Synaptobrevin N-terminally bound to syntaxin–SNAP-25 defines the primed vesicle state in regulated exocytosis
10.1083/jcb.200907018
Crossref journal-article
Rockefeller University Press
Journal of Cell Biology (291)
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 Ca2+. 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 Ca2+-triggered C-terminal assembly and membrane fusion.

Authors 5
  1. Alexander M. Walter (first)
  2. Katrin Wiederhold (additional)
  3. Dieter Bruns (additional)
  4. Dirk Fasshauer (additional)
  5. Jakob B. Sørensen (additional)
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Dates
Type When
Created 15 years, 6 months ago (Feb. 8, 2010, 1:03 p.m.)
Deposited 2 years, 1 month ago (July 29, 2023, 12:57 a.m.)
Indexed 3 months ago (May 28, 2025, 2:02 p.m.)
Issued 15 years, 6 months ago (Feb. 8, 2010)
Published 15 years, 6 months ago (Feb. 8, 2010)
Published Online 15 years, 6 months ago (Feb. 8, 2010)
Published Print 15 years, 6 months ago (Feb. 8, 2010)
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@article{Walter_2010, title={Synaptobrevin N-terminally bound to syntaxin–SNAP-25 defines the primed vesicle state in regulated exocytosis}, volume={188}, ISSN={0021-9525}, url={http://dx.doi.org/10.1083/jcb.200907018}, DOI={10.1083/jcb.200907018}, number={3}, journal={Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Walter, Alexander M. and Wiederhold, Katrin and Bruns, Dieter and Fasshauer, Dirk and Sørensen, Jakob B.}, year={2010}, month=feb, pages={401–413} }