Complexin arrests a pool of docked vesicles for fast Ca2+ -dependent release

Regulated exocytosis requires that the assembly of the basic membrane fusion machinery is temporarily arrested. Synchronized membrane fusion is then caused by a specific trigger—a local rise of the Ca2+ concentration. Using reconstituted giant unilamellar vesicles (GUVs), we have analysed the role o...

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Bibliographic Details
Main Authors: Malsam, Jörg (Author) , Parisotto, Daniel (Author) , Malsam, Andrea (Author) , Krause, Jean Michel (Author) , Söllner, Thomas (Author)
Format: Article (Journal)
Language:English
Published: 15. June 2012
In: The EMBO journal
Year: 2012, Volume: 31, Issue: 15, Pages: 3270-3281
ISSN:1460-2075
DOI:10.1038/emboj.2012.164
Online Access:Verlag, Volltext: http://dx.doi.org/10.1038/emboj.2012.164
Verlag, Volltext: http://emboj.embopress.org/content/31/15/3270
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Author Notes:Jörg Malsam, Daniel Parisotto, Tanmay A. M. Bharat, Andrea Scheutzow, Jean Michel Krause, John A. G. Briggs and Thomas H. Söllner
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Summary:Regulated exocytosis requires that the assembly of the basic membrane fusion machinery is temporarily arrested. Synchronized membrane fusion is then caused by a specific trigger—a local rise of the Ca2+ concentration. Using reconstituted giant unilamellar vesicles (GUVs), we have analysed the role of complexin and membrane‐anchored synaptotagmin 1 in arresting and synchronizing fusion by lipid‐mixing and cryo‐electron microscopy. We find that they mediate the formation and consumption of docked small unilamellar vesicles (SUVs) via the following sequence of events: Synaptotagmin 1 mediates v‐SNARE‐SUV docking to t‐SNARE‐GUVs in a Ca2+‐independent manner. Complexin blocks vesicle consumption, causing accumulation of docked vesicles. Together with synaptotagmin 1, complexin synchronizes and stimulates rapid fusion of accumulated docked vesicles in response to physiological Ca2+ concentrations. Thus, the reconstituted assay resolves both the stimulatory and inhibitory function of complexin and mimics key aspects of synaptic vesicle fusion.
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Physical Description:Online Resource
ISSN:1460-2075
DOI:10.1038/emboj.2012.164