Core engagement with β-arrestin is dispensable for agonist-induced vasopressin receptor endocytosis and ERK activation.

G protein-coupled receptors (GPCRs) exhibit highly conserved activation and signaling mechanisms by which agonist stimulation leads to coupling of heterotrimeric G proteins and generation of second messenger response. This is followed by receptor phosphorylation, primarily in the carboxyl terminus but also in the cytoplasmic loops, and subsequent binding of arrestins. GPCRs typically recruit arrestins through two different sets of interactions, one involving phosphorylated receptor tail and the other mediated by the receptor core. The engagement of both set of interactions (tail and core) is generally believed to be necessary for arrestin-dependent functional outcomes such as receptor desensitization, endocytosis, and G protein-independent signaling. Here we demonstrate that a vasopressin receptor (VR) mutant with truncated third intracellular loop (VR) can interact with β-arrestin 1 (βarr1) only through the phosphorylated tail without engaging the core interaction. Of interest, such a partially engaged VR-βarr1 complex can efficiently interact with clathrin terminal domain and ERK2 MAPK in vitro. Furthermore, this core interaction-deficient VR mutant exhibits efficient endocytosis and ERK activation upon agonist stimulation. Our data suggest that core interaction with βarr is dispensable for VR endocytosis and ERK activation and therefore provide novel insights into refining the current understanding of functional requirements in biphasic GPCR-βarr interaction.