonestepdetectionofdnabindingactivities-2003')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Pentaprobe: a comprehensive sequence for the one-step detection of DNA-binding activities.},\n type = {article},\n year = {2003},\n volume = {31},\n id = {82c033ec-4b64-301f-b28b-440885f5251a},\n created = {2023-01-10T01:46:14.623Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:14.623Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The rapid increase in the number of novel proteins identified in genome projects necessitates simple and rapid methods for assigning function. We describe a strategy for determining whether novel proteins possess typical sequence-specific DNA-binding activity. Many proteins bind recognition sequences of 5 bp or less. Given that there are 4(5) possible 5 bp sites, one might expect the length of sequence required to cover all possibilities would be 4(5) x 5 or 5120 nt. But by allowing overlaps, utilising both strands and using a computer algorithm to generate the minimum sequence, we find the length required is only 516 base pairs. We generated this sequence as six overlapping double-stranded oligonucleotides, termed pentaprobe, and used it in gel retardation experiments to assess DNA binding by both known and putative DNA-binding proteins from several protein families. We have confirmed binding by the zinc finger proteins BKLF, Eos and Pegasus, the Ets domain protein PU.1 and the treble clef N- and C-terminal fingers of GATA-1. We also showed that the N-terminal zinc finger domain of FOG-1 does not behave as a typical DNA-binding domain. Our results suggest that pentaprobe, and related sequences such as hexaprobe, represent useful tools for probing protein function.},\n bibtype = {article},\n author = {Kwan, A.H. and Czolij, R. and Mackay, J.P. and Crossley, M.},\n doi = {10.1093/nar/gng124},\n journal = {Nucleic acids research},\n number = {20}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The rapid increase in the number of novel proteins identified in genome projects necessitates simple and rapid methods for assigning function. We describe a strategy for determining whether novel proteins possess typical sequence-specific DNA-binding activity. Many proteins bind recognition sequences of 5 bp or less. Given that there are 4(5) possible 5 bp sites, one might expect the length of sequence required to cover all possibilities would be 4(5) x 5 or 5120 nt. But by allowing overlaps, utilising both strands and using a computer algorithm to generate the minimum sequence, we find the length required is only 516 base pairs. We generated this sequence as six overlapping double-stranded oligonucleotides, termed pentaprobe, and used it in gel retardation experiments to assess DNA binding by both known and putative DNA-binding proteins from several protein families. We have confirmed binding by the zinc finger proteins BKLF, Eos and Pegasus, the Ets domain protein PU.1 and the treble clef N- and C-terminal fingers of GATA-1. We also showed that the N-terminal zinc finger domain of FOG-1 does not behave as a typical DNA-binding domain. Our results suggest that pentaprobe, and related sequences such as hexaprobe, represent useful tools for probing protein function.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"plambeck-kwan-adams-westman-vanderweyden-medcalf-morris-mackay-thestructureofthezincfingerdomainfromhumansplicingfactorznf265fold-2003\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The structure of the zinc finger domain from human splicing factor ZNF265 fold.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Plambeck,  C.; Kwan,  A.; Adams,  D.; Westman,  B.; Van der Weyden,  L.; Medcalf,  R.; Morris,  B.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 278(25): 22805-22811.  2003.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M301896200\"\n     onclick=\"log_download('plambeck-kwan-adams-westman-vanderweyden-medcalf-morris-mackay-thestructureofthezincfingerdomainfromhumansplicingfactorznf265fold-2003', 'http://doi.org/10.1074/jbc.M301896200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/plambeck-kwan-adams-westman-vanderweyden-medcalf-morris-mackay-thestructureofthezincfingerdomainfromhumansplicingfactorznf265fold-2003\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('plambeck-kwan-adams-westman-vanderweyden-medcalf-morris-mackay-thestructureofthezincfingerdomainfromhumansplicingfactorznf265fold-2003')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The structure of the zinc finger domain from human splicing factor ZNF265 fold},\n type = {article},\n year = {2003},\n pages = {22805-22811},\n volume = {278},\n id = {762d5635-24ff-39d3-85a7-d0f832df931b},\n created = {2023-01-10T01:46:15.539Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:15.539Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Identification of the protein domains that are responsible for RNA recognition has lagged behind the characterization of protein-DNA interactions. However, it is now becoming clear that a range of structural motifs bind to RNA and their structures and molecular mechanisms of action are beginning to be elucidated. In this report, we have expressed and purified one of the two putative RNA-binding domains from ZNF265, a protein that has been shown to bind to the spliceosomal components U1-70K and U2AF35 and to direct alternative splicing. We show that this domain, which contains four highly conserved cysteine residues, forms a stable, monomeric structure upon the addition of 1 molar eq of Zn(II). Determination of the solution structure of this domain reveals a conformation comprising two stacked β-hairpins oriented at ·80° to each other and sandwiching the zinc ion; the fold resembles the zinc ribbon class of zinc-binding domains, although with one less β-strand than most members of the class. Analysis of the structure reveals a striking resemblance to known RNA-binding motifs in terms of the distribution of key surface residues responsible for making RNA contacts, despite a complete lack of structural homology. Furthermore, we have used an RNA gel shift assay to demonstrate that a single crossed finger domain from ZNF265 is capable of binding to an RNA message. Taken together, these results define a new RNA-binding motif and should provide insight into the functions of the &gt;100 uncharacterized proteins in the sequence data bases that contain this domain.},\n bibtype = {article},\n author = {Plambeck, C.A. and Kwan, A.H.Y. and Adams, D.J. and Westman, B.J. and Van der Weyden, L. and Medcalf, R.L. and Morris, B.J. and Mackay, J.P.},\n doi = {10.1074/jbc.M301896200},\n journal = {Journal of Biological Chemistry},\n number = {25}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Identification of the protein domains that are responsible for RNA recognition has lagged behind the characterization of protein-DNA interactions. However, it is now becoming clear that a range of structural motifs bind to RNA and their structures and molecular mechanisms of action are beginning to be elucidated. In this report, we have expressed and purified one of the two putative RNA-binding domains from ZNF265, a protein that has been shown to bind to the spliceosomal components U1-70K and U2AF35 and to direct alternative splicing. We show that this domain, which contains four highly conserved cysteine residues, forms a stable, monomeric structure upon the addition of 1 molar eq of Zn(II). Determination of the solution structure of this domain reveals a conformation comprising two stacked β-hairpins oriented at ·80° to each other and sandwiching the zinc ion; the fold resembles the zinc ribbon class of zinc-binding domains, although with one less β-strand than most members of the class. Analysis of the structure reveals a striking resemblance to known RNA-binding motifs in terms of the distribution of key surface residues responsible for making RNA contacts, despite a complete lack of structural homology. Furthermore, we have used an RNA gel shift assay to demonstrate that a single crossed finger domain from ZNF265 is capable of binding to an RNA message. Taken together, these results define a new RNA-binding motif and should provide insight into the functions of the >100 uncharacterized proteins in the sequence data bases that contain this domain.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"deane-mackay-kwan-sum-visvader-matthews-structuralbasisfortherecognitionofidb1bythenterminallimdomainsoflmo2andlmo4-2003\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Structural basis for the recognition of Idb1 by the N-terminal LIM domains of LMO2 and LMO4.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Deane,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Kwan,  A.; Sum,  E.; Visvader,  J.;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>EMBO Journal</i>, 22(9): 2224-2233.  2003.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1093/emboj/cdg196\"\n     onclick=\"log_download('deane-mackay-kwan-sum-visvader-matthews-structuralbasisfortherecognitionofidb1bythenterminallimdomainsoflmo2andlmo4-2003', 'http://doi.org/10.1093/emboj/cdg196', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/deane-mackay-kwan-sum-visvader-matthews-structuralbasisfortherecognitionofidb1bythenterminallimdomainsoflmo2andlmo4-2003\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('deane-mackay-kwan-sum-visvader-matthews-structuralbasisfortherecognitionofidb1bythenterminallimdomainsoflmo2andlmo4-2003')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Structural basis for the recognition of Idb1 by the N-terminal LIM domains of LMO2 and LMO4},\n type = {article},\n year = {2003},\n pages = {2224-2233},\n volume = {22},\n id = {443069ef-b4a5-37f0-958b-48eb15aacddd},\n created = {2023-01-10T01:46:16.462Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:16.462Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {LMO2 and LMO4 are members of a small family of nuclear transcriptional regulators that are important for both normal development and disease processes. LMO2 is essential for hemopoiesis and angiogenesis, and inappropriate overexpression of this protein leads to T-cell leukemias. LMO4 is developmentally regulated in the mammary gland and has been implicated in breast oncogenesis. Both proteins comprise two tandemly repeated LIM domains. LMO2 and LMO4 interact with the ubiquitous nuclear adaptor protein Idb1/NLI/CLIM2, which associates with the LIM domains of LMO and LIM homeodomain proteins via its LIM interaction domain (Idb1-LID). We report the solution structures of two LMO:Idb1 complexes (PDB: 1M3V and 1J20) and show that Idb1-LID binds to the N-terminal LIM domain (LIM1) of LMO2 and LMO4 in an extended conformation, contributing a third strand to a β-hairpin in LIM1 domains. These findings constitute the first molecular definition of LIM-mediated protein-protein interactions and suggest a mechanism by which Idb1 can bind a variety of LIM domains that share low sequence homology.},\n bibtype = {article},\n author = {Deane, J.E. and Mackay, J.P. and Kwan, A.H.Y. and Sum, E.Y.M. and Visvader, J.E. and Matthews, J.M.},\n doi = {10.1093/emboj/cdg196},\n journal = {EMBO Journal},\n number = {9}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  LMO2 and LMO4 are members of a small family of nuclear transcriptional regulators that are important for both normal development and disease processes. LMO2 is essential for hemopoiesis and angiogenesis, and inappropriate overexpression of this protein leads to T-cell leukemias. LMO4 is developmentally regulated in the mammary gland and has been implicated in breast oncogenesis. Both proteins comprise two tandemly repeated LIM domains. LMO2 and LMO4 interact with the ubiquitous nuclear adaptor protein Idb1/NLI/CLIM2, which associates with the LIM domains of LMO and LIM homeodomain proteins via its LIM interaction domain (Idb1-LID). We report the solution structures of two LMO:Idb1 complexes (PDB: 1M3V and 1J20) and show that Idb1-LID binds to the N-terminal LIM domain (LIM1) of LMO2 and LMO4 in an extended conformation, contributing a third strand to a β-hairpin in LIM1 domains. These findings constitute the first molecular definition of LIM-mediated protein-protein interactions and suggest a mechanism by which Idb1 can bind a variety of LIM domains that share low sequence homology.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kwan-gell-verger-crossley-matthews-mackay-engineeringaproteinscaffoldfromaphdfinger-2003\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Engineering a protein scaffold from a PHD finger.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Gell,  D.; Verger,  A.; Crossley,  M.; Matthews,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 11(7): 803-813.  2003.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0969-2126(03)00122-9\"\n     onclick=\"log_download('kwan-gell-verger-crossley-matthews-mackay-engineeringaproteinscaffoldfromaphdfinger-2003', 'http://doi.org/10.1016/S0969-2126(03)00122-9', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kwan-gell-verger-crossley-matthews-mackay-engineeringaproteinscaffoldfromaphdfinger-2003\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kwan-gell-verger-crossley-matthews-mackay-engineeringaproteinscaffoldfromaphdfinger-2003')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Engineering a protein scaffold from a PHD finger},\n type = {article},\n year = {2003},\n pages = {803-813},\n volume = {11},\n id = {b8887e04-25ff-3420-9eae-4d046cdc7ab2},\n created = {2023-01-10T01:46:17.361Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:17.361Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The design of proteins with tailored functions remains a relatively elusive goal. Small size, a well-defined structure, and the ability to maintain structural integrity despite multiple mutations are all desirable properties for such designer proteins. Many zinc binding domains fit this description. We determined the structure of a PHD finger from the transcriptional cofactor Mi2β and investigated the suitability of this domain as a scaffold for presenting selected binding functions. The two flexible loops in the structure were mutated extensively by either substitution or expansion, without affecting the overall fold of the domain. A binding site for the corepressor CtBP2 was also grafted onto the domain, creating a new PHD domain that can specifically bind CtBP2 both in vitro and in the context of a eukaryotic cell nucleus. These results represent a step toward designing new regulatory proteins for modulating aberrant gene expression in vivo.},\n bibtype = {article},\n author = {Kwan, A.H.Y. and Gell, D.A. and Verger, A. and Crossley, M. and Matthews, J.M. and Mackay, J.P.},\n doi = {10.1016/S0969-2126(03)00122-9},\n journal = {Structure},\n number = {7}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The design of proteins with tailored functions remains a relatively elusive goal. Small size, a well-defined structure, and the ability to maintain structural integrity despite multiple mutations are all desirable properties for such designer proteins. Many zinc binding domains fit this description. We determined the structure of a PHD finger from the transcriptional cofactor Mi2β and investigated the suitability of this domain as a scaffold for presenting selected binding functions. The two flexible loops in the structure were mutated extensively by either substitution or expansion, without affecting the overall fold of the domain. A binding site for the corepressor CtBP2 was also grafted onto the domain, creating a new PHD domain that can specifically bind CtBP2 both in vitro and in the context of a eukaryotic cell nucleus. These results represent a step toward designing new regulatory proteins for modulating aberrant gene expression in vivo.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kook-lepore-gitler-lu-yung-mackay-zhou-ferrari-etal-cardiachypertrophyandhistonedeacetylasedependenttranscriptionalrepressionmediatedbytheatypicalhomeodomainproteinhop-2003\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Cardiac hypertrophy and histone deacetylase-dependent transcriptional repression mediated by the atypical homeodomain protein Hop.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kook,  H.; Lepore,  J.; Gitler,  A.; Lu,  M.; Yung,  W.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Zhou,  R.; Ferrari,  V.; Gruber,  P.;  and Epstein,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Clinical Investigation</i>, 112(6): 863-871.  2003.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1172/JCI19137\"\n     onclick=\"log_download('kook-lepore-gitler-lu-yung-mackay-zhou-ferrari-etal-cardiachypertrophyandhistonedeacetylasedependenttranscriptionalrepressionmediatedbytheatypicalhomeodomainproteinhop-2003', 'http://doi.org/10.1172/JCI19137', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kook-lepore-gitler-lu-yung-mackay-zhou-ferrari-etal-cardiachypertrophyandhistonedeacetylasedependenttranscriptionalrepressionmediatedbytheatypicalhomeodomainproteinhop-2003\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kook-lepore-gitler-lu-yung-mackay-zhou-ferrari-etal-cardiachypertrophyandhistonedeacetylasedependenttranscriptionalrepressionmediatedbytheatypicalhomeodomainproteinhop-2003')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Cardiac hypertrophy and histone deacetylase-dependent transcriptional repression mediated by the atypical homeodomain protein Hop},\n type = {article},\n year = {2003},\n pages = {863-871},\n volume = {112},\n id = {ea3de67e-e52c-3b2d-8ffd-ffe82aeeec88},\n created = {2023-01-10T01:46:18.276Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:18.276Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Activation of multiple pathways is associated with cardiac hypertrophy and heart failure. Repression of antihypertrophic pathways has rarely been demonstrated to cause cardiac hypertrophy in vivo. Hop is an unusual homeodomain protein that is expressed by embryonic and postnatal cardiac myocytes. Unlike other homeodomain proteins, Hop does not bind DNA. Rather, it modulates cardiac growth and proliferation by inhibiting the transcriptional activity of serum response factor (SRF) in cardiomyocytes. Here we show that Hop can inhibit SRF-dependent transcriptional activation by recruiting histone deacetylase (HDAC) activity and can form a complex that includes HDAC2. Transgenic mice that overexpress Hop develop severe cardiac hypertrophy, cardiac fibrosis, and premature death. A mutant form of Hop, which does not recruit HDAC activity, does not induce hypertrophy. Treatment of Hop transgenic mice with trichostatin A, an HDAC inhibitor, prevents hypertrophy. In addition trichostatin A also attenuates hypertrophy induced by infusion of isoproterenol. Thus, chromatin remodeling and repression of otherwise active transcriptional processes can result in hypertrophy and heart failure, and this process can be blocked with chemical HDAC inhibitors.},\n bibtype = {article},\n author = {Kook, H. and Lepore, J.J. and Gitler, A.D. and Lu, M.M. and Yung, W.W.-M. and Mackay, J. and Zhou, R. and Ferrari, V. and Gruber, P. and Epstein, J.A.},\n doi = {10.1172/JCI19137},\n journal = {Journal of Clinical Investigation},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Activation of multiple pathways is associated with cardiac hypertrophy and heart failure. Repression of antihypertrophic pathways has rarely been demonstrated to cause cardiac hypertrophy in vivo. Hop is an unusual homeodomain protein that is expressed by embryonic and postnatal cardiac myocytes. Unlike other homeodomain proteins, Hop does not bind DNA. Rather, it modulates cardiac growth and proliferation by inhibiting the transcriptional activity of serum response factor (SRF) in cardiomyocytes. Here we show that Hop can inhibit SRF-dependent transcriptional activation by recruiting histone deacetylase (HDAC) activity and can form a complex that includes HDAC2. Transgenic mice that overexpress Hop develop severe cardiac hypertrophy, cardiac fibrosis, and premature death. A mutant form of Hop, which does not recruit HDAC activity, does not induce hypertrophy. Treatment of Hop transgenic mice with trichostatin A, an HDAC inhibitor, prevents hypertrophy. In addition trichostatin A also attenuates hypertrophy induced by infusion of isoproterenol. Thus, chromatin remodeling and repression of otherwise active transcriptional processes can result in hypertrophy and heart failure, and this process can be blocked with chemical HDAC inhibitors.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"simpson-cram-czolij-matthews-crossley-mackay-cchxzincfingerderivativesretaintheabilitytobindzniiandmediateproteindnainteractions-2003\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  CCHX zinc finger derivatives retain the ability to bind Zn(II) and mediate protein-DNA interactions.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Simpson,  R.; Cram,  E.; Czolij,  R.; Matthews,  J.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 278(30): 28011-28018.  2003.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M211146200\"\n     onclick=\"log_download('simpson-cram-czolij-matthews-crossley-mackay-cchxzincfingerderivativesretaintheabilitytobindzniiandmediateproteindnainteractions-2003', 'http://doi.org/10.1074/jbc.M211146200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/simpson-cram-czolij-matthews-crossley-mackay-cchxzincfingerderivativesretaintheabilitytobindzniiandmediateproteindnainteractions-2003\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('simpson-cram-czolij-matthews-crossley-mackay-cchxzincfingerderivativesretaintheabilitytobindzniiandmediateproteindnainteractions-2003')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {CCHX zinc finger derivatives retain the ability to bind Zn(II) and mediate protein-DNA interactions},\n type = {article},\n year = {2003},\n pages = {28011-28018},\n volume = {278},\n id = {254bb9d7-8c93-373a-9c53-0d7edd74d55b},\n created = {2023-01-10T01:46:19.216Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:19.216Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Classical (CCHH) zinc fingers are among the most common protein domains found in eukaryotes. They function as molecular recognition elements that mediate specific contact with DNA, RNA, or other proteins and are composed of a ββα fold surrounding a single zinc ion that is ligated by two cysteine and two histidine residues. In a number of variant zinc fingers, the final histidine is not conserved, and in other unrelated zinc binding domains, residues such as aspartate can function as zinc ligands. To test whether the final histidine is required for normal folding and the DNA-binding function of classical zinc fingers, we focused on finger 3 of basic Krüppel-like factor. The structure of this domain was determined using NMR spectroscopy and found to constitute a typical classical zinc finger. We generated a panel of substitution mutants at the final histidine in this finger and found that several of the mutants retained some ability to fold in the presence of zinc. Consistent with this result, we showed that mutation of the final histidine had only a modest effect on DNA binding in the context of the full three-finger DNA-binding domain of basic Krüppel-like factor. Further, the zinc binding ability of one of the point mutants was tested and found to be indistinguishable from the wild-type domain. These results suggest that the final zinc chelating histidine is not an essential feature of classical zinc fingers and have implications for zinc finger evolution, regulation, and the design of experiments testing the functional roles of these domains.},\n bibtype = {article},\n author = {Simpson, R.J.Y. and Cram, E.D. and Czolij, R. and Matthews, J.M. and Crossley, M. and Mackay, J.P.},\n doi = {10.1074/jbc.M211146200},\n journal = {Journal of Biological Chemistry},\n number = {30}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Classical (CCHH) zinc fingers are among the most common protein domains found in eukaryotes. They function as molecular recognition elements that mediate specific contact with DNA, RNA, or other proteins and are composed of a ββα fold surrounding a single zinc ion that is ligated by two cysteine and two histidine residues. In a number of variant zinc fingers, the final histidine is not conserved, and in other unrelated zinc binding domains, residues such as aspartate can function as zinc ligands. To test whether the final histidine is required for normal folding and the DNA-binding function of classical zinc fingers, we focused on finger 3 of basic Krüppel-like factor. The structure of this domain was determined using NMR spectroscopy and found to constitute a typical classical zinc finger. We generated a panel of substitution mutants at the final histidine in this finger and found that several of the mutants retained some ability to fold in the presence of zinc. Consistent with this result, we showed that mutation of the final histidine had only a modest effect on DNA binding in the context of the full three-finger DNA-binding domain of basic Krüppel-like factor. Further, the zinc binding ability of one of the point mutants was tested and found to be indistinguishable from the wild-type domain. These results suggest that the final zinc chelating histidine is not an essential feature of classical zinc fingers and have implications for zinc finger evolution, regulation, and the design of experiments testing the functional roles of these domains.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"proudfoot-mackay-karuso-solutionstructureandbehaviourofmrurrpicchxnmeinf2infphisup2supbynmrspectroscopyandmolecularmodelling-2003\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Solution structure and behaviour of Δ-m-α-[Ru(R,R- picchxnMe<inf>2</inf>)(phi)]<sup>2+</sup> by NMR spectroscopy and molecular modelling.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Proudfoot,  E.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Karuso,  P.\n</span>\n\n  \n\n</span>\n\n  <i>Dalton Transactions</i>, (2): 165-170.  2003.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1039/b208846k\"\n     onclick=\"log_download('proudfoot-mackay-karuso-solutionstructureandbehaviourofmrurrpicchxnmeinf2infphisup2supbynmrspectroscopyandmolecularmodelling-2003', 'http://doi.org/10.1039/b208846k', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/proudfoot-mackay-karuso-solutionstructureandbehaviourofmrurrpicchxnmeinf2infphisup2supbynmrspectroscopyandmolecularmodelling-2003\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('proudfoot-mackay-karuso-solutionstructureandbehaviourofmrurrpicchxnmeinf2infphisup2supbynmrspectroscopyandmolecularmodelling-2003')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution structure and behaviour of Δ-m-α-[Ru(R,R- picchxnMe&lt;inf&gt;2&lt;/inf&gt;)(phi)]&lt;sup&gt;2+&lt;/sup&gt; by NMR spectroscopy and molecular modelling},\n type = {article},\n year = {2003},\n pages = {165-170},\n id = {a5866cc0-0e9f-38d7-a4df-ab0bef4ad5e5},\n created = {2023-01-10T01:46:20.125Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:20.125Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The self-association of the DNA metalloprobe Δ-cis-α-[Ru(R,R- picchxnMe2)(phi)]2+ (α-phi) in aqueous solution has been investigated using 1H NMR spectroscopy and molecular modelling. The concentration dependence of proton chemical shifts of the complex gave initial indications of a self-associated species, while its structural isomer Δ-cis-β-[Ru(R,R-picchxnMe2)(phi)]2+ (β-phi) showed no such dependence. 2D-COSY and 2D-ROESY experiments were used for the complete assignment of the proton resonances of both isomers and allowed a qualitative determination of the self-association of the a isomer through the detection of intermolecular ROEs. NMR spectroscopy can also be effectively used to differentiate Δ- and Λ-diastereomers. In addition, we show, by pulsed field gradient longitudinal eddy-current delay (PFGLED) NMR spectroscopy, that α-phi self-associates at higher concentrations with an effective molecular weight at 25 mM three times that at 2.5 mM. This apparent oligomerisation was not observed for the β-isomer. © The Royal Society of Chemistry 2003.},\n bibtype = {article},\n author = {Proudfoot, E.M. and Mackay, J.P. and Karuso, P.},\n doi = {10.1039/b208846k},\n journal = {Dalton Transactions},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The self-association of the DNA metalloprobe Δ-cis-α-[Ru(R,R- picchxnMe2)(phi)]2+ (α-phi) in aqueous solution has been investigated using 1H NMR spectroscopy and molecular modelling. The concentration dependence of proton chemical shifts of the complex gave initial indications of a self-associated species, while its structural isomer Δ-cis-β-[Ru(R,R-picchxnMe2)(phi)]2+ (β-phi) showed no such dependence. 2D-COSY and 2D-ROESY experiments were used for the complete assignment of the proton resonances of both isomers and allowed a qualitative determination of the self-association of the a isomer through the detection of intermolecular ROEs. NMR spectroscopy can also be effectively used to differentiate Δ- and Λ-diastereomers. In addition, we show, by pulsed field gradient longitudinal eddy-current delay (PFGLED) NMR spectroscopy, that α-phi self-associates at higher concentrations with an effective molecular weight at 25 mM three times that at 2.5 mM. This apparent oligomerisation was not observed for the β-isomer. © The Royal Society of Chemistry 2003.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2002\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2002')\"\n      onkeypress=\"toggleGroup('group_2002')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2002</span>\n      <span class=\"bibbase_group_count\">\n        \n        (13)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/12127581\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002', 'http://www.ncbi.nlm.nih.gov/pubmed/12127581', event);\">\n    Ikaros: a key regulator of haematopoiesis.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Westman,  B., J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Gell,  D.\n</span>\n\n  \n\n</span>\n\n  <i>The International Journal of Biochemistry & Cell Biology</i>, 34(10): 1304-1307. 10 2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/12127581\"\n     onclick=\"log_download('westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002', 'http://www.ncbi.nlm.nih.gov/pubmed/12127581', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Ikaros: a key regulator of haematopoiesis [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S1357-2725(02)00070-5\"\n     onclick=\"log_download('westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002', 'http://doi.org/10.1016/S1357-2725(02)00070-5', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Ikaros: a key regulator of haematopoiesis},\n type = {article},\n year = {2002},\n keywords = {Haematopoiesis,Ikaros,Pericentromeric-heterochromatin,Transcription},\n pages = {1304-1307},\n volume = {34},\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/12127581},\n month = {10},\n id = {5eef5524-c104-30d5-8fd5-93d7f32bc139},\n created = {2020-12-17T05:29:56.237Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:56.237Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Westman2002},\n source_type = {ARTICLE},\n notes = {cited By 21},\n private_publication = {false},\n abstract = {Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function--unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription.},\n bibtype = {article},\n author = {Westman, Belinda J. and Mackay, Joel P. and Gell, David},\n doi = {10.1016/S1357-2725(02)00070-5},\n journal = {The International Journal of Biochemistry &amp; Cell Biology},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function--unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"dvid-yi-eatona-weissc-mackaye-biophysicalcharacterizationoftheglobinbindingproteinhemoglobinstabilizingprotein-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Biophysical characterization of the α-globin binding protein α-hemoglobin stabilizing protein.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dvid,  G.; Yi,  K.; Eaton A,  S.; Weiss C,  M.;  and Mackay E,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 277(43): 40602-40609.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M206084200\"\n     onclick=\"log_download('dvid-yi-eatona-weissc-mackaye-biophysicalcharacterizationoftheglobinbindingproteinhemoglobinstabilizingprotein-2002', 'http://doi.org/10.1074/jbc.M206084200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dvid-yi-eatona-weissc-mackaye-biophysicalcharacterizationoftheglobinbindingproteinhemoglobinstabilizingprotein-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dvid-yi-eatona-weissc-mackaye-biophysicalcharacterizationoftheglobinbindingproteinhemoglobinstabilizingprotein-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Biophysical characterization of the α-globin binding protein α-hemoglobin stabilizing protein},\n type = {article},\n year = {2002},\n pages = {40602-40609},\n volume = {277},\n id = {7bd08e1a-428f-320d-b146-14956285c9d5},\n created = {2023-01-10T01:46:21.028Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:21.028Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {α-Hemoglobin stabilizing protein (AHSP) is a small (12 kDa) and abundant erythroid-specific protein that binds specifically to free α-(hemo)globin and prevents its precipitation. When present in excess over β-globin, its normal binding partner, α-globin can have severe cytotoxic effects that contribute to important human diseases such as β-thalassemia. Because AHSP might act as a chaperone to prevent the harmful aggregation of α-globin during normal erythroid cell development and in diseases of globin chain imbalance, it is important to characterize the biochemical properties of the AHSP·α-globin complex. Here we provide the first structural information about AHSP and its interaction with α-globin. We find that AHSP is a predominantly α-helical globular protein with a somewhat asymmetric shape. AHSP and α-globin are both monomeric in solution as determined by analytical ultracentrifugation and bind each other to form a complex with 1:1 subunit stoichiometry, as judged by gel filtration and amino acid analysis. We have used isothermal titration calorimetry to show that the interaction is of moderate affinity with an association constant of 1 × 107 M-1 and is thus likely to be biologically significant given the concentration of AHSP (∼0.1 mM) and hemoglobin (∼4 mm) in the late pro-erythroblast.},\n bibtype = {article},\n author = {Dvid, G. and Yi, K. and Eaton A, S.A. and Weiss C, M.J. and Mackay E, J.P.},\n doi = {10.1074/jbc.M206084200},\n journal = {Journal of Biological Chemistry},\n number = {43}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  α-Hemoglobin stabilizing protein (AHSP) is a small (12 kDa) and abundant erythroid-specific protein that binds specifically to free α-(hemo)globin and prevents its precipitation. When present in excess over β-globin, its normal binding partner, α-globin can have severe cytotoxic effects that contribute to important human diseases such as β-thalassemia. Because AHSP might act as a chaperone to prevent the harmful aggregation of α-globin during normal erythroid cell development and in diseases of globin chain imbalance, it is important to characterize the biochemical properties of the AHSP·α-globin complex. Here we provide the first structural information about AHSP and its interaction with α-globin. We find that AHSP is a predominantly α-helical globular protein with a somewhat asymmetric shape. AHSP and α-globin are both monomeric in solution as determined by analytical ultracentrifugation and bind each other to form a complex with 1:1 subunit stoichiometry, as judged by gel filtration and amino acid analysis. We have used isothermal titration calorimetry to show that the interaction is of moderate affinity with an association constant of 1 × 107 M-1 and is thus likely to be biologically significant given the concentration of AHSP (∼0.1 mM) and hemoglobin (∼4 mm) in the late pro-erythroblast.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"sharpe-matthews-kwan-newton-gell-crossley-mackay-anewzincbindingfoldunderlinestheversatilityofzincbindingmodulesinproteinevolution-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A new zinc binding fold underlines the versatility of zinc binding modules in protein evolution.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Sharpe,  B.; Matthews,  J.; <a class=\"bibbase author link\" href=\"https://www.kwanlabusyd.com/publications-1\">Kwan,  A.</a>; Newton,  A.; Gell,  D.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 10(5): 639-648.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0969-2126(02)00757-8\"\n     onclick=\"log_download('sharpe-matthews-kwan-newton-gell-crossley-mackay-anewzincbindingfoldunderlinestheversatilityofzincbindingmodulesinproteinevolution-2002', 'http://doi.org/10.1016/S0969-2126(02)00757-8', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/sharpe-matthews-kwan-newton-gell-crossley-mackay-anewzincbindingfoldunderlinestheversatilityofzincbindingmodulesinproteinevolution-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('sharpe-matthews-kwan-newton-gell-crossley-mackay-anewzincbindingfoldunderlinestheversatilityofzincbindingmodulesinproteinevolution-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A new zinc binding fold underlines the versatility of zinc binding modules in protein evolution},\n type = {article},\n year = {2002},\n pages = {639-648},\n volume = {10},\n id = {3d10c7ef-8426-3256-9aa7-a918db603a09},\n created = {2023-01-10T01:46:21.952Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:21.952Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Many different zinc binding modules have been identified. Their abundance and variety suggests that the formation of zinc binding folds might be relatively common. We have determined the structure of CH11, a 27-residue peptide derived from the first cysteine/histidine-rich region (CH1) of CREB binding protein (CBP). This peptide forms a highly ordered zinc-dependent fold that is distinct from known folds. The structure differs from a subsequently determined structure of a larger region from the CH3 region of CBP, and the CH11 fold probably represents a nonphysiologically active form. Despite this, the fold is thermostable and tolerant to both multiple alanine mutations and changes in the zinc-ligand spacing. Our data support the idea that zinc binding domains may arise frequently. Additionally, such structures may prove useful as scaffolds for protein design, given their stability and robustness.},\n bibtype = {article},\n author = {Sharpe, B.K. and Matthews, J.M. and Kwan, A.H.Y. and Newton, A. and Gell, D.A. and Crossley, M. and Mackay, J.P.},\n doi = {10.1016/S0969-2126(02)00757-8},\n journal = {Structure},\n number = {5}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Many different zinc binding modules have been identified. Their abundance and variety suggests that the formation of zinc binding folds might be relatively common. We have determined the structure of CH11, a 27-residue peptide derived from the first cysteine/histidine-rich region (CH1) of CREB binding protein (CBP). This peptide forms a highly ordered zinc-dependent fold that is distinct from known folds. The structure differs from a subsequently determined structure of a larger region from the CH3 region of CBP, and the CH11 fold probably represents a nonphysiologically active form. Despite this, the fold is thermostable and tolerant to both multiple alanine mutations and changes in the zinc-ligand spacing. Our data support the idea that zinc binding domains may arise frequently. Additionally, such structures may prove useful as scaffolds for protein design, given their stability and robustness.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"deane-visvader-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofflin4anintramolecularlmo4ldb1complex8-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of FLIN4, an intramolecular LMO4:ldb1 complex [8].\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Deane,  J.; Visvader,  J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biomolecular NMR</i>, 23(2): 165-166.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1023/A:1016363414644\"\n     onclick=\"log_download('deane-visvader-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofflin4anintramolecularlmo4ldb1complex8-2002', 'http://doi.org/10.1023/A:1016363414644', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/deane-visvader-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofflin4anintramolecularlmo4ldb1complex8-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('deane-visvader-mackay-matthews-sup1suphsup15supnandsup13supcassignmentsofflin4anintramolecularlmo4ldb1complex8-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {&lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;15&lt;/sup&gt;N and &lt;sup&gt;13&lt;/sup&gt;C assignments of FLIN4, an intramolecular LMO4:ldb1 complex [8]},\n type = {article},\n year = {2002},\n pages = {165-166},\n volume = {23},\n id = {d2aa00f0-07ce-3979-9e30-49c883eec666},\n created = {2023-01-10T01:46:22.869Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:22.869Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Deane, J.E. and Visvader, J.E. and Mackay, J.P. and Matthews, J.M.},\n doi = {10.1023/A:1016363414644},\n journal = {Journal of Biomolecular NMR},\n number = {2}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"liepinsh-otting-harding-ward-mackay-haymet-solutionstructureofahydrophobicanalogueofthewinterflounderantifreezeprotein-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Solution structure of a hydrophobic analogue of the winter flounder antifreeze protein.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Liepinsh,  E.; Otting,  G.; Harding,  M.; Ward,  L.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Haymet,  A.\n</span>\n\n  \n\n</span>\n\n  <i>European Journal of Biochemistry</i>, 269(4): 1259-1266.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1046/j.1432-1033.2002.02766.x\"\n     onclick=\"log_download('liepinsh-otting-harding-ward-mackay-haymet-solutionstructureofahydrophobicanalogueofthewinterflounderantifreezeprotein-2002', 'http://doi.org/10.1046/j.1432-1033.2002.02766.x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/liepinsh-otting-harding-ward-mackay-haymet-solutionstructureofahydrophobicanalogueofthewinterflounderantifreezeprotein-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('liepinsh-otting-harding-ward-mackay-haymet-solutionstructureofahydrophobicanalogueofthewinterflounderantifreezeprotein-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution structure of a hydrophobic analogue of the winter flounder antifreeze protein},\n type = {article},\n year = {2002},\n pages = {1259-1266},\n volume = {269},\n id = {e7419261-1955-324a-9050-96596e299897},\n created = {2023-01-10T01:46:23.798Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:23.798Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The solution structure of a synthetic mutant type I antifreeze protein (AFP I) was determined in aqueous solution at pH 7.0 using nuclear magnetic resonance (NMR) spectroscopy. The mutations comprised the replacement of the four Thr residues by Val and the introduction of two additional Lys-Glu salt bridges. The antifreeze activity of this mutant peptide, VVVV2KE, has been previously shown to be similar to that of the wild type protein, HPLC6 (defined here as TTTT). The solution structure reveals an ahelix bent in the same direction as the more bent conformer of the published crystalstructure of TTTT, while the side chain χ1 rotamers of VVVV2KE are similar to those of the straighter conformer in the crystal of TTTT. The Val side chains of VVVV2KE assume the same orientations as the Thr side chains of TTTT, confirming the conservative nature of this mutation. The combined data suggest that AFP I undergoes an equilibrium between straight and bent helices in solution, combined with independent equilibria between different side chain rotamers for some of the amino acid residues. The present study presents the first complete sequence-specific resonance assignments and the first complete solution structure determination by NMR of any AFP I protein.},\n bibtype = {article},\n author = {Liepinsh, E. and Otting, G. and Harding, M.M. and Ward, L.G. and Mackay, J.P. and Haymet, A.D.J.},\n doi = {10.1046/j.1432-1033.2002.02766.x},\n journal = {European Journal of Biochemistry},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The solution structure of a synthetic mutant type I antifreeze protein (AFP I) was determined in aqueous solution at pH 7.0 using nuclear magnetic resonance (NMR) spectroscopy. The mutations comprised the replacement of the four Thr residues by Val and the introduction of two additional Lys-Glu salt bridges. The antifreeze activity of this mutant peptide, VVVV2KE, has been previously shown to be similar to that of the wild type protein, HPLC6 (defined here as TTTT). The solution structure reveals an ahelix bent in the same direction as the more bent conformer of the published crystalstructure of TTTT, while the side chain χ1 rotamers of VVVV2KE are similar to those of the straighter conformer in the crystal of TTTT. The Val side chains of VVVV2KE assume the same orientations as the Thr side chains of TTTT, confirming the conservative nature of this mutation. The combined data suggest that AFP I undergoes an equilibrium between straight and bent helices in solution, combined with independent equilibria between different side chain rotamers for some of the amino acid residues. The present study presents the first complete sequence-specific resonance assignments and the first complete solution structure determination by NMR of any AFP I protein.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Ikaros: A key regulator of haematopoiesis.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Westman,  B.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Gell,  D.\n</span>\n\n  \n\n</span>\n\n  <i>International Journal of Biochemistry and Cell Biology</i>, 34(10): 1304-1307.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S1357-2725(02)00070-5\"\n     onclick=\"log_download('westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002', 'http://doi.org/10.1016/S1357-2725(02)00070-5', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('westman-mackay-gell-ikarosakeyregulatorofhaematopoiesis-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Ikaros: A key regulator of haematopoiesis},\n type = {article},\n year = {2002},\n pages = {1304-1307},\n volume = {34},\n id = {b11bff38-f90a-3ab1-9e2f-59aa100f8266},\n created = {2023-01-10T01:46:24.702Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:24.702Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function - unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription. © 2002 Elsevier Science Ltd. All rights reserved.},\n bibtype = {article},\n author = {Westman, B.J. and Mackay, J.P. and Gell, D.},\n doi = {10.1016/S1357-2725(02)00070-5},\n journal = {International Journal of Biochemistry and Cell Biology},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Ikaros is an essential transcription factor for normal lymphocyte development. Because of its interaction with a number of closely related factors, Ikaros is required for correct regulation of differentiation and cell proliferation in T- and B-cell lineages. Interestingly, Ikaros appears to function both as a transcriptional repressor and as an activator through its ability to bind a large number of nuclear factors, including components of both histone deacetylase and ATP-dependent chromatin remodelling complexes. In addition, nuclear localisation is important for Ikaros function - unlike most transcription factors, Ikaros is localised to discrete nuclear foci in lymphoid cells, suggesting it employs novel mechanisms to regulate transcription. © 2002 Elsevier Science Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Characterization of the conserved interaction between GATA and FOG family proteins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kowalski,  K.; Liew,  C.; Matthews,  J.; Gell,  D.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 277(38): 35720-35729.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M204663200\"\n     onclick=\"log_download('kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002', 'http://doi.org/10.1074/jbc.M204663200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Characterization of the conserved interaction between GATA and FOG family proteins},\n type = {article},\n year = {2002},\n pages = {35720-35729},\n volume = {277},\n id = {d4514158-aef2-3645-95c3-a537e76c54ce},\n created = {2023-01-10T01:46:25.606Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:25.606Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.},\n bibtype = {article},\n author = {Kowalski, K. and Liew, C.K. and Matthews, J.M. and Gell, D.A. and Crossley, M. and Mackay, J.P.},\n doi = {10.1074/jbc.M204663200},\n journal = {Journal of Biological Chemistry},\n number = {38}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Characterization of the conserved interaction between GATA and FOG family proteins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kowalski,  K.; Liew,  C.; Matthews,  J.; Gell,  D.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 277(38): 35720-35729.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M204663200\"\n     onclick=\"log_download('kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002', 'http://doi.org/10.1074/jbc.M204663200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kowalski-liew-matthews-gell-crossley-mackay-characterizationoftheconservedinteractionbetweengataandfogfamilyproteins-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Characterization of the conserved interaction between GATA and FOG family proteins},\n type = {article},\n year = {2002},\n pages = {35720-35729},\n volume = {277},\n id = {943dca0a-527a-3b78-8975-163e0bc0a3c9},\n created = {2023-01-10T01:46:26.825Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:26.825Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.},\n bibtype = {article},\n author = {Kowalski, K. and Liew, C.K. and Matthews, J.M. and Gell, D.A. and Crossley, M. and Mackay, J.P.},\n doi = {10.1074/jbc.M204663200},\n journal = {Journal of Biological Chemistry},\n number = {38}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His → Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His → Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"robson-michie-mackay-harry-king-thebacillussubtiliscelldivisionproteinsftslanddivicareintrinsicallyunstableanddonotinteractwithoneanotherintheabsenceofotherseptasomalcomponents-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The Bacillus subtilis cell division proteins FtsL and DivIC are intrinsically unstable and do not interact with one another in the absence of other septasomal components.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Robson,  S.; Michie,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Harry,  E.;  and King,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Molecular Microbiology</i>, 44(3): 663-674.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1046/j.1365-2958.2002.02920.x\"\n     onclick=\"log_download('robson-michie-mackay-harry-king-thebacillussubtiliscelldivisionproteinsftslanddivicareintrinsicallyunstableanddonotinteractwithoneanotherintheabsenceofotherseptasomalcomponents-2002', 'http://doi.org/10.1046/j.1365-2958.2002.02920.x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/robson-michie-mackay-harry-king-thebacillussubtiliscelldivisionproteinsftslanddivicareintrinsicallyunstableanddonotinteractwithoneanotherintheabsenceofotherseptasomalcomponents-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('robson-michie-mackay-harry-king-thebacillussubtiliscelldivisionproteinsftslanddivicareintrinsicallyunstableanddonotinteractwithoneanotherintheabsenceofotherseptasomalcomponents-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The Bacillus subtilis cell division proteins FtsL and DivIC are intrinsically unstable and do not interact with one another in the absence of other septasomal components},\n type = {article},\n year = {2002},\n pages = {663-674},\n volume = {44},\n id = {8a1d4881-9023-34c3-97a4-9fab972fd4c2},\n created = {2023-01-10T01:46:27.729Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:27.729Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The bacterial septum appears to comprise a macromolecular assembly of essential cell division proteins (the &#x27;septasome&#x27;) that are responsible for physically dividing the cell during cytokinesis. FtsL and DivIC are essential components of this division machinery in Bacillus subtilis. We used yeast two-hybrid analysis as well as a variety of biochemical and biophysical methods to examine the proposed interaction between Bacillus subtilis FtsL and DivIC. We show that FtsL and DivIC are thermodynamically unstable proteins that are likely to be unfolded and therefore targeted for degradation unless stabilized by interactions with other components of the septasome. However, we show that this stabilization does not result from a direct interaction between FtsL and DivIC. We propose that the observed interdependence of DivIC and FtsL stability is a result of indirect interactions that are mediated by other septasomal proteins.},\n bibtype = {article},\n author = {Robson, S.A. and Michie, K.A. and Mackay, J.P. and Harry, E. and King, G.F.},\n doi = {10.1046/j.1365-2958.2002.02920.x},\n journal = {Molecular Microbiology},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The bacterial septum appears to comprise a macromolecular assembly of essential cell division proteins (the 'septasome') that are responsible for physically dividing the cell during cytokinesis. FtsL and DivIC are essential components of this division machinery in Bacillus subtilis. We used yeast two-hybrid analysis as well as a variety of biochemical and biophysical methods to examine the proposed interaction between Bacillus subtilis FtsL and DivIC. We show that FtsL and DivIC are thermodynamically unstable proteins that are likely to be unfolded and therefore targeted for degradation unless stabilized by interactions with other components of the septasome. However, we show that this stabilization does not result from a direct interaction between FtsL and DivIC. We propose that the observed interdependence of DivIC and FtsL stability is a result of indirect interactions that are mediated by other septasomal proteins.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"chen-kook-milewski-gitler-lu-li-nazarian-schnepp-etal-hopisanunusualhomeoboxgenethatmodulatescardiacdevelopment-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Hop is an unusual homeobox gene that modulates cardiac development.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Chen,  F.; Kook,  H.; Milewski,  R.; Gitler,  A.; Lu,  M.; Li,  J.; Nazarian,  R.; Schnepp,  R.; Jen,  K.; Biben,  C.; Mullins,  M.;  and Epstein,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Cell</i>, 110(6): 713-723.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0092-8674(02)00932-7\"\n     onclick=\"log_download('chen-kook-milewski-gitler-lu-li-nazarian-schnepp-etal-hopisanunusualhomeoboxgenethatmodulatescardiacdevelopment-2002', 'http://doi.org/10.1016/S0092-8674(02)00932-7', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/chen-kook-milewski-gitler-lu-li-nazarian-schnepp-etal-hopisanunusualhomeoboxgenethatmodulatescardiacdevelopment-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('chen-kook-milewski-gitler-lu-li-nazarian-schnepp-etal-hopisanunusualhomeoboxgenethatmodulatescardiacdevelopment-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Hop is an unusual homeobox gene that modulates cardiac development},\n type = {article},\n year = {2002},\n pages = {713-723},\n volume = {110},\n id = {31c23e13-969a-3713-9fa5-129a498c180a},\n created = {2023-01-10T01:46:28.648Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:28.648Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Hop is a small, divergent homeodomain protein that lacks certain conserved residues required for DNA binding. Hop gene expression initiates early in cardiogenesis and continues in cardiomyocytes throughout embryonic and postnatal development. Genetic and biochemical data indicate that Hop functions directly downstream of Nkx2-5. Inactivation of Hop in mice by homologous recombination results in a partially penetrant embryonic lethal phenotype with severe developmental cardiac defects involving the myocardium. Inhibition of Hop activity in zebrafish embryos likewise disrupts cardiac development and results in severely impaired cardiac function. Hop physically interacts with serum response factor (SRF) and inhibits activation of SRF-dependent transcription by inhibiting SRF binding to DNA. Hop encodes an unusual homeodomain protein that modulates SRF-dependent cardiac-specific gene expression and cardiac development.},\n bibtype = {article},\n author = {Chen, F. and Kook, H. and Milewski, R. and Gitler, A.D. and Lu, M.M. and Li, J. and Nazarian, R. and Schnepp, R. and Jen, K. and Biben, C. and Mullins, M.C. and Epstein, J.A.},\n doi = {10.1016/S0092-8674(02)00932-7},\n journal = {Cell},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Hop is a small, divergent homeodomain protein that lacks certain conserved residues required for DNA binding. Hop gene expression initiates early in cardiogenesis and continues in cardiomyocytes throughout embryonic and postnatal development. Genetic and biochemical data indicate that Hop functions directly downstream of Nkx2-5. Inactivation of Hop in mice by homologous recombination results in a partially penetrant embryonic lethal phenotype with severe developmental cardiac defects involving the myocardium. Inhibition of Hop activity in zebrafish embryos likewise disrupts cardiac development and results in severely impaired cardiac function. Hop physically interacts with serum response factor (SRF) and inhibits activation of SRF-dependent transcription by inhibiting SRF binding to DNA. Hop encodes an unusual homeodomain protein that modulates SRF-dependent cardiac-specific gene expression and cardiac development.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"polekhina-house-traficante-mackay-relaix-sassoon-parker-bowtell-siahubiquitinligaseisstructurallyrelatedtotrafandmodulatestnfsignaling-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Siah ubiquitin ligase is structurally related to traf and modulates tnf-α signaling.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Polekhina,  G.; House,  C.; Traficante,  N.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Relaix,  F.; Sassoon,  D.; Parker,  M.;  and Bowtell,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Nature Structural Biology</i>, 9(1): 68-75.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1038/nsb743\"\n     onclick=\"log_download('polekhina-house-traficante-mackay-relaix-sassoon-parker-bowtell-siahubiquitinligaseisstructurallyrelatedtotrafandmodulatestnfsignaling-2002', 'http://doi.org/10.1038/nsb743', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/polekhina-house-traficante-mackay-relaix-sassoon-parker-bowtell-siahubiquitinligaseisstructurallyrelatedtotrafandmodulatestnfsignaling-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('polekhina-house-traficante-mackay-relaix-sassoon-parker-bowtell-siahubiquitinligaseisstructurallyrelatedtotrafandmodulatestnfsignaling-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Siah ubiquitin ligase is structurally related to traf and modulates tnf-α signaling},\n type = {article},\n year = {2002},\n pages = {68-75},\n volume = {9},\n id = {4c060996-79de-3f5c-ae4a-e1f05aebf808},\n created = {2023-01-10T01:46:29.556Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:29.556Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Members of the Siah (seven in absentia homolog) family of RING domain proteins are components of E3 ubiquitin ligase complexes that catalyze ubiquitination of proteins. We have determined the crystal structure of the substrate-binding domain (SBD) of murine Siah1a to 2.6 Å resolution. The structure reveals that Siah is a dimeric protein and that the SBD adopts an eight-stranded β-sandwich fold that is highly similar to the TRAF-C region of TRAF (TNF-receptor associated factor) proteins. The TRAF-C region interacts with TNF-α receptors and TNF-receptor associated death-domain (TRADD) proteins; however, our findings indicate that these interactions are unlikely to be mimicked by Siah. The Siah structure also reveals two novel zinc fingers in a region with sequence similarity to TRAF. We find that the Siah1a SBD potentiates TNF-α-mediated NF-κB activation. Therefore, Siah proteins share important similarities with the TRAF family of proteins, including their overall domain architecture, three-dimensional structure and functional activity. © 2002 Nature Publishing Group.},\n bibtype = {article},\n author = {Polekhina, G. and House, C.M. and Traficante, N. and Mackay, J.P. and Relaix, F. and Sassoon, D.A. and Parker, M.W. and Bowtell, D.D.L.},\n doi = {10.1038/nsb743},\n journal = {Nature Structural Biology},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Members of the Siah (seven in absentia homolog) family of RING domain proteins are components of E3 ubiquitin ligase complexes that catalyze ubiquitination of proteins. We have determined the crystal structure of the substrate-binding domain (SBD) of murine Siah1a to 2.6 Å resolution. The structure reveals that Siah is a dimeric protein and that the SBD adopts an eight-stranded β-sandwich fold that is highly similar to the TRAF-C region of TRAF (TNF-receptor associated factor) proteins. The TRAF-C region interacts with TNF-α receptors and TNF-receptor associated death-domain (TRADD) proteins; however, our findings indicate that these interactions are unlikely to be mimicked by Siah. The Siah structure also reveals two novel zinc fingers in a region with sequence similarity to TRAF. We find that the Siah1a SBD potentiates TNF-α-mediated NF-κB activation. Therefore, Siah proteins share important similarities with the TRAF family of proteins, including their overall domain architecture, three-dimensional structure and functional activity. © 2002 Nature Publishing Group.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"fairley-westman-pham-haymet-harding-mackay-typeishorthornsculpinantifreezeproteinrecombinantsynthesissolutionconformationandicegrowthinhibitionstudies-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Type I shorthorn sculpin antifreeze protein: Recombinant synthesis, solution conformation, and ice growth inhibition studies.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Fairley,  K.; Westman,  B.; Pham,  L.; Haymet,  A.; Harding,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 277(27): 24073-24080.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M200307200\"\n     onclick=\"log_download('fairley-westman-pham-haymet-harding-mackay-typeishorthornsculpinantifreezeproteinrecombinantsynthesissolutionconformationandicegrowthinhibitionstudies-2002', 'http://doi.org/10.1074/jbc.M200307200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/fairley-westman-pham-haymet-harding-mackay-typeishorthornsculpinantifreezeproteinrecombinantsynthesissolutionconformationandicegrowthinhibitionstudies-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('fairley-westman-pham-haymet-harding-mackay-typeishorthornsculpinantifreezeproteinrecombinantsynthesissolutionconformationandicegrowthinhibitionstudies-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Type I shorthorn sculpin antifreeze protein: Recombinant synthesis, solution conformation, and ice growth inhibition studies},\n type = {article},\n year = {2002},\n pages = {24073-24080},\n volume = {277},\n id = {3b95bafc-8671-34e7-80d0-daa877523c66},\n created = {2023-01-10T01:46:30.477Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:30.477Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {A number of structurally diverse classes of &quot;antifreeze&quot; proteins that allow fish to survive in sub-zero ice-laden waters have been isolated from the blood plasma of cold water teleosts. However, despite receiving a great deal of attention, the one or more mechanisms through which these proteins act are not fully understood. In this report we have synthesized a type I antifreeze polypeptide (AFP) from the shorthorn sculpin Myoxocephalus scorpius using recombinant methods. Construction of a synthetic gene with optimized codon usage and expression as a glutathione S-transferase fusion protein followed by purification yielded milligram amounts of polypeptide with two extra residues appended to the N terminus. Circular dichroism and NMR experiments, including residual dipolar coupling measurements on a 15N-labeled recombinant polypeptide, show that the polypeptides are α-helical with the first four residues being more flexible than the remainder of the sequence. Both the recombinant and synthetic polypeptides modify ice growth, forming facetted crystals just below the freezing point, but display negligible thermal hysteresis. Acetylation of Lys-10, Lys-20, and Lys-21 as well as the N terminus of the recombinant polypeptide gave a derivative that displays both thermal hysteresis (0.4°C at 15 mg/ml) and ice crystal faceting. These results confirm that the N terminus of wild-type polypeptide is functionally important and support our previously proposed mechanism for all type I proteins, in which the hydrophobic face is oriented toward the ice at the ice/water interface.},\n bibtype = {article},\n author = {Fairley, K. and Westman, B.J. and Pham, L.H. and Haymet, A.D.J. and Harding, M.M. and Mackay, J.P.},\n doi = {10.1074/jbc.M200307200},\n journal = {Journal of Biological Chemistry},\n number = {27}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A number of structurally diverse classes of \"antifreeze\" proteins that allow fish to survive in sub-zero ice-laden waters have been isolated from the blood plasma of cold water teleosts. However, despite receiving a great deal of attention, the one or more mechanisms through which these proteins act are not fully understood. In this report we have synthesized a type I antifreeze polypeptide (AFP) from the shorthorn sculpin Myoxocephalus scorpius using recombinant methods. Construction of a synthetic gene with optimized codon usage and expression as a glutathione S-transferase fusion protein followed by purification yielded milligram amounts of polypeptide with two extra residues appended to the N terminus. Circular dichroism and NMR experiments, including residual dipolar coupling measurements on a 15N-labeled recombinant polypeptide, show that the polypeptides are α-helical with the first four residues being more flexible than the remainder of the sequence. Both the recombinant and synthetic polypeptides modify ice growth, forming facetted crystals just below the freezing point, but display negligible thermal hysteresis. Acetylation of Lys-10, Lys-20, and Lys-21 as well as the N terminus of the recombinant polypeptide gave a derivative that displays both thermal hysteresis (0.4°C at 15 mg/ml) and ice crystal faceting. These results confirm that the N terminus of wild-type polypeptide is functionally important and support our previously proposed mechanism for all type I proteins, in which the hydrophobic face is oriented toward the ice at the ice/water interface.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"pursglove-mulhern-mackay-hinds-booker-thesolutionstructureandintramolecularassociationsoftheteckinasesrchomology3domain-2002\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The solution structure and intramolecular associations of the Tec kinase Src homology 3 domain.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Pursglove,  S.; Mulhern,  T.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Hinds,  M.;  and Booker,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 277(1): 755-762.  2002.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M108318200\"\n     onclick=\"log_download('pursglove-mulhern-mackay-hinds-booker-thesolutionstructureandintramolecularassociationsoftheteckinasesrchomology3domain-2002', 'http://doi.org/10.1074/jbc.M108318200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/pursglove-mulhern-mackay-hinds-booker-thesolutionstructureandintramolecularassociationsoftheteckinasesrchomology3domain-2002\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('pursglove-mulhern-mackay-hinds-booker-thesolutionstructureandintramolecularassociationsoftheteckinasesrchomology3domain-2002')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The solution structure and intramolecular associations of the Tec kinase Src homology 3 domain},\n type = {article},\n year = {2002},\n pages = {755-762},\n volume = {277},\n id = {b3f30832-cb31-30ae-a155-a163a5517400},\n created = {2023-01-10T01:46:31.373Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:31.373Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Tec is the prototypic member of a family of intracellular tyrosine kinases that includes Txk, Bmx, Itk, and Btk. Tec family kinases share similarities in domain structure with Src family kinases, but one of the features that differentiates them is a proline-rich region (PRR) preceding their Src homology (SH) 3 domain. Evidence that the PRR of Itk can bind in an intramolecular fashion to its SH3 domain and the lack of a regulatory tyrosine in the C terminus indicates that Tec kinases must be regulated by a different set of intramolecular interactions to the Src kinases. We have determined the solution structure of the Tec SH3 domain and have investigated interactions with its PRR, which contains two SH3-binding sites. We demonstrate that in vitro, the Tec PRR can bind in an intramolecular fashion to the SH3. However, the affinity is lower than that for dimerization via reciprocal PRR-SH3 association. Using site-directed mutagenesis we show that both sites can bind the Tec SH3 domain; site 1 (155KTLPPAP161) binds intramolecularly, while site 2 (165KRRPPPPIPP174) cannot and binds in an intermolecular fashion. These distinct roles for the SH3 binding sites in Tec family kinases could be important for protein targeting and enzyme activation.},\n bibtype = {article},\n author = {Pursglove, S.E. and Mulhern, T.D. and Mackay, J.P. and Hinds, M.G. and Booker, G.W.},\n doi = {10.1074/jbc.M108318200},\n journal = {Journal of Biological Chemistry},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Tec is the prototypic member of a family of intracellular tyrosine kinases that includes Txk, Bmx, Itk, and Btk. Tec family kinases share similarities in domain structure with Src family kinases, but one of the features that differentiates them is a proline-rich region (PRR) preceding their Src homology (SH) 3 domain. Evidence that the PRR of Itk can bind in an intramolecular fashion to its SH3 domain and the lack of a regulatory tyrosine in the C terminus indicates that Tec kinases must be regulated by a different set of intramolecular interactions to the Src kinases. We have determined the solution structure of the Tec SH3 domain and have investigated interactions with its PRR, which contains two SH3-binding sites. We demonstrate that in vitro, the Tec PRR can bind in an intramolecular fashion to the SH3. However, the affinity is lower than that for dimerization via reciprocal PRR-SH3 association. Using site-directed mutagenesis we show that both sites can bind the Tec SH3 domain; site 1 (155KTLPPAP161) binds intramolecularly, while site 2 (165KRRPPPPIPP174) cannot and binds in an intermolecular fashion. These distinct roles for the SH3 binding sites in Tec family kinases could be important for protein targeting and enzyme activation.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2001\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2001')\"\n      onkeypress=\"toggleGroup('group_2001')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2001</span>\n      <span class=\"bibbase_group_count\">\n        \n        (7)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001', 'https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591', event);\">\n    The Hydrophobin EAS Is Largely Unstructured in Solution and Functions by Forming Amyloid-Like Structures.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Matthews,  J., M.; Winefield,  R., D.; Mackay,  L., G.; Haverkamp,  R., G.;  and Templeton,  M., D.\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 9(2): 83-91. 2 2001.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591\"\n     onclick=\"log_download('mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001', 'https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"The Hydrophobin EAS Is Largely Unstructured in Solution and Functions by Forming Amyloid-Like Structures [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0969-2126(00)00559-1\"\n     onclick=\"log_download('mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001', 'http://doi.org/10.1016/S0969-2126(00)00559-1', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The Hydrophobin EAS Is Largely Unstructured in Solution and Functions by Forming Amyloid-Like Structures},\n type = {article},\n year = {2001},\n keywords = {Amphipathic monolayer,EAS,Neurospora crassa,Self-assembly,hydrophobin},\n pages = {83-91},\n volume = {9},\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S0969212600005591},\n month = {2},\n id = {298718cc-0cb5-316f-89d4-dead241698cf},\n created = {2020-12-17T05:29:52.461Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:52.461Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Mackay2001},\n source_type = {ARTICLE},\n notes = {cited By 115},\n private_publication = {false},\n abstract = {Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.},\n bibtype = {article},\n author = {Mackay, Joel P and Matthews, Jacqueline M. and Winefield, Robert D. and Mackay, Lindsey G. and Haverkamp, Richard G. and Templeton, Matthew D.},\n doi = {10.1016/S0969-2126(00)00559-1},\n journal = {Structure},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The hydrophobin EAS is largely unstructured in solution and functions by forming amyloid-like structures.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Matthews,  J.; Winefield,  R.; Mackay,  L.; Haverkamp,  R.;  and Templeton,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 9(2): 83-91.  2001.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0969-2126(00)00559-1\"\n     onclick=\"log_download('mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001', 'http://doi.org/10.1016/S0969-2126(00)00559-1', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-matthews-winefield-mackay-haverkamp-templeton-thehydrophobineasislargelyunstructuredinsolutionandfunctionsbyformingamyloidlikestructures-2001')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The hydrophobin EAS is largely unstructured in solution and functions by forming amyloid-like structures},\n type = {article},\n year = {2001},\n pages = {83-91},\n volume = {9},\n id = {f7eae917-5560-305a-8f43-505a2f44c290},\n created = {2023-01-10T01:46:32.312Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:32.312Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.},\n bibtype = {article},\n author = {Mackay, J.P. and Matthews, J.M. and Winefield, R.D. and Mackay, L.G. and Haverkamp, R.G. and Templeton, M.D.},\n doi = {10.1016/S0969-2126(00)00559-1},\n journal = {Structure},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: Fungal hydrophobin proteins have the remarkable ability to self-assemble into polymeric, amphipathic monolayers on the surface of aerial structures such as spores and fruiting bodies. These monolayers are extremely resistant to degradation and as such offer the possibility of a range of biotechnological applications involving the reversal of surface polarity. The molecular details underlying the formation of these monolayers, however, have been elusive. We have studied EAS, the hydrophobin from the ascomycete Neurospora crassa, in an effort to understand the structural aspects of hydrophobin polymerization. Results: We have purified both wild-type and uniformly 15N-labeled EAS from N. crassa conidia, and used a range of physical methods including multidimensional NMR spectroscopy to provide the first high resolution structural information on a member of the hydrophobin family. We have found that EAS is monomeric but mostly unstructured in solution, except for a small region of antiparallel β sheet that is probably stabilized by four intramolecular disulfide bonds. Polymerised EAS appears to contain substantially higher amounts of β sheet structure, and shares many properties with amyloid fibers, including a characteristic gold-green birefringence under polarized light in the presence of the dye Congo Red. Conclusions: EAS joins an increasing number of proteins that undergo a disorder→order transition in carrying out their normal function. This report is one of the few examples where an amyloid-like state represents the wild-type functional form. Thus the mechanism of amyloid formation, now thought to be a general property of polypeptide chains, has actually been applied in nature to form these remarkable structures.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"wang-connor-wilson-wilson-nicholson-smith-shaw-mackay-etal-discoveryandstructureofapotentandhighlyspecificblockerofinsectcalciumchannels-2001\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Discovery and Structure of a Potent and Highly Specific Blocker of Insect Calcium Channels.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Wang,  X.; Connor,  M.; Wilson,  D.; Wilson,  H.; Nicholson,  G.; Smith,  R.; Shaw,  D.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Alewood,  P.; Christie,  M.; Christie,  M.;  and King,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 276(43): 40306-40312.  2001.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M105206200\"\n     onclick=\"log_download('wang-connor-wilson-wilson-nicholson-smith-shaw-mackay-etal-discoveryandstructureofapotentandhighlyspecificblockerofinsectcalciumchannels-2001', 'http://doi.org/10.1074/jbc.M105206200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/wang-connor-wilson-wilson-nicholson-smith-shaw-mackay-etal-discoveryandstructureofapotentandhighlyspecificblockerofinsectcalciumchannels-2001\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('wang-connor-wilson-wilson-nicholson-smith-shaw-mackay-etal-discoveryandstructureofapotentandhighlyspecificblockerofinsectcalciumchannels-2001')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Discovery and Structure of a Potent and Highly Specific Blocker of Insect Calcium Channels},\n type = {article},\n year = {2001},\n pages = {40306-40312},\n volume = {276},\n id = {6c391a59-29b9-321f-858c-b5248f7b56df},\n created = {2023-01-10T01:46:33.265Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:33.265Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {We have isolated a novel family of insect-selective neurotoxins that appear to be the most potent blockers of insect voltage-gated calcium channels reported to date. These toxins display exceptional phylogenetic specificity, with at least a 10,000-fold preference for insect versus vertebrate calcium channels. The structure of one of the toxins reveals a highly structured, disulfide-rich core and a structurally disordered C-terminal extension that is essential for channel blocking activity. Weak structural/functional homology with ω-agatoxin-IVA/B, the prototypic inhibitor of vertebrate P-type calcium channels, suggests that these two toxin families might share a similar mechanism of action despite their vastly different phylogenetic specificities.},\n bibtype = {article},\n author = {Wang, X.-H. and Connor, M. and Wilson, D. and Wilson, H.I. and Nicholson, G.M. and Smith, R. and Shaw, D. and Mackay, J.P. and Alewood, P.F. and Christie, M.J. and Christie, M.J. and King, G.F.},\n doi = {10.1074/jbc.M105206200},\n journal = {Journal of Biological Chemistry},\n number = {43}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  We have isolated a novel family of insect-selective neurotoxins that appear to be the most potent blockers of insect voltage-gated calcium channels reported to date. These toxins display exceptional phylogenetic specificity, with at least a 10,000-fold preference for insect versus vertebrate calcium channels. The structure of one of the toxins reveals a highly structured, disulfide-rich core and a structurally disordered C-terminal extension that is essential for channel blocking activity. Weak structural/functional homology with ω-agatoxin-IVA/B, the prototypic inhibitor of vertebrate P-type calcium channels, suggests that these two toxin families might share a similar mechanism of action despite their vastly different phylogenetic specificities.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"deane-sum-mackay-lindeman-visvader-matthews-designproductionandcharacterizationofflin2andflin4theengineeringofintramolecularldb1lmocomplexes-2001\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Design, production and characterization of FLIN2 and FLIN4: The engineering of intramolecular ldb1:LMO complexes.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Deane,  J.; Sum,  E.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Lindeman,  G.; Visvader,  J.;  and Matthews,  J.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Engineering</i>, 14(7): 493-499.  2001.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1093/protein/14.7.493\"\n     onclick=\"log_download('deane-sum-mackay-lindeman-visvader-matthews-designproductionandcharacterizationofflin2andflin4theengineeringofintramolecularldb1lmocomplexes-2001', 'http://doi.org/10.1093/protein/14.7.493', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/deane-sum-mackay-lindeman-visvader-matthews-designproductionandcharacterizationofflin2andflin4theengineeringofintramolecularldb1lmocomplexes-2001\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('deane-sum-mackay-lindeman-visvader-matthews-designproductionandcharacterizationofflin2andflin4theengineeringofintramolecularldb1lmocomplexes-2001')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Design, production and characterization of FLIN2 and FLIN4: The engineering of intramolecular ldb1:LMO complexes},\n type = {article},\n year = {2001},\n pages = {493-499},\n volume = {14},\n id = {a9b6f2ec-a058-327e-abad-6aa474786e8a},\n created = {2023-01-10T01:46:34.180Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:34.180Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The nuclear LIM-only (LMO) transcription factors LMO2 and LMO4 play important roles in both normal and leukemic T-cell development. LIM domains are cysteine/histidine-rich domains that contain two structural zinc ions and that function as protein-protein adaptors; members of the LMO family each contain two closely spaced LIM domains. These LMO proteins all bind with high affinity to the nuclear protein LIM domain binding protein 1 (ldb1). The LMO-ldb1 interaction is mediated through the N-terminal LIM domain (LIM1) of LMO proteins and a 38-residue region towards the C-terminus of ldb1 [ldb1(LID)]. Unfortunately, recombinant forms of LMO2 and LMO4 have limited solubility and stability, effectively preventing structural analysis. Therefore, we have designed and constructed a fusion protein in which ldb1(LID) and LIM1 of LMO2 can form an intramolecular complex. The engineered protein, FLIN2 (fusion of the LIM interacting domain of ldb1 and the N-terminal LIM domain of LMO2) has been expressed and purified in milligram quantities. FLIN2 is monomeric, contains significant levels of secondary structure and yields a sharp and well-dispersed one-dimensional 1H NMR spectrum. The analogous LMO4 protein, FLIN4, has almost identical properties. These data suggest that we will be able to obtain high-resolution structural information about the LMO-ldb1 interactions.},\n bibtype = {article},\n author = {Deane, J.E. and Sum, E. and Mackay, J.P. and Lindeman, G.J. and Visvader, J.E. and Matthews, J.M.},\n doi = {10.1093/protein/14.7.493},\n journal = {Protein Engineering},\n number = {7}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The nuclear LIM-only (LMO) transcription factors LMO2 and LMO4 play important roles in both normal and leukemic T-cell development. LIM domains are cysteine/histidine-rich domains that contain two structural zinc ions and that function as protein-protein adaptors; members of the LMO family each contain two closely spaced LIM domains. These LMO proteins all bind with high affinity to the nuclear protein LIM domain binding protein 1 (ldb1). The LMO-ldb1 interaction is mediated through the N-terminal LIM domain (LIM1) of LMO proteins and a 38-residue region towards the C-terminus of ldb1 [ldb1(LID)]. Unfortunately, recombinant forms of LMO2 and LMO4 have limited solubility and stability, effectively preventing structural analysis. Therefore, we have designed and constructed a fusion protein in which ldb1(LID) and LIM1 of LMO2 can form an intramolecular complex. The engineered protein, FLIN2 (fusion of the LIM interacting domain of ldb1 and the N-terminal LIM domain of LMO2) has been expressed and purified in milligram quantities. FLIN2 is monomeric, contains significant levels of secondary structure and yields a sharp and well-dispersed one-dimensional 1H NMR spectrum. The analogous LMO4 protein, FLIN4, has almost identical properties. These data suggest that we will be able to obtain high-resolution structural information about the LMO-ldb1 interactions.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"matthews-visvader-mackay-sup1suphsup15supnandsup13supcassignmentsofflin2anintramolecularlmo2ldb1complex2-2001\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <sup>1</sup>H, <sup>15</sup>N and <sup>13</sup>C assignments of FLIN2, an intramolecular LMO2:ldb1 complex [2].\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Matthews,  J.; Visvader,  J.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biomolecular NMR</i>, 21(4): 385-386.  2001.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1023/A:1013373203772\"\n     onclick=\"log_download('matthews-visvader-mackay-sup1suphsup15supnandsup13supcassignmentsofflin2anintramolecularlmo2ldb1complex2-2001', 'http://doi.org/10.1023/A:1013373203772', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/matthews-visvader-mackay-sup1suphsup15supnandsup13supcassignmentsofflin2anintramolecularlmo2ldb1complex2-2001\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('matthews-visvader-mackay-sup1suphsup15supnandsup13supcassignmentsofflin2anintramolecularlmo2ldb1complex2-2001')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {&lt;sup&gt;1&lt;/sup&gt;H, &lt;sup&gt;15&lt;/sup&gt;N and &lt;sup&gt;13&lt;/sup&gt;C assignments of FLIN2, an intramolecular LMO2:ldb1 complex [2]},\n type = {article},\n year = {2001},\n pages = {385-386},\n volume = {21},\n id = {4480e900-209e-3853-9615-5c2f38cb7107},\n created = {2023-01-10T01:46:35.100Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:35.100Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Matthews, J.M. and Visvader, J.E. and Mackay, J.P.},\n doi = {10.1023/A:1013373203772},\n journal = {Journal of Biomolecular NMR},\n number = {4}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"proudfoot-mackay-karuso-probingsitespecificityofdnabindingmetallointercalatorsbynmrspectroscopyandmolecularmodeling-2001\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Probing site specificity of DNA binding metallointercalators by NMR spectroscopy and molecular modeling.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Proudfoot,  E.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Karuso,  P.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 40(15): 4867-4878.  2001.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi001655f\"\n     onclick=\"log_download('proudfoot-mackay-karuso-probingsitespecificityofdnabindingmetallointercalatorsbynmrspectroscopyandmolecularmodeling-2001', 'http://doi.org/10.1021/bi001655f', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/proudfoot-mackay-karuso-probingsitespecificityofdnabindingmetallointercalatorsbynmrspectroscopyandmolecularmodeling-2001\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('proudfoot-mackay-karuso-probingsitespecificityofdnabindingmetallointercalatorsbynmrspectroscopyandmolecularmodeling-2001')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Probing site specificity of DNA binding metallointercalators by NMR spectroscopy and molecular modeling},\n type = {article},\n year = {2001},\n pages = {4867-4878},\n volume = {40},\n id = {ecd0710f-2f62-38e1-9576-54331818c01f},\n created = {2023-01-10T01:46:35.999Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:35.999Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The molecular recognition of oligonucleotides by chiral ruthenium complexes has been probed by NMR spectroscopy using the template Δ-cis-α- and Δ-cis-β-[Ru(RR-picchxnMe2) (bidentate)]2+, where the bidentate ligand is one of phen (1,10-phenanthroline), dpq (dipyrido[3,2-f:2′,3′-h]quinoxaline), or phi (9,10-phenanthrenequinone diimine) and picchxnMe2 is N,N′-dimethyl-N,N′-di(2-picolyl)-1,2-diaminocyclohexane. By varying only the bidentate ligand in a series of complexes, it was shown that the bidentate alone can alter binding modes. DNA binding studies of the Δ-cis-α,-[Ru(RR-picchxnMe2)(phen)]2+ complex indicate fast exchange kinetics on the chemical shift time scale and a &quot;partial intercalation&quot; mode of binding. This complex binds to [d(CGCGATCGCG)]2 and [d(ATATCGATAT)]2 at AT, TA, and GA sites from the minor groove, as well as to the ends of the oligonucleotide at low temperature. Studies of the Δ-cis-β-[Ru(RR-picchxnMe2)(phen)]2+ complex with [d(CGCGATCGCG)]2 showed that the complex binds only weakly to the ends of the oligonucleotide. The interaction of Δ-cis-α-[Ru(RR-picchxnMe2)-(dpq)]2+ with [d(CGCGATCGCG)]2 showed intermediate exchange kinetics and evidence of minor groove intercalation at the GA base step. In contrast to the phen and dpq complexes, Δ-cis-α- and Δ-cis-β-[Ru-(RR-picchxnMe2)(phi)]2+ showed evidence of major groove binding independent of the metal ion configuration. DNA stabilization induced by complex binding to [d(CGCGATCGCG)]2 (measured as ΔTm) increases in the order phen &lt; dpq and DNA affinity in the order phen &lt; dpq &lt; phi. The groove binding preferences exhibited by the different bidentate ligands is explained with the aid of molecular modeling experiments.},\n bibtype = {article},\n author = {Proudfoot, E.M. and Mackay, J.P. and Karuso, P.},\n doi = {10.1021/bi001655f},\n journal = {Biochemistry},\n number = {15}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The molecular recognition of oligonucleotides by chiral ruthenium complexes has been probed by NMR spectroscopy using the template Δ-cis-α- and Δ-cis-β-[Ru(RR-picchxnMe2) (bidentate)]2+, where the bidentate ligand is one of phen (1,10-phenanthroline), dpq (dipyrido[3,2-f:2′,3′-h]quinoxaline), or phi (9,10-phenanthrenequinone diimine) and picchxnMe2 is N,N′-dimethyl-N,N′-di(2-picolyl)-1,2-diaminocyclohexane. By varying only the bidentate ligand in a series of complexes, it was shown that the bidentate alone can alter binding modes. DNA binding studies of the Δ-cis-α,-[Ru(RR-picchxnMe2)(phen)]2+ complex indicate fast exchange kinetics on the chemical shift time scale and a \"partial intercalation\" mode of binding. This complex binds to [d(CGCGATCGCG)]2 and [d(ATATCGATAT)]2 at AT, TA, and GA sites from the minor groove, as well as to the ends of the oligonucleotide at low temperature. Studies of the Δ-cis-β-[Ru(RR-picchxnMe2)(phen)]2+ complex with [d(CGCGATCGCG)]2 showed that the complex binds only weakly to the ends of the oligonucleotide. The interaction of Δ-cis-α-[Ru(RR-picchxnMe2)-(dpq)]2+ with [d(CGCGATCGCG)]2 showed intermediate exchange kinetics and evidence of minor groove intercalation at the GA base step. In contrast to the phen and dpq complexes, Δ-cis-α- and Δ-cis-β-[Ru-(RR-picchxnMe2)(phi)]2+ showed evidence of major groove binding independent of the metal ion configuration. DNA stabilization induced by complex binding to [d(CGCGATCGCG)]2 (measured as ΔTm) increases in the order phen < dpq and DNA affinity in the order phen < dpq < phi. The groove binding preferences exhibited by the different bidentate ligands is explained with the aid of molecular modeling experiments.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"newton-mackay-crossley-thenterminalzincfingeroftheerythroidtranscriptionfactorgata1bindsgatcmotifsindna-2001\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The N-terminal Zinc Finger of the Erythroid Transcription Factor GATA-1 Binds GATC Motifs in DNA.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Newton,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 276(38): 35794-35801.  2001.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M106256200\"\n     onclick=\"log_download('newton-mackay-crossley-thenterminalzincfingeroftheerythroidtranscriptionfactorgata1bindsgatcmotifsindna-2001', 'http://doi.org/10.1074/jbc.M106256200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/newton-mackay-crossley-thenterminalzincfingeroftheerythroidtranscriptionfactorgata1bindsgatcmotifsindna-2001\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('newton-mackay-crossley-thenterminalzincfingeroftheerythroidtranscriptionfactorgata1bindsgatcmotifsindna-2001')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The N-terminal Zinc Finger of the Erythroid Transcription Factor GATA-1 Binds GATC Motifs in DNA},\n type = {article},\n year = {2001},\n pages = {35794-35801},\n volume = {276},\n id = {668a6be7-8063-3f78-a869-df88d49978b0},\n created = {2023-01-10T01:46:36.902Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:36.902Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The mammalian transcription factor GATA-1 is required for normal erythroid and megakaryocytic development. GATA-1 contains two zinc fingers, the C-terminal finger, which is known to bind (A/T)GATA(A/G) motifs in DNA and the N-finger, which is important for interacting with co-regulatory proteins such as Friend of GATA (FOG). We now show that, like the C-finger, the N-finger of GATA-1 is also capable of binding DNA but recognizes distinct sequences with the core GATC. We demonstrate that the GATA-1 N-finger can bind these sequences in vitro and that in cellular assays, GATA-1 can activate promoters containing GATC motifs. Experiments with mutant GATA-1 proteins confirm the importance of the N-finger, as the C-finger is not required for transactivation from GATC sites. Recently four naturally occurring mutations in GATA-1 have been shown to be associated with familial blood disorders. These mutations all map to the N-finger domain. We have investigated the effect of these mutations on the recognition of GATC sites by the N-finger and show that one mutation R216Q abolishes DNA binding, whereas the others have only minor effects.},\n bibtype = {article},\n author = {Newton, A. and Mackay, J. and Crossley, M.},\n doi = {10.1074/jbc.M106256200},\n journal = {Journal of Biological Chemistry},\n number = {38}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The mammalian transcription factor GATA-1 is required for normal erythroid and megakaryocytic development. GATA-1 contains two zinc fingers, the C-terminal finger, which is known to bind (A/T)GATA(A/G) motifs in DNA and the N-finger, which is important for interacting with co-regulatory proteins such as Friend of GATA (FOG). We now show that, like the C-finger, the N-finger of GATA-1 is also capable of binding DNA but recognizes distinct sequences with the core GATC. We demonstrate that the GATA-1 N-finger can bind these sequences in vitro and that in cellular assays, GATA-1 can activate promoters containing GATC motifs. Experiments with mutant GATA-1 proteins confirm the importance of the N-finger, as the C-finger is not required for transactivation from GATC sites. Recently four naturally occurring mutations in GATA-1 have been shown to be associated with familial blood disorders. These mutations all map to the N-finger domain. We have investigated the effect of these mutations on the recognition of GATC sites by the N-finger and show that one mutation R216Q abolishes DNA binding, whereas the others have only minor effects.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_2000\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_2000')\"\n      onkeypress=\"toggleGroup('group_2000')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>2000</span>\n      <span class=\"bibbase_group_count\">\n        \n        (6)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"liew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'liew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000', 'https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X', event);\">\n    Solution Structures of Two CCHC Zinc Fingers from the FOG Family Protein U-Shaped that Mediate Protein–Protein Interactions.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Liew,  C., K.; Kowalski,  K.; Fox,  A., H.; Newton,  A.; Sharpe,  B., K.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 8(11): 1157-1166. 11 2000.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X\"\n     onclick=\"log_download('liew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000', 'https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Solution Structures of Two CCHC Zinc Fingers from the FOG Family Protein U-Shaped that Mediate Protein–Protein Interactions [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0969-2126(00)00527-X\"\n     onclick=\"log_download('liew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000', 'http://doi.org/10.1016/S0969-2126(00)00527-X', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/liew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('liew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution Structures of Two CCHC Zinc Fingers from the FOG Family Protein U-Shaped that Mediate Protein–Protein Interactions},\n type = {article},\n year = {2000},\n keywords = {GATA-1,Structure,Transcription,U-shaped,Zinc fingers},\n pages = {1157-1166},\n volume = {8},\n websites = {https://linkinghub.elsevier.com/retrieve/pii/S096921260000527X},\n month = {11},\n id = {ee4bb2bf-b119-390f-a5c0-747836832737},\n created = {2020-12-17T05:29:58.497Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:58.497Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Liew2000},\n source_type = {ARTICLE},\n notes = {cited By 34},\n private_publication = {false},\n abstract = {Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.},\n bibtype = {article},\n author = {Liew, Chu Kong and Kowalski, Kasper and Fox, Archa H. and Newton, Anthea and Sharpe, Belinda K. and Crossley, Merlin and Mackay, Joel P.},\n doi = {10.1016/S0969-2126(00)00527-X},\n journal = {Structure},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"newton-sharped-kwan-mackay-crossley-thetransactivationdomainwithincysteinehistidinerichregion1ofcbpcomprisestwonovelzincbindingmodules-2000\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The transactivation domain within cysteine/histidine-rich region 1 of CBP comprises two novel zinc-binding modules.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Newton,  A.; Sharped,  B.; Kwan,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 275(20): 15128-15134.  2000.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M910396199\"\n     onclick=\"log_download('newton-sharped-kwan-mackay-crossley-thetransactivationdomainwithincysteinehistidinerichregion1ofcbpcomprisestwonovelzincbindingmodules-2000', 'http://doi.org/10.1074/jbc.M910396199', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/newton-sharped-kwan-mackay-crossley-thetransactivationdomainwithincysteinehistidinerichregion1ofcbpcomprisestwonovelzincbindingmodules-2000\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('newton-sharped-kwan-mackay-crossley-thetransactivationdomainwithincysteinehistidinerichregion1ofcbpcomprisestwonovelzincbindingmodules-2000')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The transactivation domain within cysteine/histidine-rich region 1 of CBP comprises two novel zinc-binding modules},\n type = {article},\n year = {2000},\n pages = {15128-15134},\n volume = {275},\n id = {912798e3-760b-3e6e-bdd0-d9324be5e544},\n created = {2023-01-10T01:46:37.833Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:37.833Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {cAMP-response element-binding protein-binding protein (CBP) is a transcriptional coactivator that interacts with a number of DNA-binding proteins and cofactor proteins involved in the regulation of transcription. Relatively little is known about the structure of CBP, but it has been noted that it contains three domains that are rich in cysteine and histidine (CH1, CH2, and CH3). The sequence of CH2 conforms to that of a leukemia-associated protein domain (PHD finger), and it has been postulated that this and both CH1 and CH3 may be zinc finger domains. This has not, however, been demonstrated experimentally. We have studied CH1 and show that it is composed of two novel zinc-binding modules, which we term &#x27;zinc bundles.&#x27; Each bundle contains the sequence Cys-X4-Cys-X8-His-X3-Cys, and we show that a synthetic peptide comprising one zinc bundle from CH1 can fold in a zinc- dependent manner. CH3 also appears to contain two zinc bundles, one with the variant sequence Cys-X2-Cys-X9-His-X3-Cys, and we demonstrate that this variant motif also undergoes Zn(II)-induced folding. CH1 acts as a transcriptional activation domain in cellular assays. We show that mutations in any of the four zinc-chelating residues in either zinc bundle of CH1 significantly impair this activity and that these mutations also interfere with certain protein-protein interactions mediated by CH1. Our results indicate that CBP is a genuine zinc-binding protein and introduce zinc bundles as novel protein interaction domains.},\n bibtype = {article},\n author = {Newton, A.L. and Sharped, B.K. and Kwan, A. and Mackay, J.P. and Crossley, M.},\n doi = {10.1074/jbc.M910396199},\n journal = {Journal of Biological Chemistry},\n number = {20}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  cAMP-response element-binding protein-binding protein (CBP) is a transcriptional coactivator that interacts with a number of DNA-binding proteins and cofactor proteins involved in the regulation of transcription. Relatively little is known about the structure of CBP, but it has been noted that it contains three domains that are rich in cysteine and histidine (CH1, CH2, and CH3). The sequence of CH2 conforms to that of a leukemia-associated protein domain (PHD finger), and it has been postulated that this and both CH1 and CH3 may be zinc finger domains. This has not, however, been demonstrated experimentally. We have studied CH1 and show that it is composed of two novel zinc-binding modules, which we term 'zinc bundles.' Each bundle contains the sequence Cys-X4-Cys-X8-His-X3-Cys, and we show that a synthetic peptide comprising one zinc bundle from CH1 can fold in a zinc- dependent manner. CH3 also appears to contain two zinc bundles, one with the variant sequence Cys-X2-Cys-X9-His-X3-Cys, and we demonstrate that this variant motif also undergoes Zn(II)-induced folding. CH1 acts as a transcriptional activation domain in cellular assays. We show that mutations in any of the four zinc-chelating residues in either zinc bundle of CH1 significantly impair this activity and that these mutations also interfere with certain protein-protein interactions mediated by CH1. Our results indicate that CBP is a genuine zinc-binding protein and introduce zinc bundles as novel protein interaction domains.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"matthews-young-tucker-mackay-thecoreoftherespiratorysyncytialvirusfusionproteinisatrimericcoiledcoil-2000\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The core of the respiratory syncytial virus fusion protein is a trimeric coiled coil.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Matthews,  J.; Young,  T.; Tucker,  S.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Virology</i>, 74(13): 5911-5920.  2000.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1128/JVI.74.13.5911-5920.2000\"\n     onclick=\"log_download('matthews-young-tucker-mackay-thecoreoftherespiratorysyncytialvirusfusionproteinisatrimericcoiledcoil-2000', 'http://doi.org/10.1128/JVI.74.13.5911-5920.2000', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/matthews-young-tucker-mackay-thecoreoftherespiratorysyncytialvirusfusionproteinisatrimericcoiledcoil-2000\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('matthews-young-tucker-mackay-thecoreoftherespiratorysyncytialvirusfusionproteinisatrimericcoiledcoil-2000')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The core of the respiratory syncytial virus fusion protein is a trimeric coiled coil},\n type = {article},\n year = {2000},\n pages = {5911-5920},\n volume = {74},\n id = {82505942-a661-3722-bf77-e61b0007b83b},\n created = {2023-01-10T01:46:38.745Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:38.745Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Entry into the host cell by enveloped viruses is mediated by fusion (F) or transmembrane glycoproteins. Many of these proteins share a fold comprising a trimer of antiparallel coiled-coil heterodimers, where the heterodimers are formed by two discontinuous heptad repeat motifs within the proteolytically processed chain. The F protein of human respiratory syncytial virus (RSV; the major cause of lower respiratory tract infections in infants) contains two corresponding regions that are predicted to form coiled coils (HR1 and HR2), together with a third predicted heptad repeat (HR3) located in a nonhomologous position. In order to probe the structures of these three domains and ascertain the nature of the interactions between them, we have studied the isolated HR1, HR2, and HR3 domains of RSV F by using a range of biophysical techniques, including circular dichroism, nuclear magnetic resonance spectroscopy, and sedimentation equilibrium. HR1 forms a symmetrical, trimeric coiled coil in solution (K3 ≃ 2.2 x 1011 M-2) which interacts with HR2 to form a 3:3 hexamer. The HR1-HR2 interaction domains have been mapped using limited proteolysis, reversed-phase high- performance liquid chromatography, and electrospray-mass spectrometry. HR2 in isolation exists as a largely unstructured monomer, although it exhibits a tendency to form aggregates with β-sheet-like characteristics. Only a small increase in α-helical content was observed upon the formation of the hexamer. This suggests that the RSV F glycoprotein contains a domain that closely resembles the core structure of the simian parainfluenza virus 5 fusion protein (K. A. Baker, R. E. Dutch, R. A. Lamb, and T. S. Jardetzky, Mol. Cell 3:309-319, 1999). Finally, HR3 forms weak α-helical homodimers that do not appear to interact with HR1, HR2, or the HR1-HR2 complex. The results of these studies support the idea that viral fusion proteins have a common core architecture.},\n bibtype = {article},\n author = {Matthews, J.M. and Young, T.F. and Tucker, S.P. and Mackay, J.P.},\n doi = {10.1128/JVI.74.13.5911-5920.2000},\n journal = {Journal of Virology},\n number = {13}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Entry into the host cell by enveloped viruses is mediated by fusion (F) or transmembrane glycoproteins. Many of these proteins share a fold comprising a trimer of antiparallel coiled-coil heterodimers, where the heterodimers are formed by two discontinuous heptad repeat motifs within the proteolytically processed chain. The F protein of human respiratory syncytial virus (RSV; the major cause of lower respiratory tract infections in infants) contains two corresponding regions that are predicted to form coiled coils (HR1 and HR2), together with a third predicted heptad repeat (HR3) located in a nonhomologous position. In order to probe the structures of these three domains and ascertain the nature of the interactions between them, we have studied the isolated HR1, HR2, and HR3 domains of RSV F by using a range of biophysical techniques, including circular dichroism, nuclear magnetic resonance spectroscopy, and sedimentation equilibrium. HR1 forms a symmetrical, trimeric coiled coil in solution (K3 ≃ 2.2 x 1011 M-2) which interacts with HR2 to form a 3:3 hexamer. The HR1-HR2 interaction domains have been mapped using limited proteolysis, reversed-phase high- performance liquid chromatography, and electrospray-mass spectrometry. HR2 in isolation exists as a largely unstructured monomer, although it exhibits a tendency to form aggregates with β-sheet-like characteristics. Only a small increase in α-helical content was observed upon the formation of the hexamer. This suggests that the RSV F glycoprotein contains a domain that closely resembles the core structure of the simian parainfluenza virus 5 fusion protein (K. A. Baker, R. E. Dutch, R. A. Lamb, and T. S. Jardetzky, Mol. Cell 3:309-319, 1999). Finally, HR3 forms weak α-helical homodimers that do not appear to interact with HR1, HR2, or the HR1-HR2 complex. The results of these studies support the idea that viral fusion proteins have a common core architecture.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"chukongliew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Solution structures of two CCHC zinc fingers from the FOG family protein U-shaped that mediate protein-protein interactions.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Chu Kong Liew; Kowalski,  K.; Fox,  A.; Newton,  A.; Sharpe,  B.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 8(11): 1157-1166.  2000.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0969-2126(00)00527-X\"\n     onclick=\"log_download('chukongliew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000', 'http://doi.org/10.1016/S0969-2126(00)00527-X', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/chukongliew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('chukongliew-kowalski-fox-newton-sharpe-crossley-mackay-solutionstructuresoftwocchczincfingersfromthefogfamilyproteinushapedthatmediateproteinproteininteractions-2000')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Solution structures of two CCHC zinc fingers from the FOG family protein U-shaped that mediate protein-protein interactions},\n type = {article},\n year = {2000},\n pages = {1157-1166},\n volume = {8},\n id = {4220c92e-ca96-3078-a476-5f478fe44cdb},\n created = {2023-01-10T01:46:39.669Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:39.669Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.},\n bibtype = {article},\n author = {Chu Kong Liew, undefined and Kowalski, K. and Fox, A.H. and Newton, A. and Sharpe, B.K. and Crossley, M. and Mackay, J.P.},\n doi = {10.1016/S0969-2126(00)00527-X},\n journal = {Structure},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: Zinc finger domains have traditionally been regarded as sequence-specific DNA binding motifs. However, recent evidence indicates that many zinc fingers mediate specific protein-protein interactions. For instance, several zinc fingers from FOG family proteins have been shown to interact with the N-terminal zinc finger of GATA-1. Results: We have used NMR spectroscopy to determine the first structures of two FOG family zinc fingers that are involved in protein-protein interactions: fingers 1 and 9 from U-shaped. These fingers resemble classical TFIIIA-like zinc fingers, with the exception of an unusual extended portion of the polypeptide backbone prior to the fourth zinc ligand. [15N,1H]-HSQC titrations have been used to define the GATA binding surface of USH-F1, and comparison with other FOG family proteins indicates that the recognition mechanism is conserved across species. The surface of FOG-type fingers that interacts with GATA-1 overlaps substantially with the surface through which classical fingers typically recognize DNA. This suggests that these fingers could not contact both GATA and DNA simultaneously. In addition, results from NMR, gel filtration, and sedimentation equilibrium experiments suggest that the interactions are of moderate affinity. Conclusions: Our results demonstrate unequivocally that zinc fingers comprising the classical ββα fold are capable of mediating specific contacts between proteins. The existence of this alternative function has implications for the prediction of protein function from sequence data and for the evolution of protein function.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"tchan-choy-mackay-lyons-bains-weiss-interfacialasparagineresidueswithinanamidetetradcontributetomaxhelixloophelixleucinezipperhomodimerstability-2000\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Interfacial asparagine residues within an amide tetrad contribute to Max helix-loop-helix leucine zipper homodimer stability.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Tchan,  M.; Choy,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Lyons,  A.; Bains,  N.;  and Weiss,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 275(48): 37454-37461.  2000.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.M004264200\"\n     onclick=\"log_download('tchan-choy-mackay-lyons-bains-weiss-interfacialasparagineresidueswithinanamidetetradcontributetomaxhelixloophelixleucinezipperhomodimerstability-2000', 'http://doi.org/10.1074/jbc.M004264200', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/tchan-choy-mackay-lyons-bains-weiss-interfacialasparagineresidueswithinanamidetetradcontributetomaxhelixloophelixleucinezipperhomodimerstability-2000\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('tchan-choy-mackay-lyons-bains-weiss-interfacialasparagineresidueswithinanamidetetradcontributetomaxhelixloophelixleucinezipperhomodimerstability-2000')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Interfacial asparagine residues within an amide tetrad contribute to Max helix-loop-helix leucine zipper homodimer stability},\n type = {article},\n year = {2000},\n pages = {37454-37461},\n volume = {275},\n id = {44466ab6-2aa6-3104-b086-fd106c65ff76},\n created = {2023-01-10T01:46:40.592Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:40.592Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The transcription factor Max is the obligate dimerization partner of the Myc oncoprotein. The pivotal role of Max within the Myc regulatory network is dependent upon its ability to dimerize via the helix-loop-helix leucine zipper domain. The Max homodimer contains a tetrad of polar residues at the interface of the leucine zipper domain. A conserved interfacial Asn residue at an equivalent position in two other leucine zipper proteins has been shown to decrease homodimer stability. The unusual arrangement of this Gln-Asn/Gln&#x27;-Asn&#x27; tetrad prompted us to investigate whether Asn92 plays a similar role in destabilizing the Max homodimer. This residue was sequentially replaced with aliphatic and charged residues. Thermal denaturation, redox time course and analytical ultracentrifugation studies show that the N92V mutation does not increase homodimer stability. Replacing this residue with negatively charged side chains in N92D and N92E destabilizes the mutant homodimer. Further replacement of Gln91 indicated that H bonding between Gln91 and Asn92 residues is not significant to the stability of the native protein. These data collectively demonstrate the central role of Asn92 in homodimer interactions. Molecular modelling studies illustrate the favorable packing of the native Asn residue at the dimer interface compared with that of the mutant Max peptides.},\n bibtype = {article},\n author = {Tchan, M.C. and Choy, K.J. and Mackay, J.P. and Lyons, A.T.L. and Bains, N.P.S. and Weiss, A.S.},\n doi = {10.1074/jbc.M004264200},\n journal = {Journal of Biological Chemistry},\n number = {48}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The transcription factor Max is the obligate dimerization partner of the Myc oncoprotein. The pivotal role of Max within the Myc regulatory network is dependent upon its ability to dimerize via the helix-loop-helix leucine zipper domain. The Max homodimer contains a tetrad of polar residues at the interface of the leucine zipper domain. A conserved interfacial Asn residue at an equivalent position in two other leucine zipper proteins has been shown to decrease homodimer stability. The unusual arrangement of this Gln-Asn/Gln'-Asn' tetrad prompted us to investigate whether Asn92 plays a similar role in destabilizing the Max homodimer. This residue was sequentially replaced with aliphatic and charged residues. Thermal denaturation, redox time course and analytical ultracentrifugation studies show that the N92V mutation does not increase homodimer stability. Replacing this residue with negatively charged side chains in N92D and N92E destabilizes the mutant homodimer. Further replacement of Gln91 indicated that H bonding between Gln91 and Asn92 residues is not significant to the stability of the native protein. These data collectively demonstrate the central role of Asn92 in homodimer interactions. Molecular modelling studies illustrate the favorable packing of the native Asn residue at the dimer interface compared with that of the mutant Max peptides.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"matthews-kowalski-liew-sharpe-fox-crossley-mackay-aclassofzincfingersinvolvedinproteinproteininteractions-2000\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  A class of zinc fingers involved in protein-protein interactions.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Matthews,  J.; Kowalski,  K.; Liew,  C.; Sharpe,  B.; Fox,  A.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>European Journal of Biochemistry</i>, 267(4): 1030-1038.  2000.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1046/j.1432-1327.2000.01095.x\"\n     onclick=\"log_download('matthews-kowalski-liew-sharpe-fox-crossley-mackay-aclassofzincfingersinvolvedinproteinproteininteractions-2000', 'http://doi.org/10.1046/j.1432-1327.2000.01095.x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/matthews-kowalski-liew-sharpe-fox-crossley-mackay-aclassofzincfingersinvolvedinproteinproteininteractions-2000\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('matthews-kowalski-liew-sharpe-fox-crossley-mackay-aclassofzincfingersinvolvedinproteinproteininteractions-2000')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {A class of zinc fingers involved in protein-protein interactions},\n type = {article},\n year = {2000},\n pages = {1030-1038},\n volume = {267},\n id = {3aade15c-920e-3fd6-9d2d-2812368f8f88},\n created = {2023-01-10T01:46:41.497Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:41.497Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Zinc fingers (ZnFs) are extremely common protein domains. Several classes of ZnFs are distinguished by the nature and spacing of their zinc- coordinating residues. While the structure and function of some ZnFs are well characterized, many others have been identified only through their amino acid sequence. A number of proteins contain a conserved C-X2-C-X12-H-X1-5-C sequence, which is similar to the spacing observed for the &#x27;classic&#x27; CCHH ZnFs. Although these domains have been implicated in protein-protein (and not protein-nucleic acid) interactions, nothing is known about their structure or function at a molecular level. Here, we address this problem through the expression and biophysical characterization of several CCHC-type zinc fingers from the erythroid transcription factor FOG and the related Drosophila protein U-shaped. Each of these domains does indeed fold in a zinc-dependent fashion, coordinating the metal in a tetrahedral manner through the sidechains of one histidine and three cysteine residues, and forming extremely thermostable structures. Analysis of CD spectra suggests an overall fold similar to that of the CCHH fingers, and indeed a point mutant of FOG-F1 in which the final cysteine residue is replaced by histidine remains capable of folding. However, the CCHC (as opposed to CCHH) motif is a prerequisite for GATA-1 binding activity, demonstrating that CCHC and CCHH topologies are not interchangeable. This demonstration that members of a structurally distinct subclass of genuine zinc finger domains are involved in the mediation of protein-protein interactions has implications for the prediction of protein function from nucleotide sequences.},\n bibtype = {article},\n author = {Matthews, J.M. and Kowalski, K. and Liew, C.K. and Sharpe, B.K. and Fox, A.H. and Crossley, M. and Mackay, J.P.},\n doi = {10.1046/j.1432-1327.2000.01095.x},\n journal = {European Journal of Biochemistry},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Zinc fingers (ZnFs) are extremely common protein domains. Several classes of ZnFs are distinguished by the nature and spacing of their zinc- coordinating residues. While the structure and function of some ZnFs are well characterized, many others have been identified only through their amino acid sequence. A number of proteins contain a conserved C-X2-C-X12-H-X1-5-C sequence, which is similar to the spacing observed for the 'classic' CCHH ZnFs. Although these domains have been implicated in protein-protein (and not protein-nucleic acid) interactions, nothing is known about their structure or function at a molecular level. Here, we address this problem through the expression and biophysical characterization of several CCHC-type zinc fingers from the erythroid transcription factor FOG and the related Drosophila protein U-shaped. Each of these domains does indeed fold in a zinc-dependent fashion, coordinating the metal in a tetrahedral manner through the sidechains of one histidine and three cysteine residues, and forming extremely thermostable structures. Analysis of CD spectra suggests an overall fold similar to that of the CCHH fingers, and indeed a point mutant of FOG-F1 in which the final cysteine residue is replaced by histidine remains capable of folding. However, the CCHC (as opposed to CCHH) motif is a prerequisite for GATA-1 binding activity, demonstrating that CCHC and CCHH topologies are not interchangeable. This demonstration that members of a structurally distinct subclass of genuine zinc finger domains are involved in the mediation of protein-protein interactions has implications for the prediction of protein function from nucleotide sequences.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_1999\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_1999')\"\n      onkeypress=\"toggleGroup('group_1999')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>1999</span>\n      <span class=\"bibbase_group_count\">\n        \n        (6)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/10212985\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999', 'http://www.ncbi.nlm.nih.gov/pubmed/10212985', event);\">\n    The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kowalski,  K.; Czolij,  R.; King,  G., F.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biomolecular NMR</i>, 13(3): 249-62. 3 1999.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.ncbi.nlm.nih.gov/pubmed/10212985\"\n     onclick=\"log_download('kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999', 'http://www.ncbi.nlm.nih.gov/pubmed/10212985', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG. [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/https://doi.org/10.1023/a:1008309602929\"\n     onclick=\"log_download('kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999', 'http://doi.org/https://doi.org/10.1023/a:1008309602929', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG.},\n type = {article},\n year = {1999},\n keywords = {GATA-1,Protein-protein interactions,Solution structure,Zinc finger},\n pages = {249-62},\n volume = {13},\n websites = {http://www.ncbi.nlm.nih.gov/pubmed/10212985},\n month = {3},\n id = {1bc4df8b-a093-3a22-ba88-996d8029dfd5},\n created = {2020-12-17T05:29:56.853Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:56.853Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Kowalski1999},\n source_type = {ARTICLE},\n notes = {cited By 48},\n private_publication = {false},\n abstract = {Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted beta-hairpins and a single alpha-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA-1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.},\n bibtype = {article},\n author = {Kowalski, K. and Czolij, R. and King, G. F. and Crossley, M. and Mackay, J. P.},\n doi = {https://doi.org/10.1023/a:1008309602929},\n journal = {Journal of Biomolecular NMR},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted beta-hairpins and a single alpha-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA-1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Kowalski,  K.; Czolij,  R.; King,  G.; Crossley,  M.;  and <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biomolecular NMR</i>, 13(3): 249-262.  1999.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1023/A:1008309602929\"\n     onclick=\"log_download('kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999', 'http://doi.org/10.1023/A:1008309602929', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('kowalski-czolij-king-crossley-mackay-thesolutionstructureofthenterminalzincfingerofgata1revealsaspecificbindingfaceforthetranscriptionalcofactorfog-1999')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG},\n type = {article},\n year = {1999},\n pages = {249-262},\n volume = {13},\n id = {649a742c-0f2a-3975-8e3d-1abdb42ec917},\n created = {2023-01-10T01:46:42.416Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:42.416Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted β-hairpins and a single α-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA- 1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.},\n bibtype = {article},\n author = {Kowalski, K. and Czolij, R. and King, G.F. and Crossley, M. and Mackay, J.P.},\n doi = {10.1023/A:1008309602929},\n journal = {Journal of Biomolecular NMR},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional 1H/15N NMR spectroscopy, and we use 1H/15N spin relaxation measurements to investigate its backbone dynamics. The structure consists of two distorted β-hairpins and a single α-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NF. Notably, this face is well removed from the putative DNA-binding face of the NF, an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA- 1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"king-rowland-pan-mackay-mullen-rothfield-thedimerizationandtopologicalspecificityfunctionsofmineresideinastructurallyautonomouscterminaldomain-1999\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The dimerization and topological specificity functions of MinE reside in a structurally autonomous C-terminal domain.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  King,  G.; Rowland,  S.; Pan,  B.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Mullen,  G.;  and Rothfield,  L.\n</span>\n\n  \n\n</span>\n\n  <i>Molecular Microbiology</i>, 31(4): 1161-1169.  1999.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1046/j.1365-2958.1999.01256.x\"\n     onclick=\"log_download('king-rowland-pan-mackay-mullen-rothfield-thedimerizationandtopologicalspecificityfunctionsofmineresideinastructurallyautonomouscterminaldomain-1999', 'http://doi.org/10.1046/j.1365-2958.1999.01256.x', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/king-rowland-pan-mackay-mullen-rothfield-thedimerizationandtopologicalspecificityfunctionsofmineresideinastructurallyautonomouscterminaldomain-1999\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('king-rowland-pan-mackay-mullen-rothfield-thedimerizationandtopologicalspecificityfunctionsofmineresideinastructurallyautonomouscterminaldomain-1999')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The dimerization and topological specificity functions of MinE reside in a structurally autonomous C-terminal domain},\n type = {article},\n year = {1999},\n pages = {1161-1169},\n volume = {31},\n id = {cd93d5ad-9177-3466-a3fb-e5c4f562db3d},\n created = {2023-01-10T01:46:43.487Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:43.487Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Correct placement of the division septum in Escherichia coli requires the co-ordinated action of three proteins, MinC, MinD and MinE. MinC and MinD interact to form a non-specific division inhibitor that blocks septation at all potential division sites. MinE is able to antagonize MinCD in a topologically sensitive manner, as it restricts MinCD activity to the unwanted division sites at the cell poles. Here, we show that the topological specificity function of MinE residues in a structurally autonomous, trypsin-resistant domain comprising residues 31-88. Nuclear magnetic resonance (NMR) and circular dichroic spectroscopy indicate that this domain includes both α and β secondary structure, while analytical ultracentrifugation reveals that it also contains a region responsible for MinE homodimerization. While trypsin digestion indicates that the anti-MinCD domain of MinE (residues 1-22) does not form a tightly folded structural domain, NMR analysis of a peptide corresponding to MinE1-22 indicates that this region forms a nascent helix in which the peptide rapidly interconverts between disordered (random coil) and α-helical conformations. This suggests that the N-terminal region of MinE may be poised to adopt an α-helical conformation when it interacts with the target of its anti-MinCD activity, presumably MinD.},\n bibtype = {article},\n author = {King, G.F. and Rowland, S.L. and Pan, B. and Mackay, J.P. and Mullen, G.P. and Rothfield, L.I.},\n doi = {10.1046/j.1365-2958.1999.01256.x},\n journal = {Molecular Microbiology},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Correct placement of the division septum in Escherichia coli requires the co-ordinated action of three proteins, MinC, MinD and MinE. MinC and MinD interact to form a non-specific division inhibitor that blocks septation at all potential division sites. MinE is able to antagonize MinCD in a topologically sensitive manner, as it restricts MinCD activity to the unwanted division sites at the cell poles. Here, we show that the topological specificity function of MinE residues in a structurally autonomous, trypsin-resistant domain comprising residues 31-88. Nuclear magnetic resonance (NMR) and circular dichroic spectroscopy indicate that this domain includes both α and β secondary structure, while analytical ultracentrifugation reveals that it also contains a region responsible for MinE homodimerization. While trypsin digestion indicates that the anti-MinCD domain of MinE (residues 1-22) does not form a tightly folded structural domain, NMR analysis of a peptide corresponding to MinE1-22 indicates that this region forms a nascent helix in which the peptide rapidly interconverts between disordered (random coil) and α-helical conformations. This suggests that the N-terminal region of MinE may be poised to adopt an α-helical conformation when it interacts with the target of its anti-MinCD activity, presumably MinD.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"zerella-evans-ionides-packman-trotter-mackay-williams-autonomousfoldingofapeptidecorrespondingtothenterminalhairpinfromubiquitin-1999\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Autonomous folding of a peptide corresponding to the N-terminal β- hairpin from ubiquitin.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Zerella,  R.; Evans,  P.; Ionides,  J.; Packman,  L.; Trotter,  B.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Williams,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Protein Science</i>, 8(6): 1320-1331.  1999.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1110/ps.8.6.1320\"\n     onclick=\"log_download('zerella-evans-ionides-packman-trotter-mackay-williams-autonomousfoldingofapeptidecorrespondingtothenterminalhairpinfromubiquitin-1999', 'http://doi.org/10.1110/ps.8.6.1320', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/zerella-evans-ionides-packman-trotter-mackay-williams-autonomousfoldingofapeptidecorrespondingtothenterminalhairpinfromubiquitin-1999\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('zerella-evans-ionides-packman-trotter-mackay-williams-autonomousfoldingofapeptidecorrespondingtothenterminalhairpinfromubiquitin-1999')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Autonomous folding of a peptide corresponding to the N-terminal β- hairpin from ubiquitin},\n type = {article},\n year = {1999},\n pages = {1320-1331},\n volume = {8},\n id = {2fa2cd79-9bc1-3f5c-8753-6943e5016588},\n created = {2023-01-10T01:46:44.399Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:44.399Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The N-terminal 17 residues of ubiquitin have been shown by 1H NMR to fold autonomously into a β-hairpin structure in aqueous solution. This structure has a specific, native-like register, though side-chain contacts differ in detail from those observed in the intact protein. An autonomously folding hairpin has previously been identified in the case of streptococcal protein G, which is structurally homologous with ubiquitin, but remarkably, the two are not in topologically equivalent positions in the fold. This suggests that the organization of folding may be quite different for proteins sharing similar tertiary structures. Two smaller peptides have also been studied, corresponding to the isolated arms of the N-terminal hairpin of ubiquitin, and significant differences from simple random coil predictions observed in the spectra of these subfragments, suggestive of significant limitation of the backbone conformational space sampled, presumably as a consequence of the strongly β-structure favoring composition of the sequences. This illustrates the ability of local sequence elements to express a propensity for β-structure even in the absence of actual sheet formation. Attempts were made to estimate the population of the folded state of the hairpin, in terms of a simple two-state folding model. Using published &#x27;random coil&#x27; values to model the unfolded state, and values derived from native ubiquitin for the putative unique, folded state, it was found that the apparent population varied widely for different residues and with different NMR parameters. Use of the spectra of the subfragment peptides to provide a more realistic model of the unfolded state led to better agreement in the estimates that could be obtained from chemical shift and coupling constant measurements, while making it clear that some other approaches to population estimation could not give meaningful results, because of the tendency to populate the β-region of conformational space even in the absence of the hairpin structure.},\n bibtype = {article},\n author = {Zerella, R. and Evans, P.A. and Ionides, J.M.C. and Packman, L.C. and Trotter, B.W. and Mackay, J.P. and Williams, D.H.},\n doi = {10.1110/ps.8.6.1320},\n journal = {Protein Science},\n number = {6}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The N-terminal 17 residues of ubiquitin have been shown by 1H NMR to fold autonomously into a β-hairpin structure in aqueous solution. This structure has a specific, native-like register, though side-chain contacts differ in detail from those observed in the intact protein. An autonomously folding hairpin has previously been identified in the case of streptococcal protein G, which is structurally homologous with ubiquitin, but remarkably, the two are not in topologically equivalent positions in the fold. This suggests that the organization of folding may be quite different for proteins sharing similar tertiary structures. Two smaller peptides have also been studied, corresponding to the isolated arms of the N-terminal hairpin of ubiquitin, and significant differences from simple random coil predictions observed in the spectra of these subfragments, suggestive of significant limitation of the backbone conformational space sampled, presumably as a consequence of the strongly β-structure favoring composition of the sequences. This illustrates the ability of local sequence elements to express a propensity for β-structure even in the absence of actual sheet formation. Attempts were made to estimate the population of the folded state of the hairpin, in terms of a simple two-state folding model. Using published 'random coil' values to model the unfolded state, and values derived from native ubiquitin for the putative unique, folded state, it was found that the apparent population varied widely for different residues and with different NMR parameters. Use of the spectra of the subfragment peptides to provide a more realistic model of the unfolded state led to better agreement in the estimates that could be obtained from chemical shift and coupling constant measurements, while making it clear that some other approaches to population estimation could not give meaningful results, because of the tendency to populate the β-region of conformational space even in the absence of the hairpin structure.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"fox-liew-holmes-kowalski-mackay-crossley-transcriptionalcofactorsofthefogfamilyinteractwithgataproteinsbymeansofmultiplezincfingers-1999\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Transcriptional cofactors of the FOG family interact with GATA proteins by means of multiple zinc fingers.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Fox,  A.; Liew,  C.; Holmes,  M.; Kowalski,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>EMBO Journal</i>, 18(10): 2812-2822.  1999.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1093/emboj/18.10.2812\"\n     onclick=\"log_download('fox-liew-holmes-kowalski-mackay-crossley-transcriptionalcofactorsofthefogfamilyinteractwithgataproteinsbymeansofmultiplezincfingers-1999', 'http://doi.org/10.1093/emboj/18.10.2812', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/fox-liew-holmes-kowalski-mackay-crossley-transcriptionalcofactorsofthefogfamilyinteractwithgataproteinsbymeansofmultiplezincfingers-1999\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('fox-liew-holmes-kowalski-mackay-crossley-transcriptionalcofactorsofthefogfamilyinteractwithgataproteinsbymeansofmultiplezincfingers-1999')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Transcriptional cofactors of the FOG family interact with GATA proteins by means of multiple zinc fingers},\n type = {article},\n year = {1999},\n pages = {2812-2822},\n volume = {18},\n id = {008d2a64-1e9c-3d79-90e8-1d4d6afcbc71},\n created = {2023-01-10T01:46:45.293Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:45.293Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Friend of GATA-1 (FOG-1) is a zinc finger protein that has been shown to interact physically with the erythroid DNA-binding protein GATA-1 and modulate its transcriptional activity. Recently, two new members of the FOG family have been identified: a mammalian protein, FOG-2, that also associates with GATA-1 and other mammalian GATA factors; and U-shaped, a Drosophila protein that interacts with the Drosophila GATA protein Pannier. FOG proteins contain multiple zinc fingers and it has been shown previously that the sixth finger of FOG-1 interacts specifically with the N-finger but not the C-finger of GATA-1. Here we show that fingers 1, 5 and 9 of FOG-1 also interact with the N-finger of GATA-1 and that FOG-2 and U-shaped also contain multiple GATA-interacting fingers. We define the key contact residues and show that these residues are highly conserved in GATA-interacting fingers. We examine the effect of selectively mutating the four interacting fingers of FOG-1 and show that each contributes to FOG-1&#x27;s ability to modulate GATA-1 activity. Finally, we show that FOG-1 can repress GATA-1-mediated activation and present evidence that this ability involves the recently described CtBP co-repressor proteins that recognize all known FOG proteins.},\n bibtype = {article},\n author = {Fox, A.H. and Liew, C. and Holmes, M. and Kowalski, K. and Mackay, J. and Crossley, M.},\n doi = {10.1093/emboj/18.10.2812},\n journal = {EMBO Journal},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Friend of GATA-1 (FOG-1) is a zinc finger protein that has been shown to interact physically with the erythroid DNA-binding protein GATA-1 and modulate its transcriptional activity. Recently, two new members of the FOG family have been identified: a mammalian protein, FOG-2, that also associates with GATA-1 and other mammalian GATA factors; and U-shaped, a Drosophila protein that interacts with the Drosophila GATA protein Pannier. FOG proteins contain multiple zinc fingers and it has been shown previously that the sixth finger of FOG-1 interacts specifically with the N-finger but not the C-finger of GATA-1. Here we show that fingers 1, 5 and 9 of FOG-1 also interact with the N-finger of GATA-1 and that FOG-2 and U-shaped also contain multiple GATA-interacting fingers. We define the key contact residues and show that these residues are highly conserved in GATA-interacting fingers. We examine the effect of selectively mutating the four interacting fingers of FOG-1 and show that each contributes to FOG-1's ability to modulate GATA-1 activity. Finally, we show that FOG-1 can repress GATA-1-mediated activation and present evidence that this ability involves the recently described CtBP co-repressor proteins that recognize all known FOG proteins.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"crispino-lodish-mackay-orkin-useofalteredspecificitymutantstoprobeaspecificproteinproteininteractionindifferentiationthegata1fogcomplex-1999\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Use of altered specificity mutants to probe a specific protein-protein interaction in differentiation: The GATA-1:FOG complex.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Crispino,  J.; Lodish,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">MacKay,  J.</a>;  and Orkin,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Molecular Cell</i>, 3(2): 219-228.  1999.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S1097-2765(00)80312-3\"\n     onclick=\"log_download('crispino-lodish-mackay-orkin-useofalteredspecificitymutantstoprobeaspecificproteinproteininteractionindifferentiationthegata1fogcomplex-1999', 'http://doi.org/10.1016/S1097-2765(00)80312-3', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/crispino-lodish-mackay-orkin-useofalteredspecificitymutantstoprobeaspecificproteinproteininteractionindifferentiationthegata1fogcomplex-1999\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('crispino-lodish-mackay-orkin-useofalteredspecificitymutantstoprobeaspecificproteinproteininteractionindifferentiationthegata1fogcomplex-1999')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Use of altered specificity mutants to probe a specific protein-protein interaction in differentiation: The GATA-1:FOG complex},\n type = {article},\n year = {1999},\n pages = {219-228},\n volume = {3},\n id = {ed2ce0ef-2974-32de-ac4b-b652a7d61a3b},\n created = {2023-01-10T01:46:46.200Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:46.200Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {GATA-1 and FOG (Friend of GATA-1) are each essential for erythroid and megakaryocyte development. FOG, a zinc finger protein, interacts with the amino (N) finger of GATA-1 and cooperates with GATA-1 to promote differentiation. To determine whether this interaction is critical for GATA- 1 action, we selected GATA-1 mutants in yeast that fail to interact with FOG but retain normal DNA binding, as well a compensatory FOG mutant that restores interaction. These novel GATA-1 mutants do not promote erythroid differentiation of GATA-1- erythroid cells. Differentiation is rescued by the second-site FOG mutant. Thus, interaction of FOG with GATA-1 is essential for the function of GATA-1 in erythroid differentiation. These findings provide a paradigm for dissecting protein-protein associations involved in mammalian development.},\n bibtype = {article},\n author = {Crispino, J.D. and Lodish, M.B. and MacKay, J.P. and Orkin, S.H.},\n doi = {10.1016/S1097-2765(00)80312-3},\n journal = {Molecular Cell},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  GATA-1 and FOG (Friend of GATA-1) are each essential for erythroid and megakaryocyte development. FOG, a zinc finger protein, interacts with the amino (N) finger of GATA-1 and cooperates with GATA-1 to promote differentiation. To determine whether this interaction is critical for GATA- 1 action, we selected GATA-1 mutants in yeast that fail to interact with FOG but retain normal DNA binding, as well a compensatory FOG mutant that restores interaction. These novel GATA-1 mutants do not promote erythroid differentiation of GATA-1- erythroid cells. Differentiation is rescued by the second-site FOG mutant. Thus, interaction of FOG with GATA-1 is essential for the function of GATA-1 in erythroid differentiation. These findings provide a paradigm for dissecting protein-protein associations involved in mammalian development.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_1998\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_1998')\"\n      onkeypress=\"toggleGroup('group_1998')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>1998</span>\n      <span class=\"bibbase_group_count\">\n        \n        (5)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"diefenbach-mackay-armati-cunningham-thecterminalregionofthestalkdomainofubiquitoushumankinesinheavychaincontainsthebindingsiteforkinesinlightchain-1998\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The C-terminal region of the stalk domain of ubiquitous human kinesin heavy chain contains the binding site for kinesin light chain.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Diefenbach,  R.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Armati,  P.;  and Cunningham,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 37(47): 16663-16670.  1998.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi981163r\"\n     onclick=\"log_download('diefenbach-mackay-armati-cunningham-thecterminalregionofthestalkdomainofubiquitoushumankinesinheavychaincontainsthebindingsiteforkinesinlightchain-1998', 'http://doi.org/10.1021/bi981163r', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/diefenbach-mackay-armati-cunningham-thecterminalregionofthestalkdomainofubiquitoushumankinesinheavychaincontainsthebindingsiteforkinesinlightchain-1998\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('diefenbach-mackay-armati-cunningham-thecterminalregionofthestalkdomainofubiquitoushumankinesinheavychaincontainsthebindingsiteforkinesinlightchain-1998')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The C-terminal region of the stalk domain of ubiquitous human kinesin heavy chain contains the binding site for kinesin light chain},\n type = {article},\n year = {1998},\n pages = {16663-16670},\n volume = {37},\n id = {b73ee519-2d24-3501-8e20-4c8b4ead145e},\n created = {2023-01-10T01:46:47.102Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:47.102Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The motor protein kinesin is a heterotetramer composed of two heavy chains of ~120 kDa and two light chains of ~6 kDa protein. Kinesin motor activity is dependent on the presence of ATP and microtubules. The kinesin light chain-binding site in human kinesin heavy chain was determined by reconstituting in vitro a complex of recombinant heavy and light chains. The proteins expressed in bacteria included oligohistidine-tagged fragments of human ubiquitous kinesin heavy chain, spanning most of the stalk and all of the tail domain (amino acids 555-963); and untagged, essentially full-length human kinesin light chain (4-569) along with N-terminal (4-363)and C-terminal (364-569) light chain fragments. Heavy chain fragments were attached to Ni2+-charged beads and incubated with untagged light chain fragments. Analysis of eluted complexes by SDS-PAGE and immunoblotting mapped the light chain-binding site in heavy chain to amino acids 771-813, a region close to the C-terminal end of the heavy chain stalk domain. In addition, only the full-length and N-terminal kinesin light chain fragments bound to this heavy chain region. Within this heavy chain region are four highly conserved contiguous heptad repeats (775-802) which are predicted to form a tight α- helical coiled-coil interaction with the heptad repeat-containing N-terminus of the light chain, in particular region 106-152 of human light chain. This predicted hydrophobic, α-helical coiled-coil interaction is supported by both circular dichroism spectroscopy of the recombinant kinesin heavy chain fragment 771-963, which displays an α-helical content of 70%, and the resistance of the heavy/light chain interdiction to high salt (0.5 M).},\n bibtype = {article},\n author = {Diefenbach, R.J. and Mackay, J.P. and Armati, P.J. and Cunningham, A.L.},\n doi = {10.1021/bi981163r},\n journal = {Biochemistry},\n number = {47}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The motor protein kinesin is a heterotetramer composed of two heavy chains of ~120 kDa and two light chains of ~6 kDa protein. Kinesin motor activity is dependent on the presence of ATP and microtubules. The kinesin light chain-binding site in human kinesin heavy chain was determined by reconstituting in vitro a complex of recombinant heavy and light chains. The proteins expressed in bacteria included oligohistidine-tagged fragments of human ubiquitous kinesin heavy chain, spanning most of the stalk and all of the tail domain (amino acids 555-963); and untagged, essentially full-length human kinesin light chain (4-569) along with N-terminal (4-363)and C-terminal (364-569) light chain fragments. Heavy chain fragments were attached to Ni2+-charged beads and incubated with untagged light chain fragments. Analysis of eluted complexes by SDS-PAGE and immunoblotting mapped the light chain-binding site in heavy chain to amino acids 771-813, a region close to the C-terminal end of the heavy chain stalk domain. In addition, only the full-length and N-terminal kinesin light chain fragments bound to this heavy chain region. Within this heavy chain region are four highly conserved contiguous heptad repeats (775-802) which are predicted to form a tight α- helical coiled-coil interaction with the heptad repeat-containing N-terminus of the light chain, in particular region 106-152 of human light chain. This predicted hydrophobic, α-helical coiled-coil interaction is supported by both circular dichroism spectroscopy of the recombinant kinesin heavy chain fragment 771-963, which displays an α-helical content of 70%, and the resistance of the heavy/light chain interdiction to high salt (0.5 M).\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"fox-kowalski-king-mackay-crossley-keyresiduescharacteristicofgatanfingersarerecognizedbyfog-1998\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Key residues characteristic of GATA N-fingers are recognized by FOG.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Fox,  A.; Kowalski,  K.; King,  G.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 273(50): 33595-33603.  1998.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.273.50.33595\"\n     onclick=\"log_download('fox-kowalski-king-mackay-crossley-keyresiduescharacteristicofgatanfingersarerecognizedbyfog-1998', 'http://doi.org/10.1074/jbc.273.50.33595', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/fox-kowalski-king-mackay-crossley-keyresiduescharacteristicofgatanfingersarerecognizedbyfog-1998\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('fox-kowalski-king-mackay-crossley-keyresiduescharacteristicofgatanfingersarerecognizedbyfog-1998')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Key residues characteristic of GATA N-fingers are recognized by FOG},\n type = {article},\n year = {1998},\n pages = {33595-33603},\n volume = {273},\n id = {e9e15c4c-0ebf-3abb-b74a-16640859e1ae},\n created = {2023-01-10T01:46:48.015Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:48.015Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Protein-protein interactions play significant roles in the control of gene expression. These interactions often occur between small, discrete domains within different transcription factors. In particular, zinc fingers, usually regarded as DNA-binding domains, are now also known to be involved in mediating contacts between proteins. We have investigated the interaction between the erythroid transcription factor GATA-1 and its partner, the 9 zinc finger protein, FOG (Friend Of GATA). We demonstrate that this interaction represents a genuine finger-finger contact, which is dependent on zinc- coordinating residues within each protein. We map the contact domains to the core of the N-terminal zinc finger of GATA-1 and the 6th zinc finger of FOG. Using a scanning substitution strategy we identify key residues within the GATA-1 N-finger which are required for FOG binding. These residues are conserved in the N-fingers of all GATA proteins known to bind FOG, but are not found in the respective C-fingers. This observation may, therefore, account for the particular specificity of FOG for N-fingers. Interestingly, the key N-finger residues are seen to form a contiguous surface, when mapped onto the structure of the N-finger of GATA-1.},\n bibtype = {article},\n author = {Fox, A.H. and Kowalski, K. and King, G.F. and Mackay, J.P. and Crossley, M.},\n doi = {10.1074/jbc.273.50.33595},\n journal = {Journal of Biological Chemistry},\n number = {50}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Protein-protein interactions play significant roles in the control of gene expression. These interactions often occur between small, discrete domains within different transcription factors. In particular, zinc fingers, usually regarded as DNA-binding domains, are now also known to be involved in mediating contacts between proteins. We have investigated the interaction between the erythroid transcription factor GATA-1 and its partner, the 9 zinc finger protein, FOG (Friend Of GATA). We demonstrate that this interaction represents a genuine finger-finger contact, which is dependent on zinc- coordinating residues within each protein. We map the contact domains to the core of the N-terminal zinc finger of GATA-1 and the 6th zinc finger of FOG. Using a scanning substitution strategy we identify key residues within the GATA-1 N-finger which are required for FOG binding. These residues are conserved in the N-fingers of all GATA proteins known to bind FOG, but are not found in the respective C-fingers. This observation may, therefore, account for the particular specificity of FOG for N-fingers. Interestingly, the key N-finger residues are seen to form a contiguous surface, when mapped onto the structure of the N-finger of GATA-1.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-crossley-zincfingersarestickingtogether-1998\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Zinc fingers are sticking together.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Trends in Biochemical Sciences</i>, 23(1): 1-4.  1998.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0968-0004(97)01168-7\"\n     onclick=\"log_download('mackay-crossley-zincfingersarestickingtogether-1998', 'http://doi.org/10.1016/S0968-0004(97)01168-7', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-crossley-zincfingersarestickingtogether-1998\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-crossley-zincfingersarestickingtogether-1998')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Zinc fingers are sticking together},\n type = {article},\n year = {1998},\n pages = {1-4},\n volume = {23},\n id = {76060f13-2fda-31b9-b159-68b649adeb20},\n created = {2023-01-10T01:46:48.915Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:48.915Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n bibtype = {article},\n author = {Mackay, J.P. and Crossley, M.},\n doi = {10.1016/S0968-0004(97)01168-7},\n journal = {Trends in Biochemical Sciences},\n number = {1}\n}</pre>\n</div>\n\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"harding-mokdsi-mackay-prodigalidad-lucas-interactionsoftheantitumoragentmolybdocenedichloridewitholigonucleotides-1998\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Interactions of the Antitumor Agent Molybdocene Dichloride with Oligonucleotides.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Harding,  M.; Mokdsi,  G.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Prodigalidad,  M.;  and Lucas,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Inorganic Chemistry</i>, 37(10): 2432-2437.  1998.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/ic971205k\"\n     onclick=\"log_download('harding-mokdsi-mackay-prodigalidad-lucas-interactionsoftheantitumoragentmolybdocenedichloridewitholigonucleotides-1998', 'http://doi.org/10.1021/ic971205k', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/harding-mokdsi-mackay-prodigalidad-lucas-interactionsoftheantitumoragentmolybdocenedichloridewitholigonucleotides-1998\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('harding-mokdsi-mackay-prodigalidad-lucas-interactionsoftheantitumoragentmolybdocenedichloridewitholigonucleotides-1998')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Interactions of the Antitumor Agent Molybdocene Dichloride with Oligonucleotides},\n type = {article},\n year = {1998},\n pages = {2432-2437},\n volume = {37},\n id = {3622d446-6dc4-3dca-890e-5982c68ebdd0},\n created = {2023-01-10T01:46:49.823Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:49.823Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The interactions between the antitumor-active metallocene molybdocene dichloride (Cp2MoCl2) and four oligonucleotides have been studied by 1H and 31P NMR spectroscopy. In 50 mM salt solutions of molybdocene dichloride, hydrolysis of the halide ligands occurs to give a solution with pD 2, containing a species in which both Cp rings remain metal bound for 24 h. At pD 7, however, partial hydrolysis of the Cp rings (∼30%) occurs after 24 h. Addition of an aqueous solution of molybdocene dichloride in 50 mM salt to the self-complementary sequence d(CGCATATGCG)2, maintaining the pD at 6.0-7.0, showed no evidence for the formation of a metallocene-oligonucleotide complex, and only peaks arising from hydrolysis of molybdocene dichloride were detected. A similar result was obtained in titration experiments with the single-stranded sequence d(ATGGTA) at pD 6.5-7.0. However, at pD 3.0, new signals assigned to a molybdocene-oligonuleotide complex or complexes were detected in the 1H NMR spectrum. No change was observed in the 31P NMR spectrum. The complex or complexes formed between molybdocene dichloride and d(ATGGTA) are stable for hours at pD 3.0; at higher pD, the complex is destabilized and only peaks arising from hydrolysis of molybdocene dichloride are detected. Titration experiments at low pD with the dinucleotide dCpG showed a new set of signals in the 1H NMR spectrum, tentatively assigned to formation of a complex arising due to coordination of molybdenum to guanine N7 and/or cytosine N3. At pD 7.0, these signals disappeared. The results obtained show that stable oligonucleotide adducts are not formed in 50 mM salt at pD 7.0, and hence it is highly unlikely that formation of molybdocene - DNA adducts in vivo is the primary action that is responsible for the antitumor properties of molybdocene dichloride.},\n bibtype = {article},\n author = {Harding, M.M. and Mokdsi, G. and Mackay, J.P. and Prodigalidad, M. and Lucas, S.W.},\n doi = {10.1021/ic971205k},\n journal = {Inorganic Chemistry},\n number = {10}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The interactions between the antitumor-active metallocene molybdocene dichloride (Cp2MoCl2) and four oligonucleotides have been studied by 1H and 31P NMR spectroscopy. In 50 mM salt solutions of molybdocene dichloride, hydrolysis of the halide ligands occurs to give a solution with pD 2, containing a species in which both Cp rings remain metal bound for 24 h. At pD 7, however, partial hydrolysis of the Cp rings (∼30%) occurs after 24 h. Addition of an aqueous solution of molybdocene dichloride in 50 mM salt to the self-complementary sequence d(CGCATATGCG)2, maintaining the pD at 6.0-7.0, showed no evidence for the formation of a metallocene-oligonucleotide complex, and only peaks arising from hydrolysis of molybdocene dichloride were detected. A similar result was obtained in titration experiments with the single-stranded sequence d(ATGGTA) at pD 6.5-7.0. However, at pD 3.0, new signals assigned to a molybdocene-oligonuleotide complex or complexes were detected in the 1H NMR spectrum. No change was observed in the 31P NMR spectrum. The complex or complexes formed between molybdocene dichloride and d(ATGGTA) are stable for hours at pD 3.0; at higher pD, the complex is destabilized and only peaks arising from hydrolysis of molybdocene dichloride are detected. Titration experiments at low pD with the dinucleotide dCpG showed a new set of signals in the 1H NMR spectrum, tentatively assigned to formation of a complex arising due to coordination of molybdenum to guanine N7 and/or cytosine N3. At pD 7.0, these signals disappeared. The results obtained show that stable oligonucleotide adducts are not formed in 50 mM salt at pD 7.0, and hence it is highly unlikely that formation of molybdocene - DNA adducts in vivo is the primary action that is responsible for the antitumor properties of molybdocene dichloride.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-kowalski-fox-czolij-king-crossley-involvementofthenfingerintheselfassociationofgata1-1998\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Involvement of the N-finger in the self-association of GATA-1.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Kowalski,  K.; Fox,  A.; Czolij,  R.; King,  G.;  and Crossley,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biological Chemistry</i>, 273(46): 30560-30567.  1998.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1074/jbc.273.46.30560\"\n     onclick=\"log_download('mackay-kowalski-fox-czolij-king-crossley-involvementofthenfingerintheselfassociationofgata1-1998', 'http://doi.org/10.1074/jbc.273.46.30560', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-kowalski-fox-czolij-king-crossley-involvementofthenfingerintheselfassociationofgata1-1998\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-kowalski-fox-czolij-king-crossley-involvementofthenfingerintheselfassociationofgata1-1998')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Involvement of the N-finger in the self-association of GATA-1},\n type = {article},\n year = {1998},\n pages = {30560-30567},\n volume = {273},\n id = {4c338f22-0421-34a2-92f2-d97f844ae7db},\n created = {2023-01-10T01:46:50.749Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:50.749Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Zinc fingers are recognized as small protein domains that bind to specific DNA sequences. Recently however, zinc fingers from a number of proteins, in particular the GATA family of transcription factors, have also been implicated in specific protein-protein interactions. The erythroid protein GATA-1 contains two zinc fingers: the C-finger, which is sufficient for sequence-specific DNA-binding, and the N-finger, which appears both to modulate DNA-binding and to interact with other transcription factors. We have expressed and purified the N-finger domain and investigated its involvement in the self-association of GATA-1. We demonstrate that this domain does not homodimerize but instead makes intermolecular contacts with the C-finger, suggesting that GATA dimers are maintained by reciprocal N- finger-C-finger contacts. Deletion analysis identifies a 25-residue region, C-terminal to the core N-finger domain, that is sufficient for interaction with intact GATA-1. A similar subdomain exists C-terminal to the C-finger, and we show that self-association is substantially reduced when both subdomains are disrupted by mutation. Moreover, mutations that impair GATA-1 self-association also interfere with its ability to activate transcription in transfection studies.},\n bibtype = {article},\n author = {Mackay, J.P. and Kowalski, K. and Fox, A.H. and Czolij, R. and King, G.F. and Crossley, M.},\n doi = {10.1074/jbc.273.46.30560},\n journal = {Journal of Biological Chemistry},\n number = {46}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Zinc fingers are recognized as small protein domains that bind to specific DNA sequences. Recently however, zinc fingers from a number of proteins, in particular the GATA family of transcription factors, have also been implicated in specific protein-protein interactions. The erythroid protein GATA-1 contains two zinc fingers: the C-finger, which is sufficient for sequence-specific DNA-binding, and the N-finger, which appears both to modulate DNA-binding and to interact with other transcription factors. We have expressed and purified the N-finger domain and investigated its involvement in the self-association of GATA-1. We demonstrate that this domain does not homodimerize but instead makes intermolecular contacts with the C-finger, suggesting that GATA dimers are maintained by reciprocal N- finger-C-finger contacts. Deletion analysis identifies a 25-residue region, C-terminal to the core N-finger domain, that is sufficient for interaction with intact GATA-1. A similar subdomain exists C-terminal to the C-finger, and we show that self-association is substantially reduced when both subdomains are disrupted by mutation. Moreover, mutations that impair GATA-1 self-association also interfere with its ability to activate transcription in transfection studies.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_1997\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_1997')\"\n      onkeypress=\"toggleGroup('group_1997')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>1997</span>\n      <span class=\"bibbase_group_count\">\n        \n        (8)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"harding-krippner-shelton-rodger-sanders-mackay-prakash-dnabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'harding-krippner-shelton-rodger-sanders-mackay-prakash-dnabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997', 'https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M', event);\">\n    DNA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Harding,  M., M.; Krippner,  G., Y.; Shelton,  C., J.; Rodger,  A.; Sanders,  K., J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>;  and Prakash,  A., S.\n</span>\n\n  \n\n</span>\n\n  <i>Biopolymers</i>, 42(4): 387-398. 10 1997.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M\"\n     onclick=\"log_download('harding-krippner-shelton-rodger-sanders-mackay-prakash-dnabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997', 'https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"DNA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1002/(SICI)1097-0282(19971005)42:4&lt;387::AID-BIP2&gt;3.0.CO;2-M\"\n     onclick=\"log_download('harding-krippner-shelton-rodger-sanders-mackay-prakash-dnabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997', 'http://doi.org/10.1002/(SICI)1097-0282(19971005)42:4&lt;387::AID-BIP2&gt;3.0.CO;2-M', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/harding-krippner-shelton-rodger-sanders-mackay-prakash-dnabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('harding-krippner-shelton-rodger-sanders-mackay-prakash-dnabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {DNA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II},\n type = {article},\n year = {1997},\n keywords = {CD,DNA chemical footprinting,Fotemiistine,Intercalation,Linear dichroism,RNA polymerase II,SPXX peptides,YSPTSPSY,β-tum},\n pages = {387-398},\n volume = {42},\n websites = {https://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0282(19971005)42:4%3C387::AID-BIP2%3E3.0.CO;2-M},\n month = {10},\n day = {5},\n id = {987f67ee-49d8-3803-8df7-9251cdbaf6c9},\n created = {2020-12-17T05:29:53.650Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:53.650Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Harding1997},\n source_type = {ARTICLE},\n notes = {cited By 5},\n private_publication = {false},\n abstract = {The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley &amp; Sons, Inc.},\n bibtype = {article},\n author = {Harding, Margaret M. and Krippner, Guy Y. and Shelton, Cathryn J. and Rodger, Alison and Sanders, Karen J. and Mackay, Joel P. and Prakash, Arungundrum S.},\n doi = {10.1002/(SICI)1097-0282(19971005)42:4&lt;387::AID-BIP2&gt;3.0.CO;2-M},\n journal = {Biopolymers},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley & Sons, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"https://link.springer.com/article/10.1023/A:1018339526108\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997', 'https://link.springer.com/article/10.1023/A:1018339526108', event);\">\n    Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dingley,  A., J.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Shaw,  G., L.; Hambly,  B., D.;  and King,  G., F.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biomolecular NMR</i>, 10(1): 1-8.  1997.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"https://link.springer.com/article/10.1023/A:1018339526108\"\n     onclick=\"log_download('dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997', 'https://link.springer.com/article/10.1023/A:1018339526108', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/https://doi.org/10.1023/A:1018339526108\"\n     onclick=\"log_download('dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997', 'http://doi.org/https://doi.org/10.1023/A:1018339526108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR},\n type = {article},\n year = {1997},\n keywords = {Macromolecules,Pulsed-field-gradient NMR,Self-association,Solvent suppression,Translational diffusion coefficient},\n pages = {1-8},\n volume = {10},\n websites = {https://link.springer.com/article/10.1023/A:1018339526108},\n id = {c4559b9b-27f6-3507-a99a-884b885fa4fb},\n created = {2020-12-17T05:29:57.841Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:57.841Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Dingley1997},\n source_type = {ARTICLE},\n notes = {cited By 24},\n private_publication = {false},\n abstract = {We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.},\n bibtype = {article},\n author = {Dingley, Andrew J. and Mackay, Joel P. and Shaw, Graeme L. and Hambly, Brett D. and King, Glenn F.},\n doi = {https://doi.org/10.1023/A:1018339526108},\n journal = {Journal of Biomolecular NMR},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Bains,  N.; Wilce,  J.; Heuer,  K.; Tunstall,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Bennett,  M.; Weiss,  A.;  and King,  G.\n</span>\n\n  \n\n</span>\n\n  <i>International Journal of Peptide Research and Therapeutics</i>, 4(2): 67-77.  1997.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/BF02443517\"\n     onclick=\"log_download('bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997', 'http://doi.org/10.1007/BF02443517', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides},\n type = {article},\n year = {1997},\n pages = {67-77},\n volume = {4},\n id = {e3e2a974-5c9b-3e6e-8d6b-ec39b861bc0a},\n created = {2023-01-10T01:46:51.663Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:51.663Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides. © 1997 ESCOM Science Publishers B.V.},\n bibtype = {article},\n author = {Bains, N.P.S. and Wilce, J.A. and Heuer, K.H. and Tunstall, M. and Mackay, J.P. and Bennett, M.R. and Weiss, A.S. and King, G.F.},\n doi = {10.1007/BF02443517},\n journal = {International Journal of Peptide Research and Therapeutics},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides. © 1997 ESCOM Science Publishers B.V.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dingley,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Shaw,  G.; Hambly,  B.;  and King,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biomolecular NMR</i>, 10(1): 1-8.  1997.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1023/A:1018339526108\"\n     onclick=\"log_download('dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997', 'http://doi.org/10.1023/A:1018339526108', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dingley-mackay-shaw-hambly-king-measuringmacromoleculardiffusionusingheteronuclearmultiplequantumpulsedfieldgradientnmr-1997')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR},\n type = {article},\n year = {1997},\n pages = {1-8},\n volume = {10},\n id = {53beac46-6091-3dfa-988a-801d2a1ac05f},\n created = {2023-01-10T01:46:52.607Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:52.607Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.},\n bibtype = {article},\n author = {Dingley, A.J. and Mackay, J.P. and Shaw, G.L. and Hambly, B.D. and King, G.F.},\n doi = {10.1023/A:1018339526108},\n journal = {Journal of Biomolecular NMR},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  We have previously shown that 1H pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for 15N-labelled proteins, selection of 1H-15N multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring 1H single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, 1H-15N zero-quantum coherence dephases more rapidly than 1H SQ coherence under the influence of a PFG This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"fletcher-chapman-mackay-howden-king-thestructureofversutoxinatracotoxinhv1providesinsightsintothebindingofsite3neurotoxinstothevoltagegatedsodiumchannel-1997\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The structure of versutoxin (δ-atracotoxin-Hv1) provides insights into the binding of site 3 neurotoxins to the voltage-gated sodium channel.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Fletcher,  J.; Chapman,  B.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Howden,  M.;  and King,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 5(11): 1525-1535.  1997.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0969-2126(97)00301-8\"\n     onclick=\"log_download('fletcher-chapman-mackay-howden-king-thestructureofversutoxinatracotoxinhv1providesinsightsintothebindingofsite3neurotoxinstothevoltagegatedsodiumchannel-1997', 'http://doi.org/10.1016/S0969-2126(97)00301-8', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/fletcher-chapman-mackay-howden-king-thestructureofversutoxinatracotoxinhv1providesinsightsintothebindingofsite3neurotoxinstothevoltagegatedsodiumchannel-1997\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('fletcher-chapman-mackay-howden-king-thestructureofversutoxinatracotoxinhv1providesinsightsintothebindingofsite3neurotoxinstothevoltagegatedsodiumchannel-1997')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The structure of versutoxin (δ-atracotoxin-Hv1) provides insights into the binding of site 3 neurotoxins to the voltage-gated sodium channel},\n type = {article},\n year = {1997},\n pages = {1525-1535},\n volume = {5},\n id = {c641551b-c512-3935-9b0e-32d0d876b55e},\n created = {2023-01-10T01:46:53.511Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:53.511Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background: Versutoxin (δ-ACTX-Hv1) is the major component of the venom of the Australian Blue Mountains funnel web spider, Hadronyche versuta. δ-ACTX-Hv1 produces potentially fatal neurotoxic symptoms in primates by slowing the inactivation of voltage-gated sodium channels; δ-ACTX-Hv1 is therefore a useful tool for studying sodium channel function. We have determined the three-dimensional structure of δ-ACTX-Hv1 as the first step towards understanding the molecular basis of its interaction with these channels. Results: The solution structure of δ-ACTX-Hv1, determined using NMR spectroscopy, comprises a core β region containing a triple-stranded antiparallel β sheet, a thumb-like extension protruding from the β region and a C-terminal 310 helix that is appended to the β domain by virtue of a disulphide bond. The β region contains a cystine knot motif similar to that seen in other neurotoxic polypeptides. The structure shows homology with μ-agatoxin-I, a spider toxin that also modifies the inactivation kinetics of vertebrate voltage-gated sodium channels. More surprisingly, δ-ACTX-Hv1 shows both sequence and structural homology with gurmarin, a plant polypeptide. This similarity leads us to suggest that the sweet-taste suppression elicited by gurmarin may result from an interaction with one of the downstream ion channels involved in sweet-taste transduction. Conclusions: δ-ACTX-Hv1 shows no structural homology with either sea anemone or α-scorpion toxins, both of which also modify the inactivation kinetics of voltage-gated sodium channels by interacting with channel recognition site 3. However, we have shown that δ-ACTX-Hv1 contains charged residues that are topologically related to those implicated in the binding of sea anemone and α-scorpion toxins to mammalian voltage-gated sodium channels, suggesting similarities in their mode of interaction with these channels.},\n bibtype = {article},\n author = {Fletcher, J.I. and Chapman, B.E. and Mackay, J.P. and Howden, M.E.H. and King, G.F.},\n doi = {10.1016/S0969-2126(97)00301-8},\n journal = {Structure},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background: Versutoxin (δ-ACTX-Hv1) is the major component of the venom of the Australian Blue Mountains funnel web spider, Hadronyche versuta. δ-ACTX-Hv1 produces potentially fatal neurotoxic symptoms in primates by slowing the inactivation of voltage-gated sodium channels; δ-ACTX-Hv1 is therefore a useful tool for studying sodium channel function. We have determined the three-dimensional structure of δ-ACTX-Hv1 as the first step towards understanding the molecular basis of its interaction with these channels. Results: The solution structure of δ-ACTX-Hv1, determined using NMR spectroscopy, comprises a core β region containing a triple-stranded antiparallel β sheet, a thumb-like extension protruding from the β region and a C-terminal 310 helix that is appended to the β domain by virtue of a disulphide bond. The β region contains a cystine knot motif similar to that seen in other neurotoxic polypeptides. The structure shows homology with μ-agatoxin-I, a spider toxin that also modifies the inactivation kinetics of vertebrate voltage-gated sodium channels. More surprisingly, δ-ACTX-Hv1 shows both sequence and structural homology with gurmarin, a plant polypeptide. This similarity leads us to suggest that the sweet-taste suppression elicited by gurmarin may result from an interaction with one of the downstream ion channels involved in sweet-taste transduction. Conclusions: δ-ACTX-Hv1 shows no structural homology with either sea anemone or α-scorpion toxins, both of which also modify the inactivation kinetics of voltage-gated sodium channels by interacting with channel recognition site 3. However, we have shown that δ-ACTX-Hv1 contains charged residues that are topologically related to those implicated in the binding of sea anemone and α-scorpion toxins to mammalian voltage-gated sodium channels, suggesting similarities in their mode of interaction with these channels.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Bains,  N.; Wilce,  J.; Heuer,  K.; Tunstall,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Bennett,  M.; Weiss,  A.;  and King,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Letters in Peptide Science</i>, 4(2): 67-77.  1997.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/BF02443517\"\n     onclick=\"log_download('bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997', 'http://doi.org/10.1007/BF02443517', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('bains-wilce-heuer-tunstall-mackay-bennett-weiss-king-zippinguptranscriptionfactorsrationaldesignofantijunandantifospeptides-1997')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Zipping up transcription factors: Rational design of anti-Jun and anti-Fos peptides},\n type = {article},\n year = {1997},\n pages = {67-77},\n volume = {4},\n id = {424daf6d-00c1-3bcf-ad60-9abdb95c2e4b},\n created = {2023-01-10T01:46:54.417Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:54.417Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides.},\n bibtype = {article},\n author = {Bains, N.P.S. and Wilce, J.A. and Heuer, K.H. and Tunstall, M. and Mackay, J.P. and Bennett, M.R. and Weiss, A.S. and King, G.F.},\n doi = {10.1007/BF02443517},\n journal = {Letters in Peptide Science},\n number = {2}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Various members of the bZip and bHLH-Zip families of eukaryotic transcription factors, including Jun, Fos, and Myc, have been identified as oncoproteins; mutation or deregulated expression of these proteins leads to certain types of cancer. These proteins can only bind to their cognate DNA enhancer sites following homodimerization, or heterodimerization with another family member, via their leucine zipper domain. Thus, a novel anticancer strategy would be to inhibit dimerization of these proteins, thereby blocking their DNA binding and transactivation functions. In this paper we show that it is possible to rationally design leucine zipper peptides that bind with high affinity to the leucine zipper dimerization domains of c-Jun and c-Fos, thus preventing the formation of functional c-Jun homodimers and c-Jun:c-Fos heterodimers; we refer to such peptides as superzippers (SZs). In vivo, c-Jun:SZ and c-Fos:SZ heterodimers should be nonfunctional as they lack one of the two basic domains that are essential for DNA binding. While the transport of a peptidic agent into cells often poses a severe obstacle to its therapeutic use, we show that a 46-residue leucine zipper peptide can be transported into HeLa cells by coupling it to a 17-residue carrier peptide from the Antennapedia homeodomain, thus paving the way for detailed studies of the therapeutic potential of superzipper peptides.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"harding-krippner-shelton-rodger-sanders-mackay-prakash-dmabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  DMA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Harding,  M.; Krippner,  G.; Shelton,  C.; Rodger,  A.; Sanders,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Prakash,  A.\n</span>\n\n  \n\n</span>\n\n  <i>Biopolymers - Nucleic Acid Sciences Section</i>, 42(4): 387-398.  1997.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1002/(SICI)1097-0282(19971005)42:4&lt;387::AID-BIP2&gt;3.0.CO;2-M\"\n     onclick=\"log_download('harding-krippner-shelton-rodger-sanders-mackay-prakash-dmabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997', 'http://doi.org/10.1002/(SICI)1097-0282(19971005)42:4&lt;387::AID-BIP2&gt;3.0.CO;2-M', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/harding-krippner-shelton-rodger-sanders-mackay-prakash-dmabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('harding-krippner-shelton-rodger-sanders-mackay-prakash-dmabindingstudiesofxsptspszderivativesoftheintercalatingheptadrepeatofrnapolymeraseii-1997')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {DMA-binding studies of XSPTSPSZ, derivatives of the intercalating heptad repeat of RNA polymerase II},\n type = {article},\n year = {1997},\n pages = {387-398},\n volume = {42},\n id = {964a530f-b34f-3e6c-917f-3ea08bb03126},\n created = {2023-01-10T01:46:55.338Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:55.338Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley &amp; Sons, Inc.},\n bibtype = {article},\n author = {Harding, M.M. and Krippner, G.Y. and Shelton, C.J. and Rodger, A. and Sanders, K.J. and Mackay, J.P. and Prakash, A.S.},\n doi = {10.1002/(SICI)1097-0282(19971005)42:4&lt;387::AID-BIP2&gt;3.0.CO;2-M},\n journal = {Biopolymers - Nucleic Acid Sciences Section},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The synthesis, solution conformation, and interaction with DNA of three 8-residiie peptides structurally related to the heptad repeat unit found at the C-termimis ofRNA polymerase II are reported. Peptides QQ, XQ, and PQ are derived from the parent sequence YSPTSPSY (peptide YY), which was reported to bind to DNA by bisintercalation [M. Suzuki (1990) Nature, Vol. 344, pp. 562-565], and contain either a 2-qiiinolyl (Q), 2-quinoxolyl (X), or 5phenanthrolyl (P) group in place of the aromatic side chains of the N- and C-tenninal tyrosine residues present in the parent sequence. The combined results of linear dichroism and induced CD measurements of peptides QQ, XQ, and PQ with calf thymus DNA are consistent with weak binding of the peptides to DNA in a preferred orientation in which the chromophores are intercalated. Small increases in the melting temperatures ofpoly[d(A-T)2] are also consistent with the peptides interacting with DNA. While enzymatic footprinting with DNase I showed no protection from cleavage by the enzyme, chemical footprinting with fotemustine showed that the peptides modify the reactivity of the major groove, presumably via minor groove binding. Peptide QQ inhibited fotemustine alkylation significantly more than either XQ or PQ, and slightly more than YY. In aqueous solution, nmr experiments on QQ, XQ, and PQ show a significant population of a conformation in which Ser2-Pro3-Thr4-Ser5 form both type I and type II β-tum conformations in equilibrium with open chain conformations. Nuclear magnetic resonance titration experiments ofPQ with (GCGTACGC)2 showed small changes in chemical shifts, consistent with the formation of a weak nonspecific complex. Analogous experiments, using peptides QQ and XQ with (GCGTACGC)2, and peplide YY with (CGTACG)2, showed no evidence for the interaction of the peptides with these oligonucleotides. These results show that peptides of general structure XSPTSPSZ are weak nonspecific DNA binders that differ significantly from previously characterized S(T)PXX DNA-binding motifs that are generally AT-selective minor groove binders. ©1997 John Wiley & Sons, Inc.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"proudfoot-mackay-vagg-vickery-williams-karuso-cisrurrpicchxnmeinf2infphensup2supshowsminorgrooveatselectivitywitholigonucleotides-1997\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Δ-cis-α-[Ru(RR-picchxnMe<inf>2</inf>)(phen)]<sup>2+</sup> shows minor groove AT selectivity with oligonucleotides.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Proudfoot,  E.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Vagg,  R.; Vickery,  K.; Williams,  P.;  and Karuso,  P.\n</span>\n\n  \n\n</span>\n\n  <i>Chemical Communications</i>, (17): 1623-1624.  1997.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1039/a704001f\"\n     onclick=\"log_download('proudfoot-mackay-vagg-vickery-williams-karuso-cisrurrpicchxnmeinf2infphensup2supshowsminorgrooveatselectivitywitholigonucleotides-1997', 'http://doi.org/10.1039/a704001f', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/proudfoot-mackay-vagg-vickery-williams-karuso-cisrurrpicchxnmeinf2infphensup2supshowsminorgrooveatselectivitywitholigonucleotides-1997\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('proudfoot-mackay-vagg-vickery-williams-karuso-cisrurrpicchxnmeinf2infphensup2supshowsminorgrooveatselectivitywitholigonucleotides-1997')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Δ-cis-α-[Ru(RR-picchxnMe&lt;inf&gt;2&lt;/inf&gt;)(phen)]&lt;sup&gt;2+&lt;/sup&gt; shows minor groove AT selectivity with oligonucleotides},\n type = {article},\n year = {1997},\n pages = {1623-1624},\n id = {9b3e2289-c499-39c6-9a25-43e57803b26c},\n created = {2023-01-10T01:46:56.245Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:56.245Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {NMR studies show that the ternary octahedral Δ-cis-α-[Ru(RR-picchxnMe2)(phen)]2+ cation binds with AT selectivity in the minor groove of [d(CGCGATCGCG)2] and [d(ATATCGATAT)2] duplexes through a non-intercalative interaction.},\n bibtype = {article},\n author = {Proudfoot, E.M. and Mackay, J.P. and Vagg, R.S. and Vickery, K.A. and Williams, P.A. and Karuso, P.},\n doi = {10.1039/a704001f},\n journal = {Chemical Communications},\n number = {17}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  NMR studies show that the ternary octahedral Δ-cis-α-[Ru(RR-picchxnMe2)(phen)]2+ cation binds with AT selectivity in the minor groove of [d(CGCGATCGCG)2] and [d(ATATCGATAT)2] duplexes through a non-intercalative interaction.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_1996\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_1996')\"\n      onkeypress=\"toggleGroup('group_1996')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>1996</span>\n      <span class=\"bibbase_group_count\">\n        \n        (3)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"mackay-shelton-harding-assignmentofthesup1suphnmrspectrumandsolutionconformationoftheantitumourantibioticditrisarubicinb-1996\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Assignment of the <sup>1</sup>H NMR spectrum and solution conformation of the antitumour antibiotic ditrisarubicin B.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Shelton,  C.;  and Harding,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Tetrahedron</i>, 52(15): 5617-5624.  1996.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/0040-4020(96)00198-6\"\n     onclick=\"log_download('mackay-shelton-harding-assignmentofthesup1suphnmrspectrumandsolutionconformationoftheantitumourantibioticditrisarubicinb-1996', 'http://doi.org/10.1016/0040-4020(96)00198-6', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-shelton-harding-assignmentofthesup1suphnmrspectrumandsolutionconformationoftheantitumourantibioticditrisarubicinb-1996\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-shelton-harding-assignmentofthesup1suphnmrspectrumandsolutionconformationoftheantitumourantibioticditrisarubicinb-1996')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Assignment of the &lt;sup&gt;1&lt;/sup&gt;H NMR spectrum and solution conformation of the antitumour antibiotic ditrisarubicin B},\n type = {article},\n year = {1996},\n pages = {5617-5624},\n volume = {52},\n id = {47129b2c-d23a-3485-8fba-32923cad98a3},\n created = {2023-01-10T01:46:57.152Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:57.152Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Complete assignment of the 1H NMR spectrum of ditrisarubicin B, a member of the anthracycline antitumour antibiotics, in acetonitrile is reported. The two trisaccharide chains are highly structured and their conformation supports their role as preorganised DNA minor groove binders which bind to DNA on intercalation of the tetracyclic chromophore.},\n bibtype = {article},\n author = {Mackay, J.P. and Shelton, C.J. and Harding, M.M.},\n doi = {10.1016/0040-4020(96)00198-6},\n journal = {Tetrahedron},\n number = {15}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Complete assignment of the 1H NMR spectrum of ditrisarubicin B, a member of the anthracycline antitumour antibiotics, in acetonitrile is reported. The two trisaccharide chains are highly structured and their conformation supports their role as preorganised DNA minor groove binders which bind to DNA on intercalation of the tetracyclic chromophore.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-shaw-king-backbonedynamicsofthecjunleucinezippersup15supnnmrrelaxationstudies-1996\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Backbone dynamics of the c-Jun leucine zipper: <sup>15</sup>N NMR relaxation studies.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Shaw,  G.;  and King,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 35(15): 4867-4877.  1996.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi952761y\"\n     onclick=\"log_download('mackay-shaw-king-backbonedynamicsofthecjunleucinezippersup15supnnmrrelaxationstudies-1996', 'http://doi.org/10.1021/bi952761y', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-shaw-king-backbonedynamicsofthecjunleucinezippersup15supnnmrrelaxationstudies-1996\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-shaw-king-backbonedynamicsofthecjunleucinezippersup15supnnmrrelaxationstudies-1996')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Backbone dynamics of the c-Jun leucine zipper: &lt;sup&gt;15&lt;/sup&gt;N NMR relaxation studies},\n type = {article},\n year = {1996},\n pages = {4867-4877},\n volume = {35},\n id = {362dbcd0-e82e-357a-a5d5-97ec3d2ac62e},\n created = {2023-01-10T01:46:58.102Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:58.102Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {The backbone dynamics of the coiled-coil leucine zipper domain of c-Jun have been studied using proton-detected two-dimensional 1H-15N NMR spectroscopy. Longitudinal (T1) and transverse (T2) 15N relaxation times, together with 1H15N NOEs, were measured and analyzed by considering the protein to approximate a prolate ellipsoid. An analysis of the T1/T2 ratios for residues in the well-structured section of the protein showed that a model for the spectral density function in which the protein is considered to reorient anisotropically fitted the data significantly better than an isotropic model. Order parameters (S2) in the range 0.7-0.9 were observed for most residues, with lower values near the C-terminus, consistent with fraying of the two helices comprising the coiled-coil. Because nearly all of the N-H vectors have small angles to the long axis of the molecule, there was some uncertainty in the value of the rotational diffusion coefficient Dpar, which describes rotation about the long axis. Thus, an alternative method was examined for its ability to provide independent estimates of Dpar and Dperp (the diffusion coefficient describing rotation about axes perpendicular to the long axis); the translational diffusion coefficient (Dt) of the protein was measured, and hydrodynamic calculations were used to predict Dpar and Dperp. However, the derived rotational diffusion coefficients proved to be very dependent on the hydrodynamic model used to relate Dt to Dpar and Dperp, and consequently the values obtained from the T1/T2 analysis were used in the order-parameter analysis. Although it has previously been reported that the side chain of a polar residue at the dimer interface, Asn22, undergoes a conformational exchange process and destabilizes the dimer, no evidence of increased backbone mobility in this region was detected, suggesting that this process is confined to the Asn side chain.},\n bibtype = {article},\n author = {Mackay, J.P. and Shaw, G.L. and King, G.F.},\n doi = {10.1021/bi952761y},\n journal = {Biochemistry},\n number = {15}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  The backbone dynamics of the coiled-coil leucine zipper domain of c-Jun have been studied using proton-detected two-dimensional 1H-15N NMR spectroscopy. Longitudinal (T1) and transverse (T2) 15N relaxation times, together with 1H15N NOEs, were measured and analyzed by considering the protein to approximate a prolate ellipsoid. An analysis of the T1/T2 ratios for residues in the well-structured section of the protein showed that a model for the spectral density function in which the protein is considered to reorient anisotropically fitted the data significantly better than an isotropic model. Order parameters (S2) in the range 0.7-0.9 were observed for most residues, with lower values near the C-terminus, consistent with fraying of the two helices comprising the coiled-coil. Because nearly all of the N-H vectors have small angles to the long axis of the molecule, there was some uncertainty in the value of the rotational diffusion coefficient Dpar, which describes rotation about the long axis. Thus, an alternative method was examined for its ability to provide independent estimates of Dpar and Dperp (the diffusion coefficient describing rotation about axes perpendicular to the long axis); the translational diffusion coefficient (Dt) of the protein was measured, and hydrodynamic calculations were used to predict Dpar and Dperp. However, the derived rotational diffusion coefficients proved to be very dependent on the hydrodynamic model used to relate Dt to Dpar and Dperp, and consequently the values obtained from the T1/T2 analysis were used in the order-parameter analysis. Although it has previously been reported that the side chain of a polar residue at the dimer interface, Asn22, undergoes a conformational exchange process and destabilizes the dimer, no evidence of increased backbone mobility in this region was detected, suggesting that this process is confined to the Asn side chain.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"heuer-mackay-podzebenko-bains-weiss-king-easterbrooksmith-developmentofasensitivepeptidebasedimmunoassayapplicationtodetectionofthejunandfosoncoproteins-1996\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Development of a sensitive peptide-based immunoassay: Application to detection of the Jun and Fos oncoproteins.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Heuer,  K.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Podzebenko,  P.; Bains,  N.; Weiss,  A.; King,  G.;  and Easterbrook-Smith,  S.\n</span>\n\n  \n\n</span>\n\n  <i>Biochemistry</i>, 35(28): 9069-9075.  1996.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/bi952817o\"\n     onclick=\"log_download('heuer-mackay-podzebenko-bains-weiss-king-easterbrooksmith-developmentofasensitivepeptidebasedimmunoassayapplicationtodetectionofthejunandfosoncoproteins-1996', 'http://doi.org/10.1021/bi952817o', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/heuer-mackay-podzebenko-bains-weiss-king-easterbrooksmith-developmentofasensitivepeptidebasedimmunoassayapplicationtodetectionofthejunandfosoncoproteins-1996\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('heuer-mackay-podzebenko-bains-weiss-king-easterbrooksmith-developmentofasensitivepeptidebasedimmunoassayapplicationtodetectionofthejunandfosoncoproteins-1996')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Development of a sensitive peptide-based immunoassay: Application to detection of the Jun and Fos oncoproteins},\n type = {article},\n year = {1996},\n pages = {9069-9075},\n volume = {35},\n id = {3ec35428-2577-35f5-8d93-7aae09b3b3bc},\n created = {2023-01-10T01:46:59.013Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:46:59.013Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {c-Jun and c-Fos belong to the bZIP class of transcriptional activator proteins, many of which have been implicated in the neoplastic transformation of cells. We are interested in engineering dominant-negative leucine zipper (LZ) peptides as a means of sequestering these proteins in vivo in order to suppress their transcriptional regulatory activity. Toward this end, we have developed a novel immunoassay for measuring the dimerization affinities of dimeric Jun and Fos complexes. This peptide-based ELISA relies on the fact that Jun and Fos preferentially form heterodimers via their leucine zipper domains. Recombinant Jun leucine zipper peptides (either native JunLZ or a V36→E point mutant) were labeled with biotin and specifically bound through a leucine zipper interaction to a FosLZ-glutathione S-transferase fusion protein adsorbed onto the wells of an ELISA tray. Jun:Fos complexes were subsequently detected using a recently developed streptavidin-based amplification system known as enzyme complex amplification [Wilson, M. R., and Easterbrook-Smith, S. B. (1993) Anal. Biochem. 209, 183-187]. This ELISA system can detect subnanomolar concentrations of Jun and Fos, thus allowing determination of the dissociation constants for complex formation. The dissociation constant for formation of the native JunLZ:FosLZ heterodimer at 37 °C was determined to be 0.99 ± 0.30 nM, while that for JunLZ(V36E):FosLZ heterodimer was 0.90 ± 0.13 μM. These results demonstrate that the novel peptide-based ELISA described herein is simple and sensitive and can be used to rapidly screen for potential dominant-negative leucine zipper peptides.},\n bibtype = {article},\n author = {Heuer, K.H. and Mackay, J.P. and Podzebenko, P. and Bains, N.P.S. and Weiss, A.S. and King, G.F. and Easterbrook-Smith, S.B.},\n doi = {10.1021/bi952817o},\n journal = {Biochemistry},\n number = {28}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  c-Jun and c-Fos belong to the bZIP class of transcriptional activator proteins, many of which have been implicated in the neoplastic transformation of cells. We are interested in engineering dominant-negative leucine zipper (LZ) peptides as a means of sequestering these proteins in vivo in order to suppress their transcriptional regulatory activity. Toward this end, we have developed a novel immunoassay for measuring the dimerization affinities of dimeric Jun and Fos complexes. This peptide-based ELISA relies on the fact that Jun and Fos preferentially form heterodimers via their leucine zipper domains. Recombinant Jun leucine zipper peptides (either native JunLZ or a V36→E point mutant) were labeled with biotin and specifically bound through a leucine zipper interaction to a FosLZ-glutathione S-transferase fusion protein adsorbed onto the wells of an ELISA tray. Jun:Fos complexes were subsequently detected using a recently developed streptavidin-based amplification system known as enzyme complex amplification [Wilson, M. R., and Easterbrook-Smith, S. B. (1993) Anal. Biochem. 209, 183-187]. This ELISA system can detect subnanomolar concentrations of Jun and Fos, thus allowing determination of the dissociation constants for complex formation. The dissociation constant for formation of the native JunLZ:FosLZ heterodimer at 37 °C was determined to be 0.99 ± 0.30 nM, while that for JunLZ(V36E):FosLZ heterodimer was 0.90 ± 0.13 μM. These results demonstrate that the novel peptide-based ELISA described herein is simple and sensitive and can be used to rapidly screen for potential dominant-negative leucine zipper peptides.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_1995\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_1995')\"\n      onkeypress=\"toggleGroup('group_1995')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>1995</span>\n      <span class=\"bibbase_group_count\">\n        \n        (1)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"dingley-mackay-chapman-morris-kuchel-hambly-king-measuringproteinselfassociationusingpulsedfieldgradientnmrspectroscopyapplicationtomyosinlightchain2-1995\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Measuring protein self-association using pulsed-field-gradient NMR spectroscopy: Application to myosin light chain 2.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Dingley,  A.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Chapman,  B.; Morris,  M.; Kuchel,  P.; Hambly,  B.;  and King,  G.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of Biomolecular NMR</i>, 6(3): 321-328.  1995.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1007/BF00197813\"\n     onclick=\"log_download('dingley-mackay-chapman-morris-kuchel-hambly-king-measuringproteinselfassociationusingpulsedfieldgradientnmrspectroscopyapplicationtomyosinlightchain2-1995', 'http://doi.org/10.1007/BF00197813', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/dingley-mackay-chapman-morris-kuchel-hambly-king-measuringproteinselfassociationusingpulsedfieldgradientnmrspectroscopyapplicationtomyosinlightchain2-1995\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('dingley-mackay-chapman-morris-kuchel-hambly-king-measuringproteinselfassociationusingpulsedfieldgradientnmrspectroscopyapplicationtomyosinlightchain2-1995')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Measuring protein self-association using pulsed-field-gradient NMR spectroscopy: Application to myosin light chain 2},\n type = {article},\n year = {1995},\n pages = {321-328},\n volume = {6},\n id = {a162714d-04d2-3f71-8080-eeb26343a53a},\n created = {2023-01-10T01:47:00.198Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:47:00.198Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {At the millimolar concentrations required for structural studies, NMR spectra of the calcium-binding protein myosin light chain 2 (MLC2) showed resonance line widths indicative of extensive self-association. Pulsed-field-gradient (PFG) NMR spectroscopy was used to examine whether MLC2 aggregation could be prevented by the zwitterionic bile salt derivative 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). PFG NMR measurements indicated that CHAPS was capable of preventing MLC2 self-association, but only at concentrations well above the critical micelle concentration of ∼7.5 mM. CHAPS was most effective at a concentration of 22.5 mM, where the apparent molecular mass of MLC2 correponded to a protein monomer plus seven molecules of bound detergent. The resolution and sensitivity of 2D 15N-1H HSQC spectra of MLC2 were markedly improved by the addition of 25 mM CHAPS, consistent with a reduction in aggregation following addition of the detergent. The average amide nitrogen T2 value for MLC2 increased from ∼30 ms in the absence of CHAPS to ∼56 ms in the presence of 25 mM CHAPS. The results of this study lead us to propose that PFG NMR spectroscopy can be used as a facile alternative to conventional techniques such as analytical ultracentrifugation for examining the self-association of biological macromolecules. © 1995 ESCOM Science Publishers B.V.},\n bibtype = {article},\n author = {Dingley, A.J. and Mackay, J.P. and Chapman, B.E. and Morris, M.B. and Kuchel, P.W. and Hambly, B.D. and King, G.F.},\n doi = {10.1007/BF00197813},\n journal = {Journal of Biomolecular NMR},\n number = {3}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  At the millimolar concentrations required for structural studies, NMR spectra of the calcium-binding protein myosin light chain 2 (MLC2) showed resonance line widths indicative of extensive self-association. Pulsed-field-gradient (PFG) NMR spectroscopy was used to examine whether MLC2 aggregation could be prevented by the zwitterionic bile salt derivative 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). PFG NMR measurements indicated that CHAPS was capable of preventing MLC2 self-association, but only at concentrations well above the critical micelle concentration of ∼7.5 mM. CHAPS was most effective at a concentration of 22.5 mM, where the apparent molecular mass of MLC2 correponded to a protein monomer plus seven molecules of bound detergent. The resolution and sensitivity of 2D 15N-1H HSQC spectra of MLC2 were markedly improved by the addition of 25 mM CHAPS, consistent with a reduction in aggregation following addition of the detergent. The average amide nitrogen T2 value for MLC2 increased from ∼30 ms in the absence of CHAPS to ∼56 ms in the presence of 25 mM CHAPS. The results of this study lead us to propose that PFG NMR spectroscopy can be used as a facile alternative to conventional techniques such as analytical ultracentrifugation for examining the self-association of biological macromolecules. © 1995 ESCOM Science Publishers B.V.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_1994\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_1994')\"\n      onkeypress=\"toggleGroup('group_1994')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>1994</span>\n      <span class=\"bibbase_group_count\">\n        \n        (4)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"williams-searle-groves-mackay-westwell-beauregard-cristofaro-functionalrolesofnaturalproductstheinvolvementofextendedarraysofweakinteractionsincooperativebindingphenomena-1994\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Functional roles of natural products: The involvement of extended arrays of weak interactions in cooperative binding phenomena.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Williams,  D.; Searle,  M.; Groves,  P.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Westwell,  M.; Beauregard,  D.;  and Cristofaro,  M.\n</span>\n\n  \n\n</span>\n\n  <i>Pure and Applied Chemistry</i>, 66(10-11): 1975-1982.  1994.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1351/pac199466101975\"\n     onclick=\"log_download('williams-searle-groves-mackay-westwell-beauregard-cristofaro-functionalrolesofnaturalproductstheinvolvementofextendedarraysofweakinteractionsincooperativebindingphenomena-1994', 'http://doi.org/10.1351/pac199466101975', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/williams-searle-groves-mackay-westwell-beauregard-cristofaro-functionalrolesofnaturalproductstheinvolvementofextendedarraysofweakinteractionsincooperativebindingphenomena-1994\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('williams-searle-groves-mackay-westwell-beauregard-cristofaro-functionalrolesofnaturalproductstheinvolvementofextendedarraysofweakinteractionsincooperativebindingphenomena-1994')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Functional roles of natural products: The involvement of extended arrays of weak interactions in cooperative binding phenomena},\n type = {article},\n year = {1994},\n pages = {1975-1982},\n volume = {66},\n id = {c10c940c-9f14-3cf0-888e-39c896da615c},\n created = {2023-01-10T01:47:01.128Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:47:01.128Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {A factorisation of the free energy of binding into various “costs” and “benefits” has provided the basis for a semi-quantitation of some weak interactions in solution. It has become clear that the entropie cost of motional restriction on binding (A + B → A.B) increases with increasing exothermicity of the association; this exothermicity must of course reflect not only interactions at the interface between A and B, but also the change in bonding throughout B (if B represents the receptor). We illustrate cooperativity and anti-cooperativity by reference to effects of ligand binding (as models of classical agonists and antagonists) on the dimerisation of vancomycin-group antibiotics. Since dimerization of receptors (promoted by ligand binding) is a common theme in biological signalling, it must presumably have an advantage in natural selection. We suggest that concurrent demands of ligand binding and receptor dimerization may permit a more specific control of those ligand structures which can cause signal transmission. In this way, specificity in biological signalling might be aided. © 1994 IUPAC},\n bibtype = {article},\n author = {Williams, D.H. and Searle, M.S. and Groves, P. and Mackay, J.P. and Westwell, M.S. and Beauregard, D.A. and Cristofaro, M.F.},\n doi = {10.1351/pac199466101975},\n journal = {Pure and Applied Chemistry},\n number = {10-11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A factorisation of the free energy of binding into various “costs” and “benefits” has provided the basis for a semi-quantitation of some weak interactions in solution. It has become clear that the entropie cost of motional restriction on binding (A + B → A.B) increases with increasing exothermicity of the association; this exothermicity must of course reflect not only interactions at the interface between A and B, but also the change in bonding throughout B (if B represents the receptor). We illustrate cooperativity and anti-cooperativity by reference to effects of ligand binding (as models of classical agonists and antagonists) on the dimerisation of vancomycin-group antibiotics. Since dimerization of receptors (promoted by ligand binding) is a common theme in biological signalling, it must presumably have an advantage in natural selection. We suggest that concurrent demands of ligand binding and receptor dimerization may permit a more specific control of those ligand structures which can cause signal transmission. In this way, specificity in biological signalling might be aided. © 1994 IUPAC\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-gerhard-beauregard-maplestone-williams-dissectionofthecontributionstowarddimerizationofglycopeptideantibiotics-1994\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Dissection of the Contributions toward Dimerization of Glycopeptide Antibiotics.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Gerhard,  U.; Beauregard,  D.; Maplestone,  R.;  and Williams,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of the American Chemical Society</i>, 116(11): 4573-4580.  1994.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/ja00090a005\"\n     onclick=\"log_download('mackay-gerhard-beauregard-maplestone-williams-dissectionofthecontributionstowarddimerizationofglycopeptideantibiotics-1994', 'http://doi.org/10.1021/ja00090a005', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-gerhard-beauregard-maplestone-williams-dissectionofthecontributionstowarddimerizationofglycopeptideantibiotics-1994\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>1 download</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-gerhard-beauregard-maplestone-williams-dissectionofthecontributionstowarddimerizationofglycopeptideantibiotics-1994')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Dissection of the Contributions toward Dimerization of Glycopeptide Antibiotics},\n type = {article},\n year = {1994},\n pages = {4573-4580},\n volume = {116},\n id = {5793ec9e-c2d9-3eba-9500-f54fc95f52df},\n created = {2023-01-10T01:47:02.045Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:47:02.045Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {A procedure for the determination of association constants in aqueous solution using hydrogen-deuterium exchange has been developed and used to measure the dimerization constant, Kam, for a number of strongly dimerizing glycopeptide antibiotics. These values provide further insight into the thermodynamic contributions of various structural epitopes to the dimerization of these antibiotics. Consideration of ligand binding affinities together with dimerization potentials provides evidence that dimerization is implicated in the physiological mode of action of these antibiotics. © 1994, American Chemical Society. All rights reserved.},\n bibtype = {article},\n author = {Mackay, J.P. and Gerhard, U. and Beauregard, D.A. and Maplestone, R.A. and Williams, D.H.},\n doi = {10.1021/ja00090a005},\n journal = {Journal of the American Chemical Society},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  A procedure for the determination of association constants in aqueous solution using hydrogen-deuterium exchange has been developed and used to measure the dimerization constant, Kam, for a number of strongly dimerizing glycopeptide antibiotics. These values provide further insight into the thermodynamic contributions of various structural epitopes to the dimerization of these antibiotics. Consideration of ligand binding affinities together with dimerization potentials provides evidence that dimerization is implicated in the physiological mode of action of these antibiotics. © 1994, American Chemical Society. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"mackay-gerhard-beauregard-westwell-searle-williams-glycopeptideantibioticactivityandthepossibleroleofdimerizationamodelforbiologicalsignaling-1994\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Glycopeptide Antibiotic Activity and the Possible Role of Dimerization: A Model for Biological Signaling.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Gerhard,  U.; Beauregard,  D.; Westwell,  M.; Searle,  M.;  and Williams,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of the American Chemical Society</i>, 116(11): 4581-4590.  1994.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/ja00090a006\"\n     onclick=\"log_download('mackay-gerhard-beauregard-westwell-searle-williams-glycopeptideantibioticactivityandthepossibleroleofdimerizationamodelforbiologicalsignaling-1994', 'http://doi.org/10.1021/ja00090a006', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/mackay-gerhard-beauregard-westwell-searle-williams-glycopeptideantibioticactivityandthepossibleroleofdimerizationamodelforbiologicalsignaling-1994\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('mackay-gerhard-beauregard-westwell-searle-williams-glycopeptideantibioticactivityandthepossibleroleofdimerizationamodelforbiologicalsignaling-1994')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Glycopeptide Antibiotic Activity and the Possible Role of Dimerization: A Model for Biological Signaling},\n type = {article},\n year = {1994},\n pages = {4581-4590},\n volume = {116},\n id = {71f945c5-5554-3649-bfe7-e37debf946b9},\n created = {2023-01-10T01:47:02.960Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:47:02.960Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {It is demonstrated that the presence of bacterial cell wall analogues may either enhance or, in the case of ristocetin A, oppose dimerization of glycopeptide antibiotics. These observations may imply that dimerization plays a role in the mode of action of these antibiotics, and a mechanism is proposed to take account of this possibility. The glycopeptide dimers are also found to be formed more exothermically in the presence of cell wall analogues, and the nature of biological signaling events is discussed in this context. It is pointed out that binding enthalpy (rather than simply binding free energy, ΔG) may be an important quantity in signaling events. If this is so, then oligomers may be abundant in signaling processes partly because the extended aggregates they form are able to cooperatively amplify the conformational changes which are incurred on ligand binding, which occur through relatively small changes in free energy but larger opposing changes in enthalpy and entropy. © 1994, American Chemical Society. All rights reserved.},\n bibtype = {article},\n author = {Mackay, J.P. and Gerhard, U. and Beauregard, D.A. and Westwell, M.S. and Searle, M.S. and Williams, D.H.},\n doi = {10.1021/ja00090a006},\n journal = {Journal of the American Chemical Society},\n number = {11}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  It is demonstrated that the presence of bacterial cell wall analogues may either enhance or, in the case of ristocetin A, oppose dimerization of glycopeptide antibiotics. These observations may imply that dimerization plays a role in the mode of action of these antibiotics, and a mechanism is proposed to take account of this possibility. The glycopeptide dimers are also found to be formed more exothermically in the presence of cell wall analogues, and the nature of biological signaling events is discussed in this context. It is pointed out that binding enthalpy (rather than simply binding free energy, ΔG) may be an important quantity in signaling events. If this is so, then oligomers may be abundant in signaling processes partly because the extended aggregates they form are able to cooperatively amplify the conformational changes which are incurred on ligand binding, which occur through relatively small changes in free energy but larger opposing changes in enthalpy and entropy. © 1994, American Chemical Society. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"groves-searle-mackay-williams-thestructureofanasymmetricdimerrelevanttothemodeofactionoftheglycopeptideantibiotics-1994\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The structure of an asymmetric dimer relevant to the mode of action of the glycopeptide antibiotics.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Groves,  P.; Searle,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>;  and Williams,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Structure</i>, 2(8): 747-754.  1994.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1016/S0969-2126(94)00075-1\"\n     onclick=\"log_download('groves-searle-mackay-williams-thestructureofanasymmetricdimerrelevanttothemodeofactionoftheglycopeptideantibiotics-1994', 'http://doi.org/10.1016/S0969-2126(94)00075-1', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/groves-searle-mackay-williams-thestructureofanasymmetricdimerrelevanttothemodeofactionoftheglycopeptideantibiotics-1994\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('groves-searle-mackay-williams-thestructureofanasymmetricdimerrelevanttothemodeofactionoftheglycopeptideantibiotics-1994')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The structure of an asymmetric dimer relevant to the mode of action of the glycopeptide antibiotics},\n type = {article},\n year = {1994},\n pages = {747-754},\n volume = {2},\n id = {19ebea8a-a41d-36ba-9081-a4f636b1f554},\n created = {2023-01-10T01:47:03.895Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:47:03.895Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Background Glycopeptide antibiotics of the vancomycin group are of crucial clinical importance in the treatment of methicillin resistant Staphylococcus aureus (MRSA) - the often lethal &#x27;super-bug &#x27; - characterized by its resistance to a wide range of antibiotics in common use. The antibiotics exert their physiological action by blocking cell wall synthesis through recognition of nascent cell wall mucopeptides terminating in the sequence -D-Ala-D -Ala. Evidence suggests that the antibiotics are able to enhance their biological activity by the formation of homodimers, and this is supported by the observation that dimerization and peptide binding in vitro are cooperative phenomena. The basis of this enhancement is not understood at the molecular level. Results The first detailed structure of a dimeric glycopeptide antibiotic, that of eremomycin, is presented based upon solution NMR data. The overall structure of the dimer complex is asymmetric. The source of this asymmetry - a parallel alignment and mutual interaction of the disaccharides - appears to promote dimerization through specific sugar- sugar recognition. Conclusions A molecular basis for the observed cooperativity of cell wall peptide binding by eremomycin is evident from these studies of the dimer. The carboxylate anion of the cell wall component, which is crucial to binding, forms an amide-mediated ion-pair interaction to the alkylammonium ion of the ring 6 sugar in the other half of the dimer making the structure and positioning of this sugar important in mediating cooperativity. © 1994 Elsevier Science Ltd. All rights reserved.},\n bibtype = {article},\n author = {Groves, P. and Searle, M.S. and Mackay, J.P. and Williams, D.H.},\n doi = {10.1016/S0969-2126(94)00075-1},\n journal = {Structure},\n number = {8}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Background Glycopeptide antibiotics of the vancomycin group are of crucial clinical importance in the treatment of methicillin resistant Staphylococcus aureus (MRSA) - the often lethal 'super-bug ' - characterized by its resistance to a wide range of antibiotics in common use. The antibiotics exert their physiological action by blocking cell wall synthesis through recognition of nascent cell wall mucopeptides terminating in the sequence -D-Ala-D -Ala. Evidence suggests that the antibiotics are able to enhance their biological activity by the formation of homodimers, and this is supported by the observation that dimerization and peptide binding in vitro are cooperative phenomena. The basis of this enhancement is not understood at the molecular level. Results The first detailed structure of a dimeric glycopeptide antibiotic, that of eremomycin, is presented based upon solution NMR data. The overall structure of the dimer complex is asymmetric. The source of this asymmetry - a parallel alignment and mutual interaction of the disaccharides - appears to promote dimerization through specific sugar- sugar recognition. Conclusions A molecular basis for the observed cooperativity of cell wall peptide binding by eremomycin is evident from these studies of the dimer. The carboxylate anion of the cell wall component, which is crucial to binding, forms an amide-mediated ion-pair interaction to the alkylammonium ion of the ring 6 sugar in the other half of the dimer making the structure and positioning of this sugar important in mediating cooperativity. © 1994 Elsevier Science Ltd. All rights reserved.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n  <div class=\"bibbase_group_whole\" id=\"group_1993\">\n    <div class=\"bibbase_group\"\n      onclick=\"toggleGroup('group_1993')\"\n      onkeypress=\"toggleGroup('group_1993')\"\n      tabindex=\"0\">\n      <i class=\"fa fa-angle-down bibbase_group_head_icon\"></i>&nbsp;\n      <span>1993</span>\n      <span class=\"bibbase_group_count\">\n        \n        (3)\n        \n      </span>\n    </div>\n    <div class=\"bibbase_group_body\">\n      \n  \n  <div class=\"bibbase_paper\" id=\"williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  <a href=\"http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172\"\n     onclick=\"return trackOutboundLink('publications', 'click', 'williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993', 'http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172', event);\">\n    Toward an estimation of binding constants in aqueous solution: studies of associations of vancomycin group antibiotics.\n  </a>\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Williams,  D., H.; Searle,  M., S.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J., P.</a>; Gerhard,  U.;  and Maplestone,  R., A.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences</i>, 90(4): 1172-1178. 2 1993.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n  <a href=\"http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172\"\n     onclick=\"log_download('williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993', 'http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172', event.ctrlKey, event)\">\n    <img src=\"//bibbase.org/img/filetypes/link.svg\"\n\t     alt=\"Toward an estimation of binding constants in aqueous solution: studies of associations of vancomycin group antibiotics. [link]\"\n       title=\"Website\"\n\t     class=\"bibbase_icon\"\n         ><span class=\"bibbase_icon_text\">Website</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.90.4.1172\"\n     onclick=\"log_download('williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993', 'http://doi.org/10.1073/pnas.90.4.1172', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Toward an estimation of binding constants in aqueous solution: studies of associations of vancomycin group antibiotics.},\n type = {article},\n year = {1993},\n keywords = {Amide-amide hydrogen bond strengths,Enthalpic barriers,Enthalpy,Entropy compensation,Hydrophobic effect,Rotor restrictions},\n pages = {1172-1178},\n volume = {90},\n websites = {http://www.pnas.org/cgi/doi/10.1073/pnas.90.4.1172},\n month = {2},\n day = {15},\n id = {5ef1cb1b-6f04-35c7-955d-7c5ca2d128fe},\n created = {2020-12-17T05:29:58.125Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2020-12-17T05:29:58.125Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {true},\n hidden = {false},\n citation_key = {Williams1993},\n source_type = {ARTICLE},\n notes = {cited By 160},\n private_publication = {false},\n abstract = {An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.},\n bibtype = {article},\n author = {Williams, Dudley H. and Searle, Mark S. and Mackay, Joel P. and Gerhard, Ute and Maplestone, Rachel A.},\n doi = {10.1073/pnas.90.4.1172},\n journal = {Proceedings of the National Academy of Sciences},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"gerhard-mackay-maplestone-williams-theroleofthesugarandchlorinesubstituentsinthedimerizationofvancomycinantibiotics-1993\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  The Role of the Sugar and Chlorine Substituents in the Dimerization of Vancomycin Antibiotics.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Gerhard,  U.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Maplestone,  R.;  and Williams,  D.\n</span>\n\n  \n\n</span>\n\n  <i>Journal of the American Chemical Society</i>, 115(1): 232-237.  1993.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1021/ja00054a033\"\n     onclick=\"log_download('gerhard-mackay-maplestone-williams-theroleofthesugarandchlorinesubstituentsinthedimerizationofvancomycinantibiotics-1993', 'http://doi.org/10.1021/ja00054a033', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/gerhard-mackay-maplestone-williams-theroleofthesugarandchlorinesubstituentsinthedimerizationofvancomycinantibiotics-1993\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n  &nbsp;\n  <span class=\"bibbase_stats_paper\" style=\"color: #777;\">\n    <span>3 downloads</span>\n  </span>\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('gerhard-mackay-maplestone-williams-theroleofthesugarandchlorinesubstituentsinthedimerizationofvancomycinantibiotics-1993')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {The Role of the Sugar and Chlorine Substituents in the Dimerization of Vancomycin Antibiotics},\n type = {article},\n year = {1993},\n pages = {232-237},\n volume = {115},\n id = {26477609-41c1-3bb1-b211-10fe3f017959},\n created = {2023-01-10T01:47:04.826Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:47:04.826Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {Evidence is presented for the formation of dimers in aqueous solutions of the glycopeptide antibiotics eremomycin, A82846B, vancomycin, and eremomycin-ψ. The dimerization constant Kdim is determined by 1H NMR spectroscopy for the last two compounds and also for the related compound ristocetin-ψ, for which dimerization has previously been reported in mixed solvents. Values of Kdim are obtained for these compounds over a range of temperatures, and thus ΔHdim and ΔSdim are calculated. In addition, a lower limit for Kdim in the case of eremomycin is calculated (105 M−1). This is a remarkably large value, and it may be that dimerization is implicated in antibiotic action. The possibility that natural selection has led to adaptations which promote dimerization (such as the nature and sites of attachment of the sugars and a ring 2 chlorine atom) is discussed. © 1993, American Chemical Society. All rights reserved.},\n bibtype = {article},\n author = {Gerhard, U. and Mackay, J.P. and Maplestone, R.A. and Williams, D.H.},\n doi = {10.1021/ja00054a033},\n journal = {Journal of the American Chemical Society},\n number = {1}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  Evidence is presented for the formation of dimers in aqueous solutions of the glycopeptide antibiotics eremomycin, A82846B, vancomycin, and eremomycin-ψ. The dimerization constant Kdim is determined by 1H NMR spectroscopy for the last two compounds and also for the related compound ristocetin-ψ, for which dimerization has previously been reported in mixed solvents. Values of Kdim are obtained for these compounds over a range of temperatures, and thus ΔHdim and ΔSdim are calculated. In addition, a lower limit for Kdim in the case of eremomycin is calculated (105 M−1). This is a remarkably large value, and it may be that dimerization is implicated in antibiotic action. The possibility that natural selection has led to adaptations which promote dimerization (such as the nature and sites of attachment of the sugars and a ring 2 chlorine atom) is discussed. © 1993, American Chemical Society. All rights reserved.\n</div>\n\n\n</div>\n\n\n  <div class=\"bibbase_paper\" id=\"williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993\">\n  <span class=\"bibbase_paper_titleauthoryear\">\n\n  \n  <span class=\"bibbase_paper_title\">\n  \n  \n  \n  Toward an estimation of binding constants in aqueous solution: Studies of associations of vancomycin group antibiotics.\n  \n  \n  \n</span>\n\n  <span class=\"bibbase_paper_author\">\n  Williams,  D.; Searle,  M.; <a class=\"bibbase author link\" href=\"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386/group/bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2\">Mackay,  J.</a>; Gerhard,  U.;  and Maplestone,  R.\n</span>\n\n  \n\n</span>\n\n  <i>Proceedings of the National Academy of Sciences of the United States of America</i>, 90(4): 1172-1178.  1993.\n  <span class=\"note\"></span>\n\n<span class=\"bibbase_note\"></span>\n\n<br class=\"bibbase_paper_content\"/>\n\n<span class=\"bibbase_paper_content dontprint\">\n\n  \n\n  \n  <a href=\"http://doi.org/10.1073/pnas.90.4.1172\"\n     onclick=\"log_download('williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993', 'http://doi.org/10.1073/pnas.90.4.1172', event.ctrlKey)\"\n     class=\"bibbase doi link\">\n    <span>doi</span></a>\n  &nbsp;\n  \n\n  \n  <a href=\"//bibbase.org/network/publication/williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993\"\n     class=\"bibbase bibbase_page link\">link</a>\n  &nbsp;\n  \n\n  <a href=\"#\"\n     onclick=\"showBib(event); return false;\"\n     class=\"bibbase bibtex link\">bibtex\n    <i class=\"fa fa-caret-down\"></i></a>\n\n  \n  &nbsp;\n  <a class=\"bibbase_abstract_link bibbase link\"\n     href=\"#\"\n     onclick=\"showAbstract(event); return false;\">\n    abstract <i class=\"fa fa-caret-down\"></i></a>\n  \n\n  \n\n  <!-- <a class=\"bibbase read link hidden\" -->\n  <!--    href=\"javascript:toggleRead('williams-searle-mackay-gerhard-maplestone-towardanestimationofbindingconstantsinaqueoussolutionstudiesofassociationsofvancomycingroupantibiotics-1993')\" -->\n  <!--    title=\"Marking this paper as read adds it to your library on BibBase.\" -->\n  <!--    > -->\n  <!--   <i class=\"fa fa-check\"></i>&nbsp; -->\n  <!--   <span>mark as read</span> -->\n  <!-- </a> -->\n\n  &nbsp;\n  \n  \n\n</span>\n\n<div class=\"well well-small bibbase\" data-type=\"bibtex\"\n     style=\"display:none\">\n  <pre style=\"white-space: pre-wrap;\">@article{\n title = {Toward an estimation of binding constants in aqueous solution: Studies of associations of vancomycin group antibiotics},\n type = {article},\n year = {1993},\n pages = {1172-1178},\n volume = {90},\n id = {6e99bb91-e44e-3b3e-a076-022d9dbbba8a},\n created = {2023-01-10T01:47:05.775Z},\n file_attached = {false},\n profile_id = {a5a2ab6f-a8b5-3db6-97bc-618752ee4386},\n group_id = {bc1ab1d4-9e57-37e6-9fb5-435fca0ee9d2},\n last_modified = {2023-01-10T01:47:05.775Z},\n read = {false},\n starred = {false},\n authored = {false},\n confirmed = {false},\n hidden = {false},\n private_publication = {false},\n abstract = {An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.},\n bibtype = {article},\n author = {Williams, D.H. and Searle, M.S. and Mackay, J.P. and Gerhard, U. and Maplestone, R.A.},\n doi = {10.1073/pnas.90.4.1172},\n journal = {Proceedings of the National Academy of Sciences of the United States of America},\n number = {4}\n}</pre>\n</div>\n\n\n<div class=\"well well-small bibbase\" data-type=\"abstract\"\n     style=\"display:none\">\n  An approach toward the estimation of binding constants for organic molecules in aqueous solution is presented, based upon a partitioning of the free energy of binding. Consideration is given to polar and hydrophobic contributions and to the entropic cost of rotor restrictions and bimolecular associations. Several parameters (derived from an analysis of entropy changes upon the melting of crystals and from the binding of cell wall peptide analogues to the antibiotic ristocetin A) which may be useful guides to a crude understanding of binding phenomena are presented: (i) amide-amide hydrogen bond strengths of -(1 to 7) ± 2 kJ·mol-1, (ii) a hydrophobic effect of -0.2 ± 0.05 kJ·mol-1·Å-2 of hydrocarbon removed from exposure to water in the binding process, and (iii) free energy costs for rotor restrictions of 3.5-5.0 kJ·mol-1. The validity of the parameters for hydrogen bond strengths is dependent on the validity of the other two parameters. The phenomenon of entropy/enthalpy compensation is considered, with the conclusion that enthalpic barriers to dissociations will result in larger losses in translational and rotational entropy in the association step. The dimerization of some vancomycin group antibiotics is strongly exothermic (-36 to -51 kJ·mol-1) and is promoted by a factor of 50-100 by a disaccharide attached to ring 4 (in vancomycin and eremomycin) and by a factor of ca. 1000 by an amino-sugar attached to the benzylic position of ring 6 in eremomycin. The dimerization process (which, as required for an exothermic association, appears to be costly in entropy) may be relevant to the mode of action of the antibiotics.\n</div>\n\n\n</div>\n\n\n\n\n\n    </div>\n  </div>\n\n\n\n\n  </div>\n\n\n  <!-- Modal -->\n\n  <!-- <iframe id=\"bibbase_network_frame\" -->\n  <!--         src=\"//bibbase.org/network/control\" -->\n  <!--         style=\"display: none;\"> -->\n  <!-- </iframe> -->\n\n</div>\n"}; document.write(bibbase_data.data);