GPCR-MAPS: high-resolution functional and allosteric mapping of G-protein-coupled receptor activation functions and bias

Abstract G-protein-coupled receptors (GPCRs) are important drug targets. Approaches targeting allosteric modulation and biased signaling, as opposed to the orthosteric pocket, can improve safety profiles of therapeutics. However, rational design of precision therapies is limited by the lack of high-resolution quantitative functional annotation of GPCR structures. Here we present GPCR-MAPS—a fast and general approach to build complete, high-resolution, multimodal functional and allosteric maps of GPCRs. GCPR-MAPS quantifies direct recruitment of proteins and deconvolves the substantial effects of amino acid substitutions on receptor expression. We applied GPCR-MAPS to the β 2 adrenergic receptor to generate >7,500 dose–response curves and reveal a modular receptor architecture, with a core activation network surrounded by residues controlling quantitative parameters and bias. We used this information to identify multiple allosteric surface pockets. The application of this approach across the superfamily of GPCRs could unlock deep understanding of these receptors and enable rational design of precision GPCR therapeutics.