title: Biochemical assay of G protein-coupled receptor oligomerization: adenosine A1 and thromboxane A2 receptors form the novel functional hetero-oligomer.
abstract: G protein-coupled receptors (GPCRs) are classified into a family of seven transmembrane receptors. Receptor oligomerization may be the key to the expression and function of these receptors, for example, ligand binding, desensitization, membrane trafficking, and signaling. The accumulation of evidence that GPCRs form an oligomerization with a functional alternation may change the strategy for the discovery of novel drugs targeting GPCRs. Identification of the oligomer is essential to elucidate GPCR oligomerization. GPCR oligomerizations have been demonstrated using various biochemical approaches, which include the coimmunoprecipitation method, fluorescence resonance energy transfer assay, and bioluminescence RET assay. Thus, various assays are useful for the study of GPCR oligomerization, and we should choose the best method to match the purpose. We previously targeted adenosine A1 and thromboxane A2 (TP) receptors to form a functionally novel hetero-oligomer, since both receptors function in the same cells. This chapter describes the methods used to detect GPCR oligomerization and alterations in the signaling pathways, principally according to our findings on oligomerization between adenosine A1 and TPα receptors.
authors.affiliation: Department of Pharmacotherapy of Life-style Related Disease, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.
authors.forename: Natsumi
authors.initials: N
authors.lastname: Mizuno
authors.forename: Tokiko
authors.initials: T
authors.lastname: Suzuki
authors.forename: Yu
authors.initials: Y
authors.lastname: Kishimoto
authors.forename: Noriyasu
authors.initials: N
authors.lastname: Hirasawa
mesh_terms: D001667:Binding, Competitive; D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods*; D000242:Cyclic AMP / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D007700:Kinetics; D049410:Luciferases, Renilla / Q000235:genetics / Q000378:metabolism*; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D011485:Protein Binding; D055503:Protein Multimerization; D021381:Protein Transport; D043682:Receptor, Adenosine A1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D044244:Receptors, Thromboxane A2, Prostaglandin H2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection
chemical_list: D043682:Receptor, Adenosine A1; D044244:Receptors, Thromboxane A2, Prostaglandin H2; D049452:Green Fluorescent Proteins; D000242:Cyclic AMP; D049410:Luciferases, Renilla; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3
keywords: Adenosine A(1) receptor; BRET; Coimmunoprecipitation; GPCR; Oligomerization; TPα receptor