RT Journal Article SR Electronic T1 Estimation of Agonist Activity at G Protein-Coupled Receptors: Analysis of M2 Muscarinic Receptor Signaling through Gi/o,Gs, and G15 JF Journal of Pharmacology and Experimental Therapeutics JO J Pharmacol Exp Ther FD American Society for Pharmacology and Experimental Therapeutics SP 1193 OP 1207 DO 10.1124/jpet.107.120857 VO 321 IS 3 A1 Michael T. Griffin A1 Katherine W. Figueroa A1 Sarah Liller A1 Frederick J. Ehlert YR 2007 UL http://jpet.aspetjournals.org/content/321/3/1193.abstract AB We developed novel methods for analyzing the concentration-response curve of an agonist to estimate the product of observed affinity and intrinsic efficacy, expressed relative to that of a standard agonist. This parameter, termed intrinsic relative activity (RAi), is most applicable for the analysis of responses at G protein-coupled receptors. RAi is equivalent to the potency ratios that agonists would exhibit in a hypothetical, highly sensitive assay in which all agonists behave as full agonists, even those with little intrinsic efficacy. We investigated muscarinic responses at the M2 receptor, including stimulation of phosphoinositide hydrolysis through Gα15 in HEK 293T cells, inhibition of cAMP accumulation through Gi in Chinese hamster ovary (CHO) cells, and stimulation of cAMP accumulation through Gs in CHO cells treated with pertussis toxin. The RAi values of carbachol, oxotremorine-M, and the enantiomers of aceclidine were approximately the same in the three assay systems. In contrast, the activity of 4-[[N-[3-chlorophenyl]carbamoy]oxy-2-butynyl]trimethylammonium chloride (McN-A-343) was ∼10-fold greater at M2 receptors coupled to Gα15 in HEK 293T cells compared with M2 receptors coupled to Gi in the same cells or in CHO cells. Our results show that the RAi estimate is a useful measure for quantifying agonist activity across different assay systems and for detecting agonist directed signaling. The American Society for Pharmacology and Experimental Therapeutics