Detection of receptor ligands by monitoring selective stabilization of a Renilla luciferase-tagged, constitutively active mutant, G-protein-coupled receptor

摘要

The wild-typeβ_2-adrenoceptor and a constitutively active mutant of this receptor were C- terminally tagged with luciferase from the pansy Renilla reniformis. C-terminal addition of Renilla luciferase did not substantially alter the levels of expression of Either form of the receptor, the elevated constitutive activity of the mutantβ_2-adrenoceptor nor the Capacity of isoprenaline to elevate cyclic AMP levels in intact cells expressing these constructs. Treatment of cells expressing constitutively active mutantβ_2-adrenoceptor-renilla luciferase with Antagonist/inverse agonist ligands resulted in upregulation of levels of this polypeptide which could Be monitored by the elevated luciferase activity.