GPCR screening via ERK 1/2: a novel platform for screening G protein-coupled receptors.
Discovery of novel agonists and antagonists for G protein-coupled receptors (GPCRs) relies heavily on cell-based assays because determination of functional consequences of receptor engagement is often desirable. Currently, there are several key parameters measured to achieve this, including mobilization of intracellular Ca2+ and formation of cyclic adenosine monophosphate or inositol triphosphate. However, no single assay platform is suitable for all situations, and all of the assays have limitations. The authors have developed a new high-throughput homogeneous assay platform for GPCR discovery as an alternative to current assays, which employs detection of phosphorylation of the key signaling molecule p42/44 MAP kinase (ERK 1/2). The authors show that ERK 1/2 is consistently activated in cells stimulated by Gq-coupled GPCRs and provides a new high-throughput platform for screening GPCR drug candidates. The activation of ERK 1/2 in Gq-coupled GPCR systems generates comparable pharmacological data for receptor agonist and antagonist data obtained by other GPCR activation measurement techniques.
Osmond
Ronald I W
RI
TGR BioSciences Pty Ltd, 31 Dalgleish Street, Thebarton, SA 5031, Australia.
Sheehan
Antony
A
Borowicz
Romana
R
Barnett
Emma
E
Harvey
Georgina
G
Turner
Cheryl
C
Brown
Andrea
A
Crouch
Michael F
MF
Dyer
Anthony R
AR
D000818:Animals; D019556:COS Cells; D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D002522:Chlorocebus aethiops; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics / Q000378:metabolism*; D009496:Neurotensin / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000032:analysis* / Q000037:antagonists & inhibitors; D014162:Transfection
D043562:Receptors, G-Protein-Coupled; D009496:Neurotensin; D002217:Carbachol; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3