title: A fully automated [35S]GTPgammaS scintillation proximity assay for the high-throughput screening of Gi-linked G protein-coupled receptors.
abstract: The diversity of physiological functions mediated by the GPCR superfamily provides a rich source of molecular targets for drug discovery programs. Consequently, a variety of assays have been designed to identify lead molecules based on ligand binding or receptor function. In one of these, the binding of [(35)S]GTPgammaS, a nonhydrolyzable analogue of GTP, to receptor-activated G-protein alpha subunits represents a unique functional assay for GPCRs and is well suited for use with automated HTS. Here we compare [(35)S]GTPgammaS scintillation proximity binding assays for two different G(i)-coupled GPCRs, and describe their implementation with automated high-throughput systems.
authors.affiliation: Department of Automated Biotechnology, Merck Research Laboratories, North Wales, Pennsylvania, USA.
authors.forename: Marc
authors.initials: M
authors.lastname: Ferrer
authors.forename: Garrett D
authors.initials: GD
authors.lastname: Kolodin
authors.forename: Paul
authors.initials: P
authors.lastname: Zuck
authors.forename: Richard
authors.initials: R
authors.lastname: Peltier
authors.forename: Kurtis
authors.initials: K
authors.lastname: Berry
authors.forename: Suzanne M
authors.initials: SM
authors.lastname: Mandala
authors.forename: Hugh
authors.initials: H
authors.lastname: Rosen
authors.forename: Hisashi
authors.initials: H
authors.lastname: Ota
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ozaki
authors.forename: James
authors.initials: J
authors.lastname: Inglese
authors.forename: Berta
authors.initials: B
authors.lastname: Strulovici
mesh_terms: D001331:Automation / Q000379:methods*; D001681:Biological Assay / Q000379:methods; D020650:Combinatorial Chemistry Techniques / Q000379:methods; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000032:analysis* / Q000378:metabolism; D011869:Radioligand Assay / Q000379:methods; D043562:Receptors, G-Protein-Coupled / Q000032:analysis*; D012588:Scintillation Counting / Q000379:methods*; D013462:Sulfur Radioisotopes / Q000378:metabolism
chemical_list: D043562:Receptors, G-Protein-Coupled; D013462:Sulfur Radioisotopes; D016244:Guanosine 5'-O-(3-Thiotriphosphate)