On the interaction of 3,4,5,6-tetrahydrouridine with human liver cytidine deaminase.
In contrast to the rapid inhibition of bacterial cytidine deaminase by 3,4,5,6-tetrahydrouridine, the onset of inhibition of the enzyme from human liver was found to be relatively slow. Inhibition was found to be reversible, and the corrected rate constants for binding (kon = 2.4 x 10(4) M-1 sec-1) and release (koff = 5.6 x 10(-4) sec-1) were in reasonable agreement with a Ki value (2.9 x 10(-8) M) measured separately under steady-state conditions, which was several orders of magnitude lower than estimates previously reported in the literature. Rates of binding and release of this potential transition state analogue were not appreciably affected by the substitution of deuterium oxide for solvent water. The slow onset of inhibition, which was also observed for cytidine deaminase from HeLa cells, suggests that structural reorganization precedes the formation of a stable enzyme-inhibitor complex. 6-Azacytidine, which favors a "high-anti" configuration at the glycosidic bond, was found to be active as a substrate for cytidine deaminase, with a turnover number exceeding that of cytidine. 2,2'-Anhydro-1-beta-D-arabinofuranosylcytosine, which is restricted to the "syn" configuration, was found to be without activity as a substrate or an inhibitor.
D001374:Azacitidine / Q000494:pharmacology; D001665:Binding Sites; D003561:Cytarabine / Q000031:analogs & derivatives / Q000494:pharmacology; D003564:Cytidine Deaminase / Q000037:antagonists & inhibitors*; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D009700:Nucleoside Deaminases / Q000037:antagonists & inhibitors*; D011485:Protein Binding; D013767:Tetrahydrouridine / Q000494:pharmacology*; D014529:Uridine / Q000031:analogs & derivatives*
D003561:Cytarabine; D013767:Tetrahydrouridine; D009700:Nucleoside Deaminases; D003564:Cytidine Deaminase; D001374:Azacitidine; D014529:Uridine