Herpes simplex virus type 1 ribonucleotide reductase: selective and synergistic inactivation by A1110U and its iron complex.
2-Acetylpyridine-5-[(dimethylamino)thiocarbonyl]thiocarbonohydr azone (A1110U) inactivated herpes simplex virus Type 1 ribonucleotide reductase (EC 1.17.4.1) by a first-order process (kinact) which had a maximum value (Mkinact) of 8 hr-1 and a Kd that was less than 1 microM. The stable complex between iron and A1110U, (A1110U)2Fe+i, inactivated this enzyme with a Mkinact of 7 hr-1 and a Kd of 7 microM. Free A1110U and its iron-complex synergized as inactivators of the enzyme. For example, the kinact for the combination of 2 microM A1110U and 1 microM (A1110U)2Fe+i as independent inactivators was calculated to be about 9 hr-1, while the observed value was 32 hr-1. The bimolecular rate constant for inactivation of the viral enzyme by (A1110U)2Fe+i in the presence of a saturating concentration of A1110U was 2.5 10(7) M-1 hr-1 at 30 degrees. Human ribonucleotide reductase was less sensitive to the inhibitory effects of A1110U and its iron-complex. This enzyme was neither inhibited nor inactivated by A1110U and was weakly inhibited by (A1110U)2Fe+i. Furthermore, inactivation required the combination of A1110U and (A1110U)2Fe+i. The bimolecular rate constant for inactivation of human ribonucleotide reductase by (A1110U)2Fe+i in the presence of a saturating concentration of A1110U was considerably smaller (3.8 10(6) M-1 hr-1 at 37 degrees) than the analogous constant for the viral enzyme. Several iron-chelating reagents with unrelated structures substituted for free A1110U in its various roles with both enzymes. However, the iron complexes of these alternative chelators did not substitute for (A1110U)2Fe+i. The rates of inactivation of both enzymes were independent of the oxidation state of iron in (A1110U)2Fe+i and of CDP concentration. The inactivated enzymes were reactivated rapidly by FeSO4, but were not reactivated by CoCl2, CuSO4, or NiCl2. MnCl2 inhibited reactivation of the viral enzyme by FeSO4.
Porter
D J
DJ
Experimental Therapy Division, Wellcome Research Laboratories, Research Triangle Park, NC 27709.
Harrington
J A
JA
Spector
T
T
D000255:Adenosine Triphosphate / Q000494:pharmacology; D003565:Cytidine Diphosphate / Q000378:metabolism / Q000494:pharmacology; D003676:Deferoxamine / Q000494:pharmacology; D004357:Drug Synergism; D004789:Enzyme Activation / Q000187:drug effects; D004793:Enzyme Reactivators; D005296:Ferrous Compounds / Q000494:pharmacology; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology*; D007501:Iron / Q000494:pharmacology*; D007502:Iron Chelating Agents / Q000494:pharmacology; D007700:Kinetics; D015636:Magnesium Chloride / Q000494:pharmacology; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010618:Phenanthrolines / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology*; D012264:Ribonucleotide Reductases / Q000037:antagonists & inhibitors*; D018139:Simplexvirus / Q000201:enzymology*
D004793:Enzyme Reactivators; D005296:Ferrous Compounds; D006835:Hydrazones; D007502:Iron Chelating Agents; D010618:Phenanthrolines; D011725:Pyridines; D015636:Magnesium Chloride; C020748:ferrous sulfate; D003565:Cytidine Diphosphate; D000255:Adenosine Triphosphate; C059415:A 1110U; D007501:Iron; D012264:Ribonucleotide Reductases; D003676:Deferoxamine; C025205:1,10-phenanthroline