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Fast Vesicle Fusion in Living Cells Requires at Least Three SNARE Complexes

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Science  16 Sep 2010:
1193134
DOI: 10.1126/science.1193134

Abstract

Exocytosis requires formation of SNARE complexes between vesicle- and target-membranes. Recent assessments in reduced model systems have produced divergent estimates of the number of SNARE complexes needed for fusion. Here, we used a titration approach to answer this question in intact, cultured chromaffin cells. Simultaneous expression of wild-type SNAP-25 and a mutant unable to support exocytosis progressively altered fusion kinetics and fusion pore opening, indicating that both proteins assemble into heteromeric fusion complexes. Expressing different wild-type:mutant ratios revealed a third power relationship for fast (synchronous) fusion and a near-linear relationship for overall release. Thus, fast fusion typically observed in synapses and neurosecretory cells requires at least three functional SNARE complexes, while slower release might occur with fewer. Heterogeneity in SNARE-complex number may explain heterogeneity in vesicular release probability.