Increase in the size of soluble brain adenylate cyclase with activation by guanosine 5'-(beta, gamma-imino)triphosphate.
Adenylate cyclase solubilized from bovine brain with Lubrol 12A9 or Triton X-100 can be resolved into two forms by gel filtration or sucrose density gradient centrifugation. The activity of one of these forms is not stimulatable by guanosine 5'-(beta, gamma-imino)triphosphate (Gpp(NH)p) and represents the "basal" adenylate cyclase activity. In Lubrol 12A9, this form has Mr = 330,000 (total, protein and detergent) and Mr (protein only) = 265,000. The other form of adenylate cyclase can be activated by Gpp(NH)p and has a smaller molecular weight: Mr (protein and detergent) = 293,000, Mr (protein only) = 199,000. Upon activation by Gpp(NH)p, the size of the Gpp(NH)p-responsive form of adenylate cyclase increases: Mr (protein and detergent) = 330,000, Mr (protein only) = 251,000. Similar values were obtained in Triton X-100. The kinetics of heat inactivation are different in the two forms of the enzyme. Both forms are activated about 2-fold by 5 mM MnCl2. Neither of the forms is associated with measurable low Km GTPase activity. On the basis of these studies, we propose that the catalytic unit (C) of adenylate cyclase and the guanine nucleotide regulatory unit (G/F) may exist in solution in the following rapid equilibrium: (formula: see text) We propose that activation by Gpp(NH)p stabilizes the C . G/F complex and this accounts for the greater mass of the enzyme measured after activation with Gpp(NH)p. The size of the enzyme which represents the basal activity is very similar to that of the C . G/F complex. We suggest, therefore, that the basal activity is the result of a stable association of the catalytic unit with the guanine nucleotide regulatory site.
D000262:Adenylyl Cyclases / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D002540:Cerebral Cortex / Q000201:enzymology*; D004789:Enzyme Activation; D006153:Guanosine Diphosphate / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology*; D046911:Macromolecular Substances; D008970:Molecular Weight
D046911:Macromolecular Substances; D006153:Guanosine Diphosphate; D006165:Guanylyl Imidodiphosphate; D006160:Guanosine Triphosphate; D000262:Adenylyl Cyclases