ADP-ribosylation of Gs by cholera toxin is potentiated by agonist activation of beta-adrenergic receptors in the absence of GTP.
Purified Gs is a substrate for ADP-ribosylation catalyzed by cholera toxin (CTx). In S49 cyc- membranes complemented with in vitro translated Gs alpha, the beta-adrenergic agonist isoproterenol enhanced the ADP-ribosylation rate. This effect was maximal if all guanyl nucleotides were suppressed but was blocked by the beta-adrenergic antagonist alprenolol. Enhancement was partially diminished if addition of GDP followed that of isoproterenol. When added in the absence of agonist, the GTP analogues guanosine 5'-O-(gamma-thiotriphosphate) and guanosine 5'-(beta, gamma-imido)triphosphate potentiated CTx-catalyzed ADP-ribosylation of Gs alpha consistent with their activating ADP-ribosylation factors. However, this effect was lessened when the same nucleotides were tested in the presence of agonist. Taken altogether, these results indicate that like Gt and Gi, Gs is an optimal substrate for CTx when coupled to an agonist-activated receptor and depleted of nucleotide. Therefore, coupling to the receptor and subsequent departure of the GDP turn out to be the common features underlying the sensitivity of all GTP-binding proteins to CTx-catalyzed ADP-ribosylation.
Bornancin
F
F
Centre National de la Recherche Scientifique, Institut de Pharmacologie Moléculaire et Cellulaire, Valbonne, France.
Audigier
Y
Y
Chabre
M
M
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D000526:Alprenolol / Q000494:pharmacology; D002384:Catalysis; D002460:Cell Line; D002772:Cholera Toxin / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D011943:Receptors, Adrenergic, beta / Q000187:drug effects / Q000378:metabolism*
D000318:Adrenergic beta-Agonists; D011943:Receptors, Adrenergic, beta; D000246:Adenosine Diphosphate Ribose; D006160:Guanosine Triphosphate; D000526:Alprenolol; D002772:Cholera Toxin; D019204:GTP-Binding Proteins; D007545:Isoproterenol