Biochemistry of terminal deoxynucleotidyltransferase. Identification and unity of ribo- and deoxyribonucleoside triphosphate binding site in terminal deoxynucleotidyltransferase
Terminal deoxynucleotidyltransferase is the only DNA polymerase that is strongly inhibited in the presence of ATP. We have labeled calf terminal deoxynucleotidyltransferase with (/sup 32/P)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 (/sup 32/P)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. 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.
- Research Organization:
- Univ. of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark (USA)
- OSTI ID:
- 6500384
- Journal Information:
- J. Biol. Chem.; (United States), Vol. 264:2
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ATP
CROSS-LINKING
DNA POLYMERASES
RECEPTORS
AMINO ACID SEQUENCE
BIOCHEMICAL REACTION KINETICS
BIOCHEMISTRY
CALVES
PHOSPHORUS 32
TRACER TECHNIQUES
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CATTLE
CHEMICAL REACTIONS
CHEMISTRY
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
ENZYMES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
MAMMALS
MEMBRANE PROTEINS
MOLECULAR STRUCTURE
NUCLEI
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
POLYMERIZATION
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RUMINANTS
TRANSFERASES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques