title: Time-resolved FRET strategy to screen GPCR ligand library.
abstract: Screening chemical libraries to find specific drugs for G protein-coupled receptors is still of major interest. Indeed, because of their major roles in all physiological functions, G protein-coupled receptors remain major targets for drug development programs. Currently, interest in GPCRs as drug targets has been boosted by the discovery of biased ligands, thus allowing the development of drugs not only specific for one target but also for the specific signaling cascade expected to have the therapeutic effect. Such molecules are then expected to display fewer side effects. To reach such a goal, there is much interest in novel, efficient, simple, and direct screening assays that may help identify any drugs interacting with the target, these being then analyzed for their biased activity. Here, we present an efficient strategy to screen ligands on their binding properties. The method described is based on time-resolved FRET between a receptor and a ligand. This method has already been used to develop new assays called Tag-lite(®) binding assays for numerous G protein-coupled receptors, proving its broad application and its power.
authors.affiliation: Institut de Génomique Fonctionnelle, CNRS, UMR 5203, 141 Rue de la Cardonille, 34094, Montpellier Cedex 5, France.
authors.forename: Nadia
authors.initials: N
authors.lastname: Oueslati
authors.forename: Candide
authors.initials: C
authors.lastname: Hounsou
authors.forename: Abderazak
authors.initials: A
authors.lastname: Belhocine
authors.forename: Thieric
authors.initials: T
authors.lastname: Rodriguez
authors.forename: Elodie
authors.initials: E
authors.lastname: Dupuis
authors.forename: Jurriaan M
authors.initials: JM
authors.lastname: Zwier
authors.forename: Eric
authors.initials: E
authors.lastname: Trinquet
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Pin
authors.forename: Thierry
authors.initials: T
authors.lastname: Durroux
mesh_terms: D001665:Binding Sites; D056831:Coordination Complexes / Q000737:chemistry; D015195:Drug Design; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D015870:Gene Expression; D006146:Guanidines / Q000737:chemistry; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D007700:Kinetics; D008024:Ligands; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013194:Staining and Labeling / Q000379:methods*; D013725:Terbium / Q000737:chemistry
chemical_list: D056831:Coordination Complexes; D005456:Fluorescent Dyes; D006146:Guanidines; D008024:Ligands; D043562:Receptors, G-Protein-Coupled; D054852:Small Molecule Libraries; D013725:Terbium; C117872:benzylguanidine; D019853:O(6)-Methylguanine-DNA Methyltransferase