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

Malsam, J.; Parisotto, D.; Bharat, T. A. M.; Scheutzow, A.; Krause, J. M.; Briggs, John A. G.; Sollner, T. H.

doi:10.1038/emboj.2012.164

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

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Malsam, J., Parisotto, D., Bharat, T. A. M., Scheutzow, A., Krause, J. M., Briggs, J. A. G., et al. (2012). Complexin arrests a pool of docked vesicles for fast Ca2+-dependent release. Embo Journal, 31(15), 3270-3281. doi:10.1038/emboj.2012.164.

Cite as: https://hdl.handle.net/21.11116/0000-0009-70FA-F

Abstract

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. The EMBO Journal (2012) 31, 3270-3281. doi:10.1038/emboj.2012.164; Published online 15 June 2012