Role of the carbohydrate moiety of human choriogonadotropin in its thyrotropic activity.

To investigate the role of the carbohydrate moiety of human choriogonadotropin (hCG) in its thyrotropic activity, highly purified hCG and its desialylated subunits were treated with anhydrous HF/anisole (1 h, 0 degree C). The deglycosylated alpha- and beta-subunits were recombined with their native complementary subunits, and the interactions of these hCG congeners with the thyrotropin (TSH) receptor-adenylate cyclase system were investigated using human thyroid membranes. Deglycosylated hCG (dghCG) bound to the high affinity-low capacity TSH-binding sites of thyroid membranes; its equilibrium dissociation constant was lower than that of asialo-hCG (ashCG) (ED50: 2.6 and 6 microM, respectively). Like ashCG, dghCG did not stimulate thyroidal adenylate cyclase, but rather inhibited TSH stimulation of this enzyme in a dose-dependent manner. Thus, dghCG behaved as an antagonist and exhibited an inhibition constant of 0.78 microM while ashCG exhibited a constant of 1.50 microM. As might be predicted from the behavior of dghCG, absence of carbohydrate from either subunit enhanced the ability of the hCG hybrid recombinants to interact with the TSH receptor-adenylate cyclase system. However, only the hybrid recombinant lacking carbohydrate on its alpha-subunit lacked intrinsic thyrotropic activity; the hybrid recombinant lacking carbohydrate on its beta-subunit not only displayed intrinsic thyrotropic activity, but was of even higher potency than intact hCG in stimulating thyroidal adenylate cyclase. These results demonstrate that the carbohydrate moieties of both hCG subunits impede the process of recognition of hCG by the TSH receptor, while the carbohydrate moiety of the alpha-subunit, but not that of the beta-subunit, is essential for the process of hCG activation of thyroidal adenylate cyclase.