Mode of interaction between forskolin and manganese ion in activating catalytic unit of adenylate cyclase from rat brain.
Rat brain adenylate cyclase was solubilized with a combination of 0.7% sodium cholate and 0.6 M ammonium sulfate, and fractionated by addition of solid ammonium sulfate. The precipitate at 35% ammonium sulfate saturation contained neither guanine nucleotide-binding regulatory protein (G protein) nor calmodulin, and was used as the catalytic unit of the enzyme system. This catalytic unit was activated synergistically by forskolin and Mn2+. An apparent Km value for Mg-adenosine triphosphate (ATP) of the catalytic unit was about 80 microM in the basal state, while it increased in concurrence with the increase in the enzyme activity when forskolin was added to the assay system. The increase in the Km value depended on the forskolin concentration up to 1 microM, above which the value converged on ca. 200 microM. Furthermore, activation of the catalytic unit by forskolin was more marked at higher concentration of Mg-ATP. On the other hand, Mn2+ suppressed the increase in the Km value for Mg-ATP by forskolin, though the value in the basal state was not changed by Mn2+ alone. These findings indicate that the activation of the catalytic unit by forskolin is accompanied by the change in the affinity for Mg-ATP and Mn2+ modifies the change.
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002384:Catalysis; D005576:Colforsin; D003402:Creatine Kinase / Q000378:metabolism; D000242:Cyclic AMP / Q000494:pharmacology; D004224:Diterpenes / Q000494:pharmacology*; D004229:Dithiothreitol / Q000494:pharmacology; D004347:Drug Interactions; D004789:Enzyme Activation / Q000187:drug effects; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D015636:Magnesium Chloride; D008345:Manganese / Q000494:pharmacology*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010725:Phosphocreatine / Q000494:pharmacology; D051381:Rats
D004224:Diterpenes; D009419:Nerve Tissue Proteins; D010725:Phosphocreatine; D015636:Magnesium Chloride; D005576:Colforsin; D006165:Guanylyl Imidodiphosphate; D008345:Manganese; D000242:Cyclic AMP; D003402:Creatine Kinase; D000262:Adenylyl Cyclases; D008274:Magnesium; D004229:Dithiothreitol