Interaction and functional analyses of human VPS34/p150 phosphatidylinositol 3-kinase complex with Rab7.

The Rab7 GTPase is a key regulator of late endocytic membrane transport and autophagy. Rab7 exerts temporal and spatial control over late endocytic membrane transport through interactions with various effector proteins. Among Rab7 effectors, the hVPS34/p150 phosphatidylinositol (PtdIns) 3-kinase complex serves to regulate late endosomal phosphatidylinositol signaling that is important for protein sorting and intraluminal vesicle sequestration. In this chapter, reagents and methods for the characterization of the interactions and regulation of the Rab7/hVPS34/p150 complex are described. Using these methods we demonstrate the requirement for activated Rab7 in the regulation of hVPS34/p150 PtdIns 3-kinase activity on late endosomes in vivo.