The thermal lability of adenylate cyclase: mechanisms of stabilization.
The thermal inactivation of adenylate cyclase was investigated in human lymphocytes and in the N-protein deficient cyc- S49 mouse lymphoma cell line. The enzyme is rapidly inactivated at 37C with a t1/2 of 5.5 and 4.5 min respectively in human and cyc- membranes. Thermal inactivation is prevented by at least two mechanisms. The first mechanism involves ATP which stabilizes adenylate cyclase in a concentration dependent manner similar to the Km of ATP for cAMP formation. However, the inhibition of inactivation does not require Mg++ while the enzyme catalysis of ATP to cAMP does. The second mechanism involves substances which activate the enzyme. The human lymphocyte enzyme is equally stabilized by either NaF, GppNHp, or forskolin. In contrast, the cyc- enzyme is fully stabilized by forskolin but only partially stabilized by NaF. When human erythrocyte N-protein extract is added to cyc- membranes, NaF fully stabilizes the enzyme. These data suggest that an activated N-protein is instrumental in stabilizing adenylate cyclase and that there is some N-protein component in cyc- membranes through which NaF may be exerting its stabilizing action.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002316:Cardiotonic Agents / Q000494:pharmacology; D002460:Cell Line; D005576:Colforsin; D004224:Diterpenes / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004910:Erythrocyte Membrane / Q000201:enzymology*; D004912:Erythrocytes / Q000201:enzymology*; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology*; D008223:Lymphoma / Q000201:enzymology; D051379:Mice; D009374:Neoplasms, Experimental / Q000201:enzymology; D012969:Sodium Fluoride / Q000494:pharmacology; D013696:Temperature
D000067956:Adenylyl Cyclase Inhibitors; D002316:Cardiotonic Agents; D004224:Diterpenes; D005576:Colforsin; D006165:Guanylyl Imidodiphosphate; D000255:Adenosine Triphosphate; D012969:Sodium Fluoride; D000262:Adenylyl Cyclases