Doxynucleotide-polymerizing enzymes of calf thymus gland. IV. Inhibition of terminal deoxynucleotidyl transferase by metal ligands.
The polymerization of deoxynucleoside triphosphates, catalyzed by terminal deoxynucleotidyl transferase from calf thymus gland, is strongly inhibited by various metal chelators. The reaction appears to be first order with respect to enzyme, deoxynucleoside triphosphate, and initiator, indicating that there is only one catalytic center for each enzyme molecule. The presence of metal chelator does not affect the molecular state of the enzyme or the order of reaction. An analysis of the kinetics of o-phenanthroline inhibition demonstrates that the ligand is noncompetitive with nucleoside triphosphate substrate and strictly competitive with oligodeoxynucleotide initiator. These results, obtained in the presence of millimolar Mg(++), suggest that the initiator binds to the enzyme through a transition metal atom and ligand inhibition occurs at that site. The precise spatial arrangement of the coordination orbitals around a metal atom provides a model to visualize the specificity of the enzyme for the 3'-OH end of the growing polynucleotide chain.
D000818:Animals; D002417:Cattle; D002614:Chelating Agents; D003545:Cysteine; D004254:DNA Nucleotidyltransferases; D004492:Edetic Acid; D005029:Ethylenediamines; D007700:Kinetics; D008274:Magnesium; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors*; D010618:Phenanthrolines; D011119:Polynucleotides; D011725:Pyridines; D013950:Thymus Gland / Q000201:enzymology; D014316:Tritium
D002614:Chelating Agents; D005029:Ethylenediamines; D010618:Phenanthrolines; D011119:Polynucleotides; D011725:Pyridines; D014316:Tritium; D004492:Edetic Acid; D004254:DNA Nucleotidyltransferases; D009713:Nucleotidyltransferases; D008274:Magnesium; D003545:Cysteine