RT Journal Article SR Electronic T1 Stereochemistry of an Agonist Determines Coupling Preference of β2-Adrenoceptor to Different G Proteins in Cardiomyocytes JF Molecular Pharmacology JO Mol Pharmacol FD American Society for Pharmacology and Experimental Therapeutics SP 158 OP 165 DO 10.1124/mol.108.051078 VO 75 IS 1 A1 Woo, Anthony Yiu-Ho A1 Wang, Tian-Bing A1 Zeng, Xiaokun A1 Zhu, Weizhong A1 Abernethy, Darrell R. A1 Wainer, Irving W. A1 Xiao, Rui-Ping YR 2009 UL http://molpharm.aspetjournals.org/content/75/1/158.abstract AB A fundamental question regarding receptor-G protein interaction is whether different agonists can lead a receptor to different intracellular signaling pathways. Our previous studies have demonstrated that although most β2-adrenoceptor agonists activate both Gs and Gi proteins, fenoterol, a full agonist of β2-adrenoceptor, selectively activates Gs protein. Fenoterol contains two chiral centers and may exist as four stereoisomers. We have synthesized a series of stereoisomers of fenoterol and its derivatives and characterized their receptor binding and pharmacological properties. We tested the hypothesis that the stereochemistry of an agonist determines selectivity of receptor coupling to different G protein(s). We found that the R,R isomers of fenoterol and methoxyfenoterol exhibited more potent effects to increase cardiomyocyte contraction than their S,R isomers. It is noteworthy that although (R,R)-fenoterol and (R,R)-methoxyfenoterol preferentially activate Gs signaling, their S,R isomers were able to activate both Gs and Gi proteins as evidenced by the robust pertussis toxin sensitivities of their effects on cardiomyocyte contraction and on phosphorylation of extracellular signal-regulated kinase 1/2. The differential G protein selectivities of the fenoterol stereoisomers were further confirmed by photoaffinity labeling studies on Gs,Gi2, and Gi3 proteins. The inefficient Gi signaling with the R,R isomers is not caused by the inability of the R,R isomers to trigger the protein kinase A (PKA)-mediated phosphorylation of the β2-adrenoceptor, because the R,R isomers also markedly increased phosphorylation of the receptor at serine 262 by PKA. We conclude that in addition to receptor subtype and phosphorylation status, the stereochemistry of a given agonist plays an important role in determining receptor-G protein selectivity and downstream signaling events. U.S. Government work not protected by U.S. copyright