Methods to Investigate the beta-Arrestin-Mediated Control of ARF6 Activation to Regulate Trafficking and Actin Cytoskeleton Remodeling

ADP-ribosylation factors (ARF) are GTPases that act to control the activation of numerous signaling events and cellular responses. The ARF6 isoform, present at the plasma membrane, can be activated by the angiotensin II type 1 receptor (AT(1)R), a process dependent upon beta-arrestin recruitment to the activated receptor. Here, we describe classical methods used to assess beta-arrestin-dependent activation of ARF6 following agonist stimulation of cells. In addition, because ARF6 and beta-arrestin can form a complex, we describe the procedures used to detect the interaction of beta-arrestin with this GTPase.