G-protein-coupled receptor interaction with small GTPases

In addition to heterotrimeric G-proteins, Ras-like small GTPases are also involved in regulating physiological functions of the G-protein-coupled receptor (GPCR) superfamily. In particular, Rab and ARF GTPases function either as "traffic cops" to coordinate receptor targeting to specific locations or as "signal transducers" to directly mediate receptor signal propagation. As revealed in protein-protein interaction assays, GPCRs may use specific motifs to physically interact with small GTPases, providing important insights into the underlying molecular mechanisms. In this chapter, we describe coimmunoprecipitation and GST fusion protein pull-down approaches to study the GPCR-small GTPase interaction, by focusing on the interaction of α_2B- and β₂-adrenegic receptors with the small GTPases Rab8 and ARF1.