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Vol. 58, Issue 5, 954-966, November 2000
2-Adrenoceptor Coupling to
Gs-, Gi-, and Gq-Proteins
Howard Hughes Medical Institute, Stanford University Medical
School, Stanford, California; and Department of Pharmacology and
Toxicology, The University of Kansas, Lawrence, Kansas
The 
2-adrenoceptor (2AR) couples to the
G-protein Gs to activate adenylyl cyclase. Intriguingly,
several studies have demonstrated that the 2AR can also
interact with G-proteins of the Gi- and Gq-family. To assess the efficiency of 2AR
interaction with various G-protein 
-subunits (Gx
), we
expressed fusion proteins of the 2AR with the long
(GsL) and short
(GsS) splice variants of Gs,
the Gi-proteins Gi2 and
Gi3, and the Gq-proteins
Gq and G16 in Sf9 cells. Fusion proteins provide a rigorous approach for comparing the coupling of a given receptor to Gx because of the defined 1:1 stoichiometry
of receptor and G-protein and the efficient coupling. Here, we show that the 2AR couples to Gs-,
Gi-, and Gq-proteins as assessed by ternary
complex formation and ligand-regulated guanosine
5'-O-(3-thiotriphosphate) (GTP
S) binding. The
combined analysis of ternary complex formation, GTPS binding,
agonist efficacies, and agonist potencies revealed substantial
differences in the interaction of the 2AR with the various classes of G-proteins. Comparison of the coupling of the 2AR and formyl peptide receptor to
Gi2 revealed receptor-specific differences
in the kinetics of GTPS binding. We also detected highly efficient
stimulation of GTPS dissociation from
GsL, but not from Gq and
G16, by a 2AR agonist. Moreover, we show
that the 1:1 stoichiometry of receptor to G-protein in fusion proteins
reflects the in vivo stoichiometry of receptor/G-protein coupling more
closely than was previously assumed. Collectively, our data show 1)
that the 2AR couples differentially to Gs-, Gi-, and Gq-proteins, 2) that there is
ligand-specific coupling of the 2AR to G-proteins, 3)
that receptor-specific G-protein conformational states may exist, and
4) that nucleotide dissociation is an important mechanism for G-protein deactivation.
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