Studies of the guanylate cyclase of the social amoeba Dictyostelium discoideum.
Observations on the properties of the guanylate cyclase (GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2) of the social amoeba Dictyostelium discoideum are reported. On the basis of similarities in kinetic and fractionation properties, it is shown that the activity from vegetative cells and the sixfold higher activity from starved cells appear to be due to the same enzyme. Most of the activity is found to be soluble, and by gel exclusion chromatography a molecular weight of 250,000 has been estimated for this form. As the enzyme shows considerably more activity with Mn+2 than Mg+2, the Km for Mn+2 activation was determined (700 microM), and compared to the levels of total cell Mn+2 (10 microM) and Mg+2 (3mM). These data suggest that Mg+2 is probably the physiological cofactor. A previous report [J. M. Mato, (1979) Biochem. Biophys. Res. Commun. 88, 569-574] that the enzyme is activated about twofold by ATP was confirmed; but contrary to that report, activation by the ATP analog 5'-adenylyl-imidodiphosphate was also obtained. Since this analog does not donate its phosphate in kinase reactions, it is likely that ATP activates the guanylate cyclase by direct binding rather than by phosphorylation. The known in vivo agonist of the guanylate cyclase, cAMP, did not activate the enzyme in vitro, either alone or in various combinations with calcium, calmodulin, ATP, and phospholipids.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D000242:Cyclic AMP / Q000494:pharmacology; D004023:Dictyostelium / Q000201:enzymology* / Q000502:physiology; D004355:Drug Stability; D004789:Enzyme Activation / Q000187:drug effects; D006162:Guanylate Cyclase / Q000302:isolation & purification / Q000378:metabolism*; D008345:Manganese / Q000494:pharmacology
D008345:Manganese; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D006162:Guanylate Cyclase; D002118:Calcium