High throughput functional assay for G-protein coupled receptors using a rap-ras chimeric protein

摘要

Disclosed is a method of determining whether a substance is a ligand for a G-protein coupled receptor (GPCR), comprising: (a) providing a cell culture which comprises cells comprising a first nucleic acid which encodes a GPCR which increases or decreases cellular proliferation through a GPCR cascade in response to a ligand, a second recombinant nucleic acid which encodes a marker indicative of the extent of cellular proliferation, and a third recombinant nucleic acid encoding a Ras effector domain and a Rap input domain operably coupled to said Ras effector domain; (b) contacting said cell culture with a test compound; and (c) determining the level of cellular proliferation