Differential signalling of both wild-type and Thr(343)Arg dopamine D(2short) receptor by partial agonists in a...

G-protein activation and Ca(2+) responses by the wild-type D(2short) receptor and a mutation Thr(343)Arg, in the distal BBXXB motif of its third intracellular loop, were investigated in CHO-K1 cells in terms of ligand:receptor:G-protein interactions. No evidence was obtained for constitutive, agonist-independent receptor activation, but differences in the ligand-mediated activation profiles of both the wild-type and mutant Thr(343)Arg D(2short) receptor were observed. Most of the partial agonists, but not bromocriptine, displayed an enhanced response at the mutant D(2short) receptor, suggesting that the mutation brings the receptor in a more active state. This enhancement was apparent both at the level of G-protein activation ([35S]GTPgammaS binding) and at the effector (Ca(2+) response) and occurred with different G(alpha)-proteins. Partial agonists were also found to act differently via the wild-type D(2short) receptor depending on the involved G(alpha)-protein. Compared with higher efficacy agonists, partial agonists displayed Ca(2+) responses with slower and dissimilar kinetic properties. Lisuride and in particular bromocriptine produced a more potent response in the co-presence of a G(alphao) protein instead of a chimeric G(alphaq/o)- or a promiscuous G(alpha15)-protein. S(+)-propylnorapomorphine showed a similar partial response irrespective of the combined G(alpha)-protein. Bromerguride and (+)-UH 232 induced weak (16 to 21% versus dopamine) intrinsic activity in the co-presence of a G(alphaq/o)-protein in contrast to their silent properties with a G(alpha15)- or a G(alphao)Cys(351)Ile-protein. In conclusion, the present data strongly suggest that multiple activation binding sites are involved with these ligands at the D(2short) receptor, and that their activation may be unravelled by either the mutation or co-expressed G(alpha)-proteins being investigated.