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Science 304 (5669): 435-438

Copyright © 2004 by the American Association for the Advancement of Science

Reconstitution of Ca2+-Regulated Membrane Fusion by Synaptotagmin and SNAREs

Ward C. Tucker,1 Thomas Weber,2 Edwin R. Chapman1*

Abstract: We investigated the effect of synaptotagmin I on membrane fusion mediated by neuronal SNARE proteins, SNAP-25, syntaxin, and synaptobrevin, which were reconstituted into vesicles. In the presence of Ca2+, the cytoplasmic domain of synaptotagmin I (syt) strongly stimulated membrane fusion when synaptobrevin densities were similar to those found in native synaptic vesicles. The Ca2+ dependence of syt-stimulated fusion was modulated by changes in lipid composition of the vesicles and by a truncation that mimics cleavage of SNAP-25 by botulinum neurotoxin A. Stimulation of fusion was abolished by disrupting the Ca2+-binding activity, or by severing the tandem C2 domains, of syt. Thus, syt and SNAREs are likely to represent the minimal protein complement for Ca2+-triggered exocytosis.

1 Department of Physiology, University of Wisconsin, Madison, WI 53706, USA.
2 Carl C. Icahn Center for Gene Therapy and Molecular Medicine and the Department of Molecular, Cell and Developmental Biology, Mount Sinai School of Medicine, New York, NY 10029, USA.

* To whom correspondence should be addressed. E-mail: chapman{at}physiology.wisc.edu

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