Characterization of the purine-reactive site of the rat testis cytosolic adenylate cyclase.
Naturally soluble rat germ cell adenylate cyclase was inhibited by adenosine and the adenosine analogs, 9-beta-D-arabinofuranosyl adenine (AFA) and 2',5'-dideoxyadenosine (DDA), all of which inhibited hormone-sensitive adenylate cyclases at the "P" site. The IC50 values for adenosine and DDA were approximately 0.1 and for AFA, 4.0 mM. The onset of adenosine inhibition was very rapid whether adenosine was added to the enzyme reactant mixture at time zero concomitantly with the addition of substrate or after the enzyme had been activated by the addition of substrate. The adenosine analogs, N6-methyladenosine (MeA) and N6-phenylisopropyl adenosine (PIA), which interact with plasma membrane receptors ("R" receptors) for hormone-sensitive adenylate cyclase, had little effect on the activity of the cytosolic adenylate cyclase. Additionally, aminophylline, which has been shown to competitively antagonize adenosine interactions with the plasma membrane "R" receptors but not "P" site interactions, had no effect upon substrate activation of the soluble enzyme and did not prevent adenosine from inhibiting the activity of the enzyme. These data provide evidence for an adenosine regulatory site on the cytosolic enzyme which resembles the "P" site described for membrane bound-adenylate cyclase.
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D003600:Cytosol / Q000201:enzymology; D003839:Deoxyadenosines / Q000031:analogs & derivatives / Q000494:pharmacology; D016048:Dideoxyadenosine / Q000031:analogs & derivatives*; D007700:Kinetics; D008297:Male; D010660:Phenylisopropyladenosine / Q000494:pharmacology; D011687:Purines / Q000378:metabolism*; D051381:Rats; D013737:Testis / Q000201:enzymology*; D014740:Vidarabine / Q000494:pharmacology
D000067956:Adenylyl Cyclase Inhibitors; D003839:Deoxyadenosines; D011687:Purines; D010660:Phenylisopropyladenosine; D016048:Dideoxyadenosine; C021008:2',5'-dideoxyadenosine; C010223:N-methyladenosine; D000262:Adenylyl Cyclases; D014740:Vidarabine; D000241:Adenosine