Systematic protein–protein interaction mapping for clinically relevant human GPCR s

Abstract

G‐protein‐coupled receptors ( GPCR s) are the largest family of integral membrane receptors with key roles in regulating signaling pathways targeted by therapeutics, but are difficult to study using existing proteomics technologies due to their complex biochemical features. To obtain a global view of GPCR ‐mediated signaling and to identify novel components of their pathways, we used a modified membrane yeast two‐hybrid ( MYTH ) approach and identified interacting partners for 48 selected full‐length human ligand‐unoccupied GPCR s in their native membrane environment. The resulting GPCR interactome connects 686 proteins by 987 unique interactions, including 299 membrane proteins involved in a diverse range of cellular functions. To demonstrate the biological relevance of the GPCR interactome, we validated novel interactions of the GPR 37, serotonin 5‐ HT 4d, and adenosine ADORA 2A receptors. Our data represent the first large‐scale interactome mapping for human GPCR s and provide a valuable resource for the analysis of signaling pathways involving this druggable family of integral membrane proteins. image The complete interactome of 48 disease‐associated, full‐length human G‐coupled protein receptors ( GPCR s) is mapped in the native environment of the cellular membrane, using the membrane yeast two‐hybrid ( MYTH ) technology. MYTH is used to identify interacting partners for 48 human GPCRS, resulting in an interactome containing 686 proteins and 987 unique interactions. Bioinformatics analyses of the GPCR interactome indicate enrichment for diverse pathways, diseases, molecular functions, biological processes, domains, and drug targets. Orthogonal analyses using co‐immunoprecipitation and BRET validate a subset of the identified interactions. Functional characterization of novel GPCR interactions identifies potential roles in neurobiological processes.