A fully automated [35S]GTPgammaS scintillation proximity assay for the high-throughput screening of Gi-linked G protein-coupled receptors.
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.
Ferrer
Marc
M
Department of Automated Biotechnology, Merck Research Laboratories, North Wales, Pennsylvania, USA.
Kolodin
Garrett D
GD
Zuck
Paul
P
Peltier
Richard
R
Berry
Kurtis
K
Mandala
Suzanne M
SM
Rosen
Hugh
H
Ota
Hisashi
H
Ozaki
Satoshi
S
Inglese
James
J
Strulovici
Berta
B
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
D043562:Receptors, G-Protein-Coupled; D013462:Sulfur Radioisotopes; D016244:Guanosine 5'-O-(3-Thiotriphosphate)