Single SNARE-mediated vesicle fusion observed in vitro by polarized TIRFM.

ABSTRACT: Single-vesicle fusion assays in vitro are useful tools for examining mechanisms of membrane fusion at the molecular level mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs). This approach allows the experimentalist to define the lipid and protein composition of the two fusing membranes and perform experiments under highly controlled conditions. In previous experiments, in which we reconstituted a SNARE acceptor complex into supported membranes and observed the docking and fusion of fluorescently labeled synaptobrevin proteoliposomes by total internal reflection fluorescence microscopy with millisecond time resolution, we were able to determine the optimal number of SNARE complexes needed for fast fusion. Here, we utilize this assay in combination wi

SUBMITTER: Kiessling V 

PROVIDER: S-EPMC3000493 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature