Structure and lipid-mediated remodelling mechanism of the Na+/H+ exchanger NHA2. Matsuoka, R., Fudim, R., Jung, S., Zhang, C., Bazzone, A., Chatzikyriakidou, Y., Nomura, N., Iwata, S., Orellana, L., Beckstein, O., & Drew, D. Technical Report July, 2021. Company: Cold Spring Harbor Laboratory Distributor: Cold Spring Harbor Laboratory Label: Cold Spring Harbor Laboratory Section: New Results Type: article![Structure and lipid-mediated remodelling mechanism of the Na+/H+ exchanger NHA2 [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper doi abstract bibtex Na+/H+ exchangers catalyse an ion-exchange activity that is carried out in most, if not all cells. SLC9B2, also known as NHA2, correlates with the long-sought after sodium/lithium (Na+/Li+) exchanger linked to the pathogenesis of diabetes mellitus and essential hypertension in humans. Despite its functional importance, structural information and the molecular basis of its ion-exchange mechanism have been lacking. Here, we report the cryo EM structures of bison NHA2 in detergent and in nanodiscs at 3.0 and 3.5 Å resolution, respectively. NHA2 shares closest structural similarity to the bacterial electrogenic Na+/H+ antiporter NapA, rather than other mammalian SLC9A members. Nevertheless, SSM-based electrophysiology results with NHA2 show the catalysis of electroneutral rather than electrogenic ion exchange, and the ion-binding site is quite distinctive, with a tryptophan-arginine- glutamate triad separated from the well-established ion-binding aspartates. These triad residues fine-tune ion binding specificity, as demonstrated by a salt-bridge swap mutant that converts NHA2 into a Li+-specific transporter. Strikingly, an additional N-terminal helix in NHA2 establishes a unique homodimer with a large ∼ 25 Å intracellular gap between protomers. In the presence of phosphatidylinositol lipids, the N-terminal helix rearranges and closes this gap. We confirm that dimerization of NHA2 is required for activity in vivo, and propose that the N- terminal helix has evolved as a lipid-mediated remodelling switch for regulation of transport activity.
@techreport{matsuoka_structure_2021,
title = {Structure and lipid-mediated remodelling mechanism of the {Na}+/{H}+ exchanger {NHA2}},
copyright = {© 2021, Posted by Cold Spring Harbor Laboratory. The copyright holder for this pre-print is the author. All rights reserved. The material may not be redistributed, re-used or adapted without the author's permission.},
url = {https://www.biorxiv.org/content/10.1101/2021.07.22.453398v1},
abstract = {Na+/H+ exchangers catalyse an ion-exchange activity that is carried out in most, if not all cells. SLC9B2, also known as NHA2, correlates with the long-sought after sodium/lithium (Na+/Li+) exchanger linked to the pathogenesis of diabetes mellitus and essential hypertension in humans. Despite its functional importance, structural information and the molecular basis of its ion-exchange mechanism have been lacking. Here, we report the cryo EM structures of bison NHA2 in detergent and in nanodiscs at 3.0 and 3.5 Å resolution, respectively. NHA2 shares closest structural similarity to the bacterial electrogenic Na+/H+ antiporter NapA, rather than other mammalian SLC9A members. Nevertheless, SSM-based electrophysiology results with NHA2 show the catalysis of electroneutral rather than electrogenic ion exchange, and the ion-binding site is quite distinctive, with a tryptophan-arginine- glutamate triad separated from the well-established ion-binding aspartates. These triad residues fine-tune ion binding specificity, as demonstrated by a salt-bridge swap mutant that converts NHA2 into a Li+-specific transporter. Strikingly, an additional N-terminal helix in NHA2 establishes a unique homodimer with a large ∼ 25 Å intracellular gap between protomers. In the presence of phosphatidylinositol lipids, the N-terminal helix rearranges and closes this gap. We confirm that dimerization of NHA2 is required for activity in vivo, and propose that the N- terminal helix has evolved as a lipid-mediated remodelling switch for regulation of transport activity.},
language = {en},
urldate = {2021-08-31},
author = {Matsuoka, Rei and Fudim, Roman and Jung, Sukkyeong and Zhang, Chenou and Bazzone, Andre and Chatzikyriakidou, Yurie and Nomura, Norimichi and Iwata, So and Orellana, Laura and Beckstein, Oliver and Drew, David},
month = jul,
year = {2021},
doi = {10.1101/2021.07.22.453398},
note = {Company: Cold Spring Harbor Laboratory
Distributor: Cold Spring Harbor Laboratory
Label: Cold Spring Harbor Laboratory
Section: New Results
Type: article},
pages = {2021.07.22.453398},
}
Downloads: 0
{"_id":"Gnh7Au2hZXoreJyC3","bibbaseid":"matsuoka-fudim-jung-zhang-bazzone-chatzikyriakidou-nomura-iwata-etal-structureandlipidmediatedremodellingmechanismofthenahexchangernha2-2021","author_short":["Matsuoka, R.","Fudim, R.","Jung, S.","Zhang, C.","Bazzone, A.","Chatzikyriakidou, Y.","Nomura, N.","Iwata, S.","Orellana, L.","Beckstein, O.","Drew, D."],"bibdata":{"bibtype":"techreport","type":"techreport","title":"Structure and lipid-mediated remodelling mechanism of the Na+/H+ exchanger NHA2","copyright":"© 2021, Posted by Cold Spring Harbor Laboratory. The copyright holder for this pre-print is the author. All rights reserved. The material may not be redistributed, re-used or adapted without the author's permission.","url":"https://www.biorxiv.org/content/10.1101/2021.07.22.453398v1","abstract":"Na+/H+ exchangers catalyse an ion-exchange activity that is carried out in most, if not all cells. SLC9B2, also known as NHA2, correlates with the long-sought after sodium/lithium (Na+/Li+) exchanger linked to the pathogenesis of diabetes mellitus and essential hypertension in humans. Despite its functional importance, structural information and the molecular basis of its ion-exchange mechanism have been lacking. Here, we report the cryo EM structures of bison NHA2 in detergent and in nanodiscs at 3.0 and 3.5 Å resolution, respectively. NHA2 shares closest structural similarity to the bacterial electrogenic Na+/H+ antiporter NapA, rather than other mammalian SLC9A members. Nevertheless, SSM-based electrophysiology results with NHA2 show the catalysis of electroneutral rather than electrogenic ion exchange, and the ion-binding site is quite distinctive, with a tryptophan-arginine- glutamate triad separated from the well-established ion-binding aspartates. These triad residues fine-tune ion binding specificity, as demonstrated by a salt-bridge swap mutant that converts NHA2 into a Li+-specific transporter. Strikingly, an additional N-terminal helix in NHA2 establishes a unique homodimer with a large ∼ 25 Å intracellular gap between protomers. In the presence of phosphatidylinositol lipids, the N-terminal helix rearranges and closes this gap. We confirm that dimerization of NHA2 is required for activity in vivo, and propose that the N- terminal helix has evolved as a lipid-mediated remodelling switch for regulation of transport activity.","language":"en","urldate":"2021-08-31","author":[{"propositions":[],"lastnames":["Matsuoka"],"firstnames":["Rei"],"suffixes":[]},{"propositions":[],"lastnames":["Fudim"],"firstnames":["Roman"],"suffixes":[]},{"propositions":[],"lastnames":["Jung"],"firstnames":["Sukkyeong"],"suffixes":[]},{"propositions":[],"lastnames":["Zhang"],"firstnames":["Chenou"],"suffixes":[]},{"propositions":[],"lastnames":["Bazzone"],"firstnames":["Andre"],"suffixes":[]},{"propositions":[],"lastnames":["Chatzikyriakidou"],"firstnames":["Yurie"],"suffixes":[]},{"propositions":[],"lastnames":["Nomura"],"firstnames":["Norimichi"],"suffixes":[]},{"propositions":[],"lastnames":["Iwata"],"firstnames":["So"],"suffixes":[]},{"propositions":[],"lastnames":["Orellana"],"firstnames":["Laura"],"suffixes":[]},{"propositions":[],"lastnames":["Beckstein"],"firstnames":["Oliver"],"suffixes":[]},{"propositions":[],"lastnames":["Drew"],"firstnames":["David"],"suffixes":[]}],"month":"July","year":"2021","doi":"10.1101/2021.07.22.453398","note":"Company: Cold Spring Harbor Laboratory Distributor: Cold Spring Harbor Laboratory Label: Cold Spring Harbor Laboratory Section: New Results Type: article","pages":"2021.07.22.453398","bibtex":"@techreport{matsuoka_structure_2021,\n\ttitle = {Structure and lipid-mediated remodelling mechanism of the {Na}+/{H}+ exchanger {NHA2}},\n\tcopyright = {© 2021, Posted by Cold Spring Harbor Laboratory. The copyright holder for this pre-print is the author. All rights reserved. The material may not be redistributed, re-used or adapted without the author's permission.},\n\turl = {https://www.biorxiv.org/content/10.1101/2021.07.22.453398v1},\n\tabstract = {Na+/H+ exchangers catalyse an ion-exchange activity that is carried out in most, if not all cells. SLC9B2, also known as NHA2, correlates with the long-sought after sodium/lithium (Na+/Li+) exchanger linked to the pathogenesis of diabetes mellitus and essential hypertension in humans. Despite its functional importance, structural information and the molecular basis of its ion-exchange mechanism have been lacking. Here, we report the cryo EM structures of bison NHA2 in detergent and in nanodiscs at 3.0 and 3.5 Å resolution, respectively. NHA2 shares closest structural similarity to the bacterial electrogenic Na+/H+ antiporter NapA, rather than other mammalian SLC9A members. Nevertheless, SSM-based electrophysiology results with NHA2 show the catalysis of electroneutral rather than electrogenic ion exchange, and the ion-binding site is quite distinctive, with a tryptophan-arginine- glutamate triad separated from the well-established ion-binding aspartates. These triad residues fine-tune ion binding specificity, as demonstrated by a salt-bridge swap mutant that converts NHA2 into a Li+-specific transporter. Strikingly, an additional N-terminal helix in NHA2 establishes a unique homodimer with a large ∼ 25 Å intracellular gap between protomers. In the presence of phosphatidylinositol lipids, the N-terminal helix rearranges and closes this gap. We confirm that dimerization of NHA2 is required for activity in vivo, and propose that the N- terminal helix has evolved as a lipid-mediated remodelling switch for regulation of transport activity.},\n\tlanguage = {en},\n\turldate = {2021-08-31},\n\tauthor = {Matsuoka, Rei and Fudim, Roman and Jung, Sukkyeong and Zhang, Chenou and Bazzone, Andre and Chatzikyriakidou, Yurie and Nomura, Norimichi and Iwata, So and Orellana, Laura and Beckstein, Oliver and Drew, David},\n\tmonth = jul,\n\tyear = {2021},\n\tdoi = {10.1101/2021.07.22.453398},\n\tnote = {Company: Cold Spring Harbor Laboratory\nDistributor: Cold Spring Harbor Laboratory\nLabel: Cold Spring Harbor Laboratory\nSection: New Results\nType: article},\n\tpages = {2021.07.22.453398},\n}\n\n","author_short":["Matsuoka, R.","Fudim, R.","Jung, S.","Zhang, C.","Bazzone, A.","Chatzikyriakidou, Y.","Nomura, N.","Iwata, S.","Orellana, L.","Beckstein, O.","Drew, D."],"key":"matsuoka_structure_2021","id":"matsuoka_structure_2021","bibbaseid":"matsuoka-fudim-jung-zhang-bazzone-chatzikyriakidou-nomura-iwata-etal-structureandlipidmediatedremodellingmechanismofthenahexchangernha2-2021","role":"author","urls":{"Paper":"https://www.biorxiv.org/content/10.1101/2021.07.22.453398v1"},"metadata":{"authorlinks":{}}},"bibtype":"techreport","biburl":"https://api.zotero.org/users/1446965/collections/GJIR5FQV/items?key=DK7eBbaofVxXe4ShaO2nLItp&format=bibtex&limit=100","dataSources":["aWZX3bdqYnwJ4TFqr","PGB7KMr8nSSyHsKme","4vn2CWL2AgyPs2kny","Y8jAQ6b2eCo2cdomS","nsS5dn53AvB6eMZRk"],"keywords":[],"search_terms":["structure","lipid","mediated","remodelling","mechanism","exchanger","nha2","matsuoka","fudim","jung","zhang","bazzone","chatzikyriakidou","nomura","iwata","orellana","beckstein","drew"],"title":"Structure and lipid-mediated remodelling mechanism of the Na+/H+ exchanger NHA2","year":2021}