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Systematic protein–protein interaction mapping for clinically relevant human GPCRs |
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| Authors: | Max Kotlyar Pascal Maurice Jorge Gandía Abla Benleulmi‐Chaachoua Kenjiro Tadagaki Atsuro Oishi Victoria Wong Ramy H Malty Viktor Deineko Hiroyuki Aoki Shahreen Amin Zhong Yao Xavier Morató David Otasek Hiroyuki Kobayashi Javier Menendez Daniel Auerbach Stephane Angers Natasa Pr?ulj Michel Bouvier Mohan Babu Francisco Ciruela Ralf Jockers Igor Jurisica Igor Stagljar |
| Institution: | 1. Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, Canada;2. Inserm, U1016, Institut Cochin, Paris, France;3. CNRS UMR 8104, Paris, France;4. Sorbonne Paris Cité, University of Paris Descartes, Paris, France;5. UMR CNRS 7369 Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Université de Reims Champagne Ardenne (URCA), UFR Sciences Exactes et Naturelles, Reims, France;6. Unitat de Farmacologia, Departament de Patologia i Terapèutica Experimental, Facultat de Medicina, IDIBELL, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain;7. Institut de Neurociències, Universitat de Barcelona, Barcelona, Spain;8. Donnelly Centre, University of Toronto, Toronto, ON, Canada;9. Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, SK, Canada;10. Department of Biochemistry, Institute for Research in Immunology & Cancer, Université de Montréal, Montréal, QC, Canada;11. Dualsystems Biotech AG, Schlieren, Switzerland;12. Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy and Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, ON, Canada;13. Department of Computing, University College London, London, UK;14. Departments of Medical Biophysics and Computer Science, University of Toronto, Toronto, ON, Canada;15. Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava, Slovakia;16. Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada;17. Department of Biochemistry, University of Toronto, Toronto, ON, Canada |
| 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. |
| Keywords: | G‐protein‐coupled receptors high‐throughput screening integrative computational biology interactome protein– protein interactions split‐ubiquitin membrane yeast two‐hybrid assay |