Mechanism of SNARE protein binding and regulation of Ca v2 channels by phosphorylation of the synaptic protein interaction site

Charles T. Yokoyama, Scott J. Myers, Jian Fu, Susan M. Mockus, Todd Scheuer, William A. Catterall

Research output: Contribution to journalArticlepeer-review

75 Scopus citations

Abstract

Ca v2.1 and Ca v2.2 channels conduct P/Q-type and N-type Ca 2+ currents that initiate neurotransmission and bind SNARE proteins through a synaptic protein interaction (synprint) site. PKC and CaMKII phosphorylate the synprint site and inhibit SNARE protein binding in vitro. Here we identify two separate microdomains that each bind syntaxin 1A and SNAP-25 in vitro and are regulated by PKC phosphorylation at serines 774 and 898 and CaMKII phosphorylation at serines 784 and 896. Activation of PKC resulted in its recruitment to and phosphorylation of Ca V2.2 channels, but PKC phosphorylation did not dissociate Ca V2.2 channel/syntaxin 1A complexes. Chimeric Ca V2.1a channels containing the synprint site of Ca v2.2 gain modulation by syntaxin 1A, which is blocked by PKC phosphorylation at the sites identified above. Our results support a bipartite model for the synprint site in which each SNARE-binding microdomain is controlled by a separate PKC and CaMKII phosphorylation site that regulates channel modulation by SNARE proteins.

Original languageEnglish
Pages (from-to)1-17
Number of pages17
JournalMolecular and Cellular Neuroscience
Volume28
Issue number1
DOIs
StatePublished - Jan 2005

Bibliographical note

Funding Information:
We thank Dr. T.P. Snutch (University of British Columbia) for Ca V 2.1 and Ca v 2.2 cDNAs, Dr. M. Wilson (University of New Mexico) for SNAP-25B cDNA, and Dr. Masami Takahashi (Kitasato University) for syntaxin 1A cDNA and Dr. Thomas Soderling (Vollum Institute) for CaMKII. This research was supported by an NRSA from NIH Training Grant T32 NS007332 to S.J.M., NRSA 1F32NS11030 to S.M.M., and NIH Research Grant NS22625 to W.A.C.

Funding

We thank Dr. T.P. Snutch (University of British Columbia) for Ca V 2.1 and Ca v 2.2 cDNAs, Dr. M. Wilson (University of New Mexico) for SNAP-25B cDNA, and Dr. Masami Takahashi (Kitasato University) for syntaxin 1A cDNA and Dr. Thomas Soderling (Vollum Institute) for CaMKII. This research was supported by an NRSA from NIH Training Grant T32 NS007332 to S.J.M., NRSA 1F32NS11030 to S.M.M., and NIH Research Grant NS22625 to W.A.C.

FundersFunder number
National Institutes of Health (NIH)T32 NS007332, 1F32NS11030
National Institute of Neurological Disorders and StrokeR01NS022625
Israel National Road Safety Authority

    ASJC Scopus subject areas

    • Molecular Biology
    • Cellular and Molecular Neuroscience
    • Cell Biology

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