In Vitro Reconstitution Studies of SNAREs and Their Regulators Mediating GLUT4 Vesicle Fusion

AbstractThe GLUT4 vesicle fusion is mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) and a variety of regulatory proteins. For example, synip and tomosyn negatively regulate GLUT4 SNARE-mediated membrane fusion. Here we describe in vitro reconstituted assays to determine the molecular mechanisms of SNAREs, synip, and tomosyn. These methods can also be extended to the studies of other types of membrane fusion events.Key wordsGLUT4SynipTomosynVesicle fusionLiposome reconstitutionLipid mixingContent mixingSNARE assemblyDocking