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PMID: 20406821 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A coiled coil trigger site is essential for rapid binding of synaptobrevin to the SNARE acceptor complex.

The Journal of biological chemistry ·Vol. 285 ·No. 28 ·2010-07-09 ·Pages 21549-59

Wiederhold K, Kloepper TH, Walter AM, Stein A, Kienle N, Sørensen JB, Fasshauer D

Abstract

Exocytosis from synaptic vesicles is driven by stepwise formation of a tight alpha-helical complex between the fusing membranes. The complex is composed of the three SNAREs: synaptobrevin 2, SNAP-25, and syntaxin 1a. An important step in complex formation is fast binding of vesicular synaptobrevin to the preformed syntaxin 1.SNAP-25 dimer. Exactly how this step relates to neurotransmitter release is not well understood. Here, we combined different approaches to gain insights into this reaction. Using computational methods, we identified a stretch in synaptobrevin 2 that may function as a coiled coil "trigger site." This site is also present in many synaptobrevin homologs functioning in other trafficking steps. Point mutations in this stretch inhibited binding to the syntaxin 1.SNAP-25 dimer and slowed fusion of liposomes. Moreover, the point mutations severely inhibited secretion from chromaffin cells. Altogether, this demonstrates that the trigger site in synaptobrevin is crucial for productive SNARE zippering.

MeSH Terms
Amino Acid Motifs Animals Binding Sites Calcium/chemistry Calorimetry/methods Chromaffin Cells/metabolism Dimerization Electrophysiology/methods Liposomes/chemistry Mice Neurotransmitter Agents/metabolism Point Mutation Protein Structure, Tertiary R-SNARE Proteins/chemistry Rats SNARE Proteins/chemistry
Chemicals
Liposomes Neurotransmitter Agents R-SNARE Proteins SNARE Proteins Calcium
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Wiederhold Katrin
Research Group Structural Biochemistry, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
Kloepper Tobias H
Walter Alexander M
Stein Alexander
Kienle Nickias
Sørensen Jakob B
Fasshauer Dirk
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M110.105148
Published
2010-07-09
Epub
2010-00-20
Pages
21549-59
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2898431
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M110.105148 PMC Full Text View journal details
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