Molecular determinants of the functional interaction between syntaxin and N-type Ca2+ channel gating

Ilya Bezprozvanny, Pingyu Zhong, Richard H. Scheller, Richard W. Tsien

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)13943-13948
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume97
Issue number25
DOIs
StatePublished - Dec 5 2000

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Qa-SNARE Proteins
Syntaxin 1
SNARE Proteins
Aptitude
Synaptosomes
Synaptic Vesicles
Exocytosis
Xenopus
Mutagenesis
Oocytes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Molecular determinants of the functional interaction between syntaxin and N-type Ca2+ channel gating. / Bezprozvanny, Ilya; Zhong, Pingyu; Scheller, Richard H.; Tsien, Richard W.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, No. 25, 05.12.2000, p. 13943-13948.

Research output: Contribution to journalArticle

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