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PMID: 20826818 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Novel interactions of CAPS (Ca2+-dependent activator protein for secretion) with the three neuronal SNARE proteins required for vesicle fusion.

The Journal of biological chemistry ·Vol. 285 ·No. 46 ·2010-11-12 ·Pages 35320-9

Daily NJ, Boswell KL, James DJ, Martin TF

Abstract

CAPS (aka CADPS) is required for optimal vesicle exocytosis in neurons and endocrine cells where it functions to prime the exocytic machinery for Ca(2+)-triggered fusion. Fusion is mediated by trans complexes of the SNARE proteins VAMP-2, syntaxin-1, and SNAP-25 that bridge vesicle and plasma membrane. CAPS promotes SNARE complex formation on liposomes, but the SNARE binding properties of CAPS are unknown. The current work revealed that CAPS exhibits high affinity binding to syntaxin-1 and SNAP-25 and moderate affinity binding to VAMP-2. CAPS binding is specific for a subset of exocytic SNARE protein isoforms and requires membrane integration of the SNARE proteins. SNARE protein binding by CAPS is novel and mediated by interactions with the SNARE motifs in the three proteins. The C-terminal site for CAPS binding on syntaxin-1 does not overlap the Munc18-1 binding site and both proteins can co-reside on membrane-integrated syntaxin-1. As expected for a C-terminal binding site on syntaxin-1, CAPS stimulates SNARE-dependent liposome fusion with N-terminal truncated syntaxin-1 but exhibits impaired activity with C-terminal syntaxin-1 mutants. Overall the results suggest that SNARE complex formation promoted by CAPS may be mediated by direct interactions of CAPS with each of the three SNARE proteins required for vesicle exocytosis.

MeSH Terms
Animals Binding, Competitive Calcium-Binding Proteins/chemistry,genetics,metabolism Cell Line HEK293 Cells Humans Immunoblotting Kinetics Liposomes/chemistry,metabolism Membrane Fusion Mice Neurons/metabolism Phosphatidylcholines/chemistry Phosphatidylserines/chemistry Protein Binding Protein Multimerization Proteolipids/chemistry,metabolism Rats Recombinant Proteins/chemistry,metabolism SNARE Proteins/chemistry,genetics,metabolism Spodoptera Synaptosomal-Associated Protein 25/chemistry,genetics,metabolism Syntenins/chemistry,genetics,metabolism Vesicle-Associated Membrane Protein 2/chemistry,genetics,metabolism
Chemicals
Cadps protein, rat Calcium-Binding Proteins Liposomes Phosphatidylcholines Phosphatidylserines Proteolipids Recombinant Proteins SNARE Proteins Synaptosomal-Associated Protein 25 Syntenins Vesicle-Associated Membrane Protein 2 proteoliposomes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Daily Neil J
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Boswell Kristin L
James Declan J
Martin Thomas F J
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M110.145169
Published
2010-11-12
Epub
2010-00-08
Pages
35320-9
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2975156
Subset
IM
Grants
NIDDK NIH HHS · R01 DK040428 · United States
NCRR NIH HHS · S10 RR013790 · United States
External Links
PubMed source DOI: 10.1074/jbc.M110.145169 PMC Full Text View journal details
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