title: Oligomerization of sweet and bitter taste receptors.
abstract: The superfamily of G protein-coupled receptors (GPCRs) mediates numerous physiological processes, including neurotransmission, cell differentiation and metabolism, and sensory perception. In recent years, it became evident that these receptors might function not only as monomeric receptors but also as homo- or heteromeric receptor complexes. The family of TAS1R taste receptors are prominent examples of GPCR dimerization as they act as obligate functional heteromers: TAS1R1 and TAS1R3 combine to form an umami taste receptor, while the combination of TAS1R2 and TAS1R3 is a sweet taste receptor. So far, TAS2Rs, a second family of ~25 taste receptors in humans that mediates responses to bitter compounds, have been shown to function on their own, but if they do so as receptor monomers or as homomeric receptors still remains unknown. Using two different experimental approaches, we have recently shown that TAS2Rs can indeed form both homomeric and heteromeric receptor complexes. The employed techniques, coimmunoprecipitations and bioluminescence resonance energy transfer (BRET), are based on different principles and complement each other well and therefore provided compelling evidences for TAS2R oligomerization. Furthermore, we have adapted the protocols to include a number of controls and for higher throughput to accommodate the investigation of a large number of receptors and receptor combinations. Here, we present the protocols in detail.
authors.affiliation: Axxam SpA, Bresso (Milan), Italy.
authors.forename: Christina
authors.initials: C
authors.lastname: Kuhn
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Meyerhof
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods*; 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*; D010957:Plasmids; D011485:Protein Binding; D055503:Protein Multimerization; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013649:Taste / Q000502:physiology; D014162:Transfection; D000098706:Taste Receptors, Type 2
chemical_list: D043562:Receptors, G-Protein-Coupled; C431926:taste receptors, type 1; D000098706:Taste Receptors, Type 2; D049452:Green Fluorescent Proteins; D049410:Luciferases, Renilla
keywords: BRET; Bitter taste receptors; CoIP; Gustation; Oligomerization; Sweet taste receptor; T1R; T2R; TAS1R; TAS2R