title: Systematic protein-protein interaction mapping for clinically relevant human GPCRs.
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.
authors.affiliation: Donnelly Centre, University of Toronto, Toronto, ON, Canada.
authors.forename: Kate
authors.initials: K
authors.lastname: Sokolina
authors.affiliation: Donnelly Centre, University of Toronto, Toronto, ON, Canada.
authors.forename: Saranya
authors.initials: S
authors.lastname: Kittanakom
authors.affiliation: Donnelly Centre, University of Toronto, Toronto, ON, Canada.
authors.forename: Jamie
authors.initials: J
authors.lastname: Snider
authors.affiliation: Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, Canada.
authors.forename: Max
authors.initials: M
authors.lastname: Kotlyar
authors.affiliation: Inserm, U1016, Institut Cochin, Paris, France.|CNRS UMR 8104, Paris, France.|Sorbonne Paris Cité, University of Paris Descartes, Paris, France.|UMR CNRS 7369 Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Université de Reims Champagne Ardenne (URCA), UFR Sciences Exactes et Naturelles, Reims, France.
authors.forename: Pascal
authors.identifier: 0000-0003-2167-4808
authors.initials: P
authors.lastname: Maurice
authors.affiliation: Unitat de Farmacologia, Departament de Patologia i Terapèutica Experimental, Facultat de Medicina, IDIBELL, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain.|Institut de Neurociències, Universitat de Barcelona, Barcelona, Spain.
authors.forename: Jorge
authors.identifier: 0000-0003-1711-8075
authors.initials: J
authors.lastname: Gandía
authors.affiliation: Inserm, U1016, Institut Cochin, Paris, France.|CNRS UMR 8104, Paris, France.|Sorbonne Paris Cité, University of Paris Descartes, Paris, France.
authors.forename: Abla
authors.initials: A
authors.lastname: Benleulmi-Chaachoua
authors.affiliation: Inserm, U1016, Institut Cochin, Paris, France.|CNRS UMR 8104, Paris, France.|Sorbonne Paris Cité, University of Paris Descartes, Paris, France.
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Tadagaki
authors.affiliation: Inserm, U1016, Institut Cochin, Paris, France.|CNRS UMR 8104, Paris, France.|Sorbonne Paris Cité, University of Paris Descartes, Paris, France.
authors.forename: Atsuro
authors.initials: A
authors.lastname: Oishi
authors.affiliation: Donnelly Centre, University of Toronto, Toronto, ON, Canada.
authors.forename: Victoria
authors.initials: V
authors.lastname: Wong
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, SK, Canada.
authors.forename: Ramy H
authors.initials: RH
authors.lastname: Malty
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, SK, Canada.
authors.forename: Viktor
authors.initials: V
authors.lastname: Deineko
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, SK, Canada.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Aoki
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, SK, Canada.
authors.forename: Shahreen
authors.initials: S
authors.lastname: Amin
authors.affiliation: Donnelly Centre, University of Toronto, Toronto, ON, Canada.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Yao
authors.affiliation: Unitat de Farmacologia, Departament de Patologia i Terapèutica Experimental, Facultat de Medicina, IDIBELL, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain.|Institut de Neurociències, Universitat de Barcelona, Barcelona, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Morató
authors.affiliation: Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, Canada.
authors.forename: David
authors.initials: D
authors.lastname: Otasek
authors.affiliation: Department of Biochemistry, Institute for Research in Immunology & Cancer, Université de Montréal, Montréal, QC, Canada.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kobayashi
authors.affiliation: Donnelly Centre, University of Toronto, Toronto, ON, Canada.
authors.forename: Javier
authors.initials: J
authors.lastname: Menendez
authors.affiliation: Dualsystems Biotech AG, Schlieren, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Auerbach
authors.affiliation: Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy and Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, ON, Canada.
authors.forename: Stephane
authors.initials: S
authors.lastname: Angers
authors.affiliation: Department of Computing, University College London, London, UK.
authors.forename: Natasa
authors.initials: N
authors.lastname: Pržulj
authors.affiliation: Department of Biochemistry, Institute for Research in Immunology & Cancer, Université de Montréal, Montréal, QC, Canada.
authors.forename: Michel
authors.initials: M
authors.lastname: Bouvier
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, SK, Canada.
authors.forename: Mohan
authors.initials: M
authors.lastname: Babu
authors.affiliation: Unitat de Farmacologia, Departament de Patologia i Terapèutica Experimental, Facultat de Medicina, IDIBELL, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain.|Institut de Neurociències, Universitat de Barcelona, Barcelona, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Ciruela
authors.affiliation: Inserm, U1016, Institut Cochin, Paris, France.|CNRS UMR 8104, Paris, France.|Sorbonne Paris Cité, University of Paris Descartes, Paris, France.
authors.forename: Ralf
authors.initials: R
authors.lastname: Jockers
authors.affiliation: Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, Canada.|Departments of Medical Biophysics and Computer Science, University of Toronto, Toronto, ON, Canada.|Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava, Slovakia.
authors.forename: Igor
authors.identifier: 0000-0002-2507-946X
authors.initials: I
authors.lastname: Jurisica
authors.affiliation: Donnelly Centre, University of Toronto, Toronto, ON, Canada igor.stagljar@utoronto.ca.|Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.|Department of Biochemistry, University of Toronto, Toronto, ON, Canada.
authors.forename: Igor
authors.identifier: 0000-0002-5260-3327
authors.initials: I
authors.lastname: Stagljar
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D043705:Receptor, Adenosine A2A / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D044422:Receptors, Serotonin, 5-HT4 / Q000378:metabolism; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
chemical_list: C480827:GPR37 receptor, human; D043705:Receptor, Adenosine A2A; D043562:Receptors, G-Protein-Coupled; D044422:Receptors, Serotonin, 5-HT4
keywords: G‐protein‐coupled receptors; high‐throughput screening; integrative computational biology; interactome; protein–protein interactions; split‐ubiquitin membrane yeast two‐hybrid assay