title: Application of BRET to monitor ligand binding to GPCRs.
abstract: Bioluminescence resonance energy transfer (BRET) is a well-established method for investigating protein-protein interactions. Here we present a BRET approach to monitor ligand binding to G protein-coupled receptors (GPCRs) on the surface of living cells made possible by the use of fluorescent ligands in combination with a bioluminescent protein (NanoLuc) that can be readily expressed on the N terminus of GPCRs.
authors.affiliation: Cell Signalling Research Group, School of Life Sciences, University of Nottingham Medical School, Nottingham, United Kingdom.
authors.forename: Leigh A
authors.initials: LA
authors.lastname: Stoddart
authors.affiliation: Molecular Endocrinology and Pharmacology, Harry Perkins Institute of Medical Research, Nedlands, Western Australia, Australia.|Centre for Medical Research, The University of Western Australia, Crawley, Western Australia, Australia.
authors.forename: Elizabeth K M
authors.initials: EKM
authors.lastname: Johnstone
authors.affiliation: Cell Signalling Research Group, School of Life Sciences, University of Nottingham Medical School, Nottingham, United Kingdom.
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Wheal
authors.affiliation: Cell Signalling Research Group, School of Life Sciences, University of Nottingham Medical School, Nottingham, United Kingdom.
authors.forename: Joëlle
authors.initials: J
authors.lastname: Goulding
authors.affiliation: Promega Corporation, Madison, Wisconsin, United States.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Robers
authors.affiliation: Promega Corporation, Madison, Wisconsin, United States.
authors.forename: Thomas
authors.initials: T
authors.lastname: Machleidt
authors.affiliation: Promega Corporation, Madison, Wisconsin, United States.
authors.forename: Keith V
authors.initials: KV
authors.lastname: Wood
authors.affiliation: Cell Signalling Research Group, School of Life Sciences, University of Nottingham Medical School, Nottingham, United Kingdom.|Molecular Endocrinology and Pharmacology, Harry Perkins Institute of Medical Research, Nedlands, Western Australia, Australia.|Centre for Medical Research, The University of Western Australia, Crawley, Western Australia, Australia.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Hill
authors.affiliation: Molecular Endocrinology and Pharmacology, Harry Perkins Institute of Medical Research, Nedlands, Western Australia, Australia.|Centre for Medical Research, The University of Western Australia, Crawley, Western Australia, Australia.
authors.forename: Kevin D G
authors.initials: KDG
authors.lastname: Pfleger
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods*; D005453:Fluorescence; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*
chemical_list: D008024:Ligands; D018343:Receptors, Adrenergic, beta-2; D043562:Receptors, G-Protein-Coupled