Mn2+-uncoupling of the catecholamine-sensitive adenylate cyclase system of rat reticulocytes. Parallel effects on cholera toxin-catalyzed ADP-ribosylation of the system.
High concentrations of Mn2+ interfere with functional interactions between the GTP-binding regulatory protein (G) and the catalytic moiety (C) of adenylate cyclase without perturbing interactions between receptor (R) and component G in rat reticulocyte membranes. The ability of cholera toxin to ADP-ribosylate component G and to enhance GTP-stimulated adenylate cyclase activity also appears to be correlated with the efficacy of the communication of component G with the adenylate cyclase system. Thus, increasing the concentration of Mn2+ in rat reticulocyte membrane during in vitro incubations causes a parallel loss of Gpp(NH)p-stimulated adenylate cyclase activity, cholera toxin-catalyzed [32P]ADP-ribosylation of the 42 000 Mr subunit of component G and cholera toxin-catalyzed enhancement of GTP-sensitive adenylate cyclase activity. Removal of Mn2+ by washing the membranes completely restores the sensitivity of adenylate cyclase activity. Removal of Mn2+ by washing the membranes completely restores the sensitivity of adenylate cyclase to all effectors, including cholera toxin. The data suggest that exposure of membranes to Mn2+ provides a useful tool for reversibly uncoupling catecholamine-sensitive adenylate cyclase systems. The data also suggest that the extent of cholera toxin-catalyzed ADP.-ribosylation of membrane substrates, i.e., the G component may rely on functional communication among the various components of the adenylate cyclase system. A corollary of the latter is that the amount of [32P]ADP-ribose-product detected in a membrane may reflect both the quantity and coupling efficiency of component G.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002395:Catecholamines / Q000494:pharmacology*; D002772:Cholera Toxin / Q000494:pharmacology*; D004082:Dihydroalprenolol / Q000378:metabolism; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D007545:Isoproterenol / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology*; D008970:Molecular Weight; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D051381:Rats; D012156:Reticulocytes / Q000187:drug effects / Q000201:enzymology*
D002395:Catecholamines; D009702:Nucleoside Diphosphate Sugars; D000246:Adenosine Diphosphate Ribose; D006165:Guanylyl Imidodiphosphate; D008345:Manganese; D004082:Dihydroalprenolol; D002772:Cholera Toxin; D000262:Adenylyl Cyclases; D008274:Magnesium; D007545:Isoproterenol