A convenient protocol for generating giant unilamellar vesicles containing SNARE proteins using electroformation.

ABSTRACT: Reconstitution of membrane proteins in artificial membranes is an essential prerequisite for functional studies that depend on the context of an intact membrane. While straight-forward protocols for reconstituting proteins in small unilamellar vesicles were developed many years ago, it is much more difficult to prepare large membranes containing membrane proteins at biologically relevant concentrations. Giant unilamellar vesicles (GUVs) represent a model system that is characterised by low curvature, controllable tension, and large surface that can be easily visualised with microscopy, but protein insertion is notoriously difficult. Here we describe a convenient method for efficient generation of GUVs containing functionally active SNARE proteins that govern exocytosis of synaptic vesicles

SUBMITTER: Witkowska A 

PROVIDER: S-EPMC6013450 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature