Homologous desensitization of ovarian luteinizing hormone/human chorionic gonadotropin-responsive adenylyl cyclase is dependent upon GTP.
Homologous desensitization of ovarian LH/hCG-responsive adenylyl cyclase in cell-free systems has been reported to be dependent upon either ATP or GTP. We investigated LH/hCG-dependent desensitization of adenylyl cyclase in rabbit or pig ovarian follicular membranes and included adenyl-5'-yl imidodiphosphate to prevent nucleotide triphosphate degradation. It was found that GTP supported LH/hCG-induced desensitization with an apparent Km of approximately 0.1 microM in rabbit or pig ovarian membranes. Other nucleotide triphosphates were 100-1000 times less potent than GTP in supporting desensitization. Several nonhydrolyzable GTP analogs and the GDP analog guanosine-5'-O-(2-thiodiphosphate) would not support hCG-induced desensitization of ovarian adenylyl cyclase. Instead, these guanine nucleotide analogs were all inhibitors of GTP-supported hormone-dependent desensitization. Cholera toxin had no effect on LH-dependent desensitization. These results establish that GTP is the preferred nucleotide for homologous desensitization of the LH/hCG-sensitive adenylyl cyclase and that the GTP-dependent mechanism differs from that typically associated with guanine nucleotide-binding proteins. The GTP-dependent mechanism of desensitization of the LH receptor distinguishes it from the ATP-dependent desensitization of the beta-adrenergic receptor.
Ekstrom
R C
RC
Department of Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611.
Hunzicker-Dunn
M
M
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002474:Cell-Free System; D002772:Cholera Toxin / Q000494:pharmacology; D005260:Female; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology*; D007700:Kinetics; D006080:Ovarian Follicle / Q000201:enzymology*; D011817:Rabbits; D012265:Ribonucleotides / Q000494:pharmacology; D013552:Swine
D012265:Ribonucleotides; D006160:Guanosine Triphosphate; D002772:Cholera Toxin; D000262:Adenylyl Cyclases