@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}
}

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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},"bibtype":"article","creationDate":"2020-07-10T05:31:56.153Z","downloads":0,"keywords":[],"search_terms":["terminal","region","stalk","domain","ubiquitous","human","kinesin","heavy","chain","contains","binding","site","kinesin","light","chain","diefenbach","mackay","armati","cunningham"],"title":"The C-terminal region of the stalk domain of ubiquitous human kinesin heavy chain contains the binding site for kinesin light chain","year":1998,"biburl":"https://bibbase.org/service/mendeley/a5a2ab6f-a8b5-3db6-97bc-618752ee4386","dataSources":["QNo3CKhtog6c3sh7Y","ya2CyA73rpZseyrZ8","4BKYoCQJ3ndMcbbtH","2252seNhipfTmjEBQ","ibLPPAndxdcRpxcwH"]}