title: Biochemistry of terminal deoxynucleotidyltransferase. Identification and unity of ribo- and deoxyribonucleoside triphosphate binding site in terminal deoxynucleotidyltransferase.
abstract: Terminal deoxynucleotidyltransferase is the only DNA polymerase that is strongly inhibited in the presence of ATP. We have labeled calf terminal deoxynucleotidyltransferase with [32P]ATP in order to identify its binding site in terminal deoxynucleotidyltransferase. The specificity of ATP cross-linking to terminal deoxynucleotidyltransferase is shown by the competitive inhibition of the overall cross-linking reaction by deoxynucleoside triphosphates, as well as the ATP analogs Ap4A and Ap5A. Tryptic peptide mapping of [32P]ATP-labeled enzyme revealed a peptide fraction that contained the majority of cross-linked ATP. The properties, chromatographic characteristics, amino acid composition, and sequence analysis of this peptide fraction were identical with those found associated with dTTP cross-linked terminal deoxynucleotidyl-transferase peptide (Pandey, V. N., and Modak, M. J. (1988a). J. Biol. Chem. 263, 3744-3751). The involvement of the same 2 cysteine residues in the crosslinking of both nucleotides further confirmed the unity of the ATP and dTTP binding domain that contains residues 224-237 in the primary amino acid sequence of calf terminal deoxynucleotidyltransferase.
authors.affiliation: Department of Biochemistry, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark 07103.
authors.forename: V N
authors.initials: VN
authors.lastname: Pandey
authors.forename: M J
authors.initials: MJ
authors.lastname: Modak
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism* / Q000528:radiation effects; D003854:Deoxyribonucleotides / Q000494:pharmacology*; D007700:Kinetics; D010446:Peptide Fragments / Q000032:analysis; D012265:Ribonucleotides / Q000494:pharmacology*; D014466:Ultraviolet Rays
chemical_list: D000596:Amino Acids; D003854:Deoxyribonucleotides; D010446:Peptide Fragments; D012265:Ribonucleotides; D000255:Adenosine Triphosphate; D004253:DNA Nucleotidylexotransferase