Analyses of the effects of Gq protein on the activated states of the muscarinic M3 receptor and the purinergic P2Y1 receptor

G protein-coupled receptors (GPCRs) cause various cellular responses through activating heterotrimeric G protein upon the agonist binding. The interaction with G protein has been suggested to stabilize the agonist-bound active conformation of GPCRs. We previously reported the effects of Gq protein on the stabilization of the active conformation of the muscarinic receptor type 1 (M1R), using a fluorescence resonance energy transfer (FRET) technique. In this study, we aimed at examining whether or not the binding of Gq protein affects the agonist-induced active conformation of receptors other than the M1R. For this purpose, functionally intact fluorescent receptors of the metabotropic purinergic receptor type 1 (P2Y1R) and muscarinic receptor type 3 (M3R) were constructed, by inserting junctional linkers between the short intracellular third loops (i3) and yellow fluorescent protein (YFP). The YFP-fused receptors also showed the agonist-induced increases in FRET from the cyan fluorescent protein (CFP) tethered with Gαq subunit, indicating that they interacted with Gq protein. The agonist-induced conformational changes of the receptors were detected as the agonist-induced decrease in FRET between YFP at the i3 and CFP at the C-tail. The FRET decrease of the M3R but not of the P2Y1R was enhanced by coexpression of Gq protein. In addition, coexpression of Gq protein significantly decelerated the FRET recovery of the M3R construct but not of the P2Y1R construct upon the agonist removal. These results suggest that the effects of the Gq binding on the active conformation of the receptor differ depending on the type of GPCRs.