Binding of Rab3A to synaptic vesicles

Prenylated Rab GTPases cycle between membrane-bound and soluble forms. Membrane-bound GrDP-Rabs interact with GDP dissociation inhibitor (GDI), resulting in the dissociation of a Rab GDI complex, which in turn serves as a precursor for the membrane re-association of Rabs, We have now characterized the binding of Rab3A to synaptic vesicles in vitro using either purified completes or rat brain cytosol as source for GDI.Rab3A Binding of Rab3A results in the immediate release of GDI from the membrane. Furthermore, binding does not require the presence of additional guanine nucleotides (GDP or GTP) or of cytosolic factors. Although nucleotide exchange follows binding, binding is initially reversible, suggesting that binding of GDP-Rab3A and nucleotide exchange are separate and independent events. Comparison with the binding of Rab1B revealed that both Rab proteins bind preferentially to their respective resident membranes although some promiscuity was observable, Binding is saturable and involves a protease-sensitive binding site that is tightly associated with the vesicle membrane.