Conformational dynamics of the active state of β-arrestin 1

Abstract

{beta}-arrestins ({beta}arr) regulate the signaling and trafficking of G protein-coupled receptors (GPCRs) in numerous physiological processes and have been implicated in various diseases. Structural and kinetic insights into how ligand-mediated GPCR activation drives {beta}arr coupling and activation remain limited, with the binding mechanism of the phosphorylated GPCR C-terminal tails, such as that of the vasopressin receptor-2 (V2Rpp), and the conformation of the entire {beta}arr tail in the active state still unknown. Here, we simulated both the basal and V2Rpp-bound states of {beta}arr1 with temperature replica-exchange molecular dynamics (TREMD) simulations to probe the activation mechanism of {beta}arr1. Compared to conventional MD, our TREMD simulations, employing an unprecedented 200-replica setup, significantly broadened conformational sampling while preserving the basal state. Our analysis showed that, without the bound Fab30 antibody fragment, the main body of V2Rpp-bound {beta}arr1 tended to transition toward the basal conformation; however, binding of V2Rpp in the N-domain groove allosterically oriented the finger loop to point upward for core engagement with a GPCR. Furthermore, V2Rpp dissociation events suggest that its binding involves a sliding movement along the N-domain groove, during which its phosphorylated residues p3 and p4 transiently occupy the S5 site to facilitate repositioning of p5 into the S5 site, thereby triggering a zippering process of p1 to p3. The dynamic 62-residue {beta}arr1 tail explored a vast conformational space, forming transient secondary structures, and could favorably anchor on the main bodys back side and within the central crest crevice. These findings elucidate key mechanistic steps underlying {beta}arr1 activation.