Stimulatory and inhibitory regulation of myocardial adenylate cyclase by 5'-guanylyl-imidodiphosphate.
Particulate and soluble rat myocardial adenylate cyclase enzymes were characterized with respect to their stimulatory and inhibitory regulation by Gpp(NH)p. Gpp(NH)p (60 microM) stimulated Mg2+- and Mn2+-dependent adenylate cyclase. High concentrations of Gpp(NH)p (600 microM) attenuated the maximal stimulatory response to Gpp(NH)p but only at low cation concentrations. The attenuating effects of 600 microM Gpp(NH)p resulted predominantly from the introduction of a prolonged lag in the kinetics of activation of adenylate cyclase. Steady-state rates of adenylate cyclase activities were similar with either 60 or 600 microM Gpp(NH)p. At any concentration of Gpp(NH)p, the lag was eliminated by Mg ions or isoproterenol. No antihysteretic property for free Mn ions was evident. Forskolin-sensitive particulate adenylate cyclase was not stimulated further by Gpp(NH)p. A 600 microM concentration of Gpp(NH)p inhibited particulate forskolin-sensitive adenylate cyclase at low Mg ion concentrations. In contrast, Gpp(NH)p at 60 microM consistently activated forskolin-sensitive adenylate cyclase after solubilization. The early transient inhibitory properties of 600 microM Gpp(NH)p which resulted in attenuation of adenylate cyclase by 600 microM Gpp(NH)p were diminished by detergent extraction, resulting in only a minor effect of 600 microM Gpp(NH)p to inhibit solubilized adenylate cyclase. These findings indicate that guanine nucleotides exert both stimulatory and inhibitory control upon the myocardial adenylate cyclase enzyme; that solubilization shifts the balance between the stimulatory and inhibitory properties of Gpp(NH)p to allow more dominant expression of the stimulatory response; and that Mg ions critically modify the nature of the myocardial adenylate cyclase response to Gpp(NH)p.
D000262:Adenylyl Cyclases / Q000032:analysis*; D000818:Animals; D005576:Colforsin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000032:analysis; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives*; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology*; D066298:In Vitro Techniques; D007545:Isoproterenol / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D009206:Myocardium / Q000201:enzymology*; D051381:Rats
D005576:Colforsin; D006165:Guanylyl Imidodiphosphate; D008345:Manganese; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D008274:Magnesium; D007545:Isoproterenol