Multiple G-protein coupling of the dog thyrotropin receptor.
We investigated, in dog thyroid membranes, the ability of the dog thyrotropin (TSH) receptor to interact with the endogenous G proteins expressed in this tissue. Activation of the receptor led to increased incorporation of the photoreactive GTP analog [alpha-(32)P]GTP azidoanilide into immunoprecipitated alpha subunits of three G protein families: G(s), G(q/11), G(i/o). This effect was not due to a general loss of receptor G protein specificity since carbamylcholine, in the same membrane preparations, only stimulated the binding of the GTP analog to the alpha subunits of G(q/11) proteins. To investigate the multiple coupling of the dog TSH receptor in intact cells, cyclic AMP accumulation, IP(3) formation and (45)Ca2+ efflux experiments were performed. When thyrocytes were pretreated with pertussis toxin (PTX), the TSH receptor-mediated accumulation of cAMP increased by approximately 45% with TSH at 1 mU/ml, suggesting that the TSH receptor coupled to both G(s) and G(i) in vivo. On the other hand, no increase in IP(3) accumulation nor Ca2+ efflux was observed in the presence of thyrotropin. These data in intact cells are thus in contradiction with those obtained in membranes, suggesting that receptor-mediated transmembrane signalling may implicate a specificity which itself may reflect a localization and organization of the different components (receptors, G proteins, ...) in the plasma membrane of intact cells. As in some cells, G(i) activates mitogenesis by hormone activated G-protein-coupled receptors, we tested its role in the stimulation by TSH of the proliferation of thyrocytes. This was not affected by PTX, suggesting that the mitogenic effect of TSH does not involve G(i)-proteins.
Allgeier
A
A
Institute of Interdisciplinary Research, University of Brussels, Belgium.
Laugwitz
K L
KL
Van Sande
J
J
Schultz
G
G
Dumont
J E
JE
D000818:Animals; D002118:Calcium / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D004285:Dogs; D019204:GTP-Binding Proteins / Q000378:metabolism*; D015151:Immunoblotting; D011989:Receptors, Thyrotropin / Q000378:metabolism*; D015398:Signal Transduction; D013961:Thyroid Gland / Q000378:metabolism*; D013972:Thyrotropin / Q000378:metabolism*
D011989:Receptors, Thyrotropin; D013972:Thyrotropin; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D002118:Calcium