Towards reconstitution of membrane fusion mediated by SNAREs and other synaptic proteins

Critical Reviews in Biochemistry and Molecular Biology
Axel T BrungerJiajie Diao

Abstract

Proteoliposomes have been widely used for in vitro studies of membrane fusion mediated by synaptic proteins. Initially, such studies were made with large unsynchronized ensembles of vesicles. Such ensemble assays limited the insights into the SNARE-mediated fusion mechanism that could be obtained from them. Single particle microscopy experiments can alleviate many of these limitations but they pose significant technical challenges. Here we summarize various approaches that have enabled studies of fusion mediated by SNAREs and other synaptic proteins at a single-particle level. Currently available methods are described and their advantages and limitations are discussed.

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Methods Mentioned

BETA
FRET
pull-down
single particle microscopy
confocal microscopy
fluorescence
proteolysis assay

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