Decoding Angiotensin II Type 1 Receptor Allosteric Communication to Gq and β-arrestin

The allosteric communication between the agonist binding site and the G protein or {beta}-arrestin coupling sites in G protein-coupled receptors (GPCRs) play an important role in determining ligand efficacy towards these two signaling pathways and hence the ligand bias. Knowledge of the amino acid residue networks involved in the allosteric communication will aid understanding GPCR signaling and the design of biased ligands. Angiotensin II type I receptor (AT1R) is an ideal model GPCR to study the molecular basis of ligand bias as it has multiple {beta}-arrestin2 and Gq protein biased agonists as well as three-dimensional structures. Using Molecular Dynamics simulations, dynamic allostery analysis, and functional BRET assays, we identified a network of residues involved in allosteric communication from the angiotensin II binding site to the putative Gq coupling sites and another network to the {beta}-arrestin2 coupling sites, with 6 residues common to both pathways located in TM3, TM5 and TM6. Our findings unveil unique and common allosteric communication residue hubs for Gq and {beta}-arr2 coupling by AngII ligands and suggests that some of these residues can be targeted to design biased AT1R ligands. Finally, we show through analysis of the inter-residue distance distributions of the activation microswitches involved in class A GPCR activation for ten different agonists, that these microswitches behave like rheostats with different relative strengths of activation, which we speculate could modulate the relative efficacy of these agonists toward the two signaling pathways. Significance StatementKnowledge of the residues involved in allosteric communication from the ligand binding site to the G protein or {beta}-arrestin ({beta}-arr) coupling sites in GPCRs will aid in understanding their role in mediating ligand bias. Using a combination of molecular dynamics simulations and functional signaling assays we have identified a network of residues involved in allosteric communication from the Angiotensin II (Ang II) binding site to the Gq and {beta}-arr2 coupling sites in the Ang II type I receptor (AT1R). The residues in the allosteric network for {beta}-arr2 coupling are distributed across multiple structural regions of AT1R compared to Gq coupling. The residues in the two networks show conserved chemical properties across class A GPCRs, demonstrating the importance of allosteric communication in modulating ligand bias.