Synaptobrevin N-terminally bound to syntaxin-SNAP-25 defines the primed vesicle state in regulated exocytosis.

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Version: Final published version
Serval ID
serval:BIB_18235A8E35A3
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Synaptobrevin N-terminally bound to syntaxin-SNAP-25 defines the primed vesicle state in regulated exocytosis.
Journal
Journal of Cell Biology
Author(s)
Walter A.M., Wiederhold K., Bruns D., Fasshauer D., Sørensen J.B.
ISSN
1540-8140 (Electronic)
ISSN-L
0021-9525
Publication state
Published
Issued date
2010
Volume
188
Number
3
Pages
401-413
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
Rapid neurotransmitter release depends on the ability to arrest the SNAP receptor (SNARE)-dependent exocytosis pathway at an intermediate "cocked" state, from which fusion can be triggered by Ca(2+). It is not clear whether this state includes assembly of synaptobrevin (the vesicle membrane SNARE) to the syntaxin-SNAP-25 (target membrane SNAREs) acceptor complex or whether the reaction is arrested upstream of that step. In this study, by a combination of in vitro biophysical measurements and time-resolved exocytosis measurements in adrenal chromaffin cells, we find that mutations of the N-terminal interaction layers of the SNARE bundle inhibit assembly in vitro and vesicle priming in vivo without detectable changes in triggering speed or fusion pore properties. In contrast, mutations in the last C-terminal layer decrease triggering speed and fusion pore duration. Between the two domains, we identify a region exquisitely sensitive to mutation, possibly constituting a switch. Our data are consistent with a model in which the N terminus of the SNARE complex assembles during vesicle priming, followed by Ca(2+)-triggered C-terminal assembly and membrane fusion.
Keywords
Animals, Calcium/metabolism, Exocytosis/physiology, Mice, Mice, Knockout, Models, Biological, Qa-SNARE Proteins/genetics, Qa-SNARE Proteins/metabolism, R-SNARE Proteins/genetics, R-SNARE Proteins/metabolism, Rats, SNARE Proteins/genetics, SNARE Proteins/metabolism, Secretory Vesicles/genetics, Secretory Vesicles/metabolism, Synaptosomal-Associated Protein 25/genetics, Synaptosomal-Associated Protein 25/metabolism
Pubmed
Web of science
Open Access
Yes
Create date
23/03/2011 10:28
Last modification date
20/08/2019 12:48
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