Systematic protein–protein interaction mapping for clinically relevant human GPCRs. Issue 3 (15th March 2017)

• Record Type: Journal Article
• Authors: Sokolina, Kate; Kittanakom, Saranya; Snider, Jamie; Kotlyar, Max; Maurice, Pascal; Gandía, Jorge; Benleulmi‐Chaachoua, Abla; Tadagaki, Kenjiro; Oishi, Atsuro; Wong, Victoria; Malty, Ramy H; Deineko, Viktor; Aoki, Hiroyuki; Amin, Shahreen; Yao, Zhong; Morató, Xavier; Otasek, David; Kobayashi, Hiroyuki; Menendez, Javier; Auerbach, Daniel; Angers, Stephane; Pržulj, Natasa; Bouvier, Michel; Babu, Mohan; Ciruela, Francisco; Jockers, Ralf; Jurisica, Igor; Stagljar, Igor
• Is Part Of: Molecular systems biology. Volume 13:Issue 3(2017:Mar.)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2017-03-15
• Subjects: G‐protein‐coupled receptors -- high‐throughput screening -- integrative computational biology -- interactome -- protein–protein interactions -- split‐ubiquitin membrane yeast two‐hybrid assay; Molecular biology -- Periodicals; Systems biology -- Periodicals; 572.8
• DOI: 10.15252/msb.20167430
• Languages: English
• ISSNs: 1744-4292

Abstract

Abstract: G‐protein‐coupled receptors (GPCRs) 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 GPCRs 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 GPR37, serotonin 5‐HT4d, and adenosine ADORA2A receptors. Our data represent the first large‐scale interactome mapping for human GPCRs and provide a valuable resource for the analysis of signaling pathways involving this druggable family of integral membrane proteins. Synopsis: The complete interactome of 48 disease‐associated, full‐length human G‐coupled protein receptors (GPCRs) 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. Abstract : The complete interactome of 48 disease‐associated, full‐length human G‐coupled protein receptors (GPCRs) is mapped in the native environment of the cellular membrane, using the membrane yeast two‐hybrid (MYTH) technology.