TY - JOUR
T1 - Molecular determinants of the functional interaction between syntaxin and N-type Ca2+ channel gating
AU - Bezprozvanny, Ilya
AU - Zhong, Pingyu
AU - Scheller, Richard H.
AU - Tsien, Richard W.
PY - 2000/12/5
Y1 - 2000/12/5
N2 - Syntaxin is a key presynaptic protein that binds to N- and P/Q-type Ca2+ channels in biochemical studies and affects gating of these Ca2+ channels in expression systems and in synaptosomes. The present study was aimed at understanding the molecular basis of syntaxin modulation of N-type channel gating. Mutagenesis of either syntaxin 1A or the pore-forming α(1B) subunit of N-type Ca2+ channels was combined with functional assays of N-type channel gating in a Xenopus oocyte coexpression system and in biochemical binding experiments in vitro. Our analysis showed that the transmembrane region of syntaxin and a short region within the H3 helical cytoplasmic domain of syntaxin, containing residues Ala-240 and Val-244, appeared critical for the channel modulation but not for biochemical association with the 'synprint site' in the II/III loop of α(1B). These results suggest that syntaxin and the α(1B) subunit engage in two kinds of interactions: an anchoring interaction via the II/III loop synprint site and a modulatory interaction via another site located elsewhere in the channel sequence. The segment of syntaxin H3 found to be involved in the modulatory interaction would lie hidden within the four-helix structure of the SNARE complex, supporting the hypothesis that syntaxin's ability to regulate N-type Ca2+ channels would be enabled after SNARE complex disassembly after synaptic vesicle exocytosis.
AB - Syntaxin is a key presynaptic protein that binds to N- and P/Q-type Ca2+ channels in biochemical studies and affects gating of these Ca2+ channels in expression systems and in synaptosomes. The present study was aimed at understanding the molecular basis of syntaxin modulation of N-type channel gating. Mutagenesis of either syntaxin 1A or the pore-forming α(1B) subunit of N-type Ca2+ channels was combined with functional assays of N-type channel gating in a Xenopus oocyte coexpression system and in biochemical binding experiments in vitro. Our analysis showed that the transmembrane region of syntaxin and a short region within the H3 helical cytoplasmic domain of syntaxin, containing residues Ala-240 and Val-244, appeared critical for the channel modulation but not for biochemical association with the 'synprint site' in the II/III loop of α(1B). These results suggest that syntaxin and the α(1B) subunit engage in two kinds of interactions: an anchoring interaction via the II/III loop synprint site and a modulatory interaction via another site located elsewhere in the channel sequence. The segment of syntaxin H3 found to be involved in the modulatory interaction would lie hidden within the four-helix structure of the SNARE complex, supporting the hypothesis that syntaxin's ability to regulate N-type Ca2+ channels would be enabled after SNARE complex disassembly after synaptic vesicle exocytosis.
UR - http://www.scopus.com/inward/record.url?scp=0034610298&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034610298&partnerID=8YFLogxK
U2 - 10.1073/pnas.220389697
DO - 10.1073/pnas.220389697
M3 - Article
C2 - 11087812
AN - SCOPUS:0034610298
SN - 0027-8424
VL - 97
SP - 13943
EP - 13948
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 25
ER -