The GTPase-activating Protein RGS4 Stabilizes the Transition State for Nucleotide Hydrolysis

RGS proteins constitute a newly appreciated group of negative regulators of G protein signaling. Discovered by genetic screens in yeast, worms, and other organisms, two mammalian RGS proteins, RGS4 and GAIP, act as GTPase-activating proteins for members of the Gi family of G protein α subunits. We have purified recombinant RGS4 to homogeneity and demonstrate that it acts catalytically to stimulate GTP hydrolysis by Gi proteins. Furthermore, RGS4 stabilizes the transition state for GTP hydrolysis, as evidenced by its high affinity for the GDP-AlF4−-bound forms of Goα and Giα and its relatively low affinity for the GTPγS- and GDP-bound forms of these proteins. Consequently, RGS4 is most likely not a downstream effector for activated Gα subunits. All members of the Gi subfamily of proteins tested are substrates for RGS4 (including Gtα and Gzα); the protein has lower affinity for Gqα, and it does not stimulate the GTPase activity of Gsα or G12α.