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Literature summary extracted from

  • Hazra, S.; Ort, S.; Konrad, M.; Lavie, A.
    Structural and kinetic characterization of human deoxycytidine kinase variants able to phosphorylate 5-substituted deoxycytidine and thymidine analogues (2010), Biochemistry, 49, 6784-6790.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
2.7.1.74 medicine ability of the designer enzymes to activate 5-substitued pyrimidines makes it possible to utilize such nucleoside analogs in suicide gene therapy or protein therapy applications that target cancer cells Homo sapiens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.74 expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain C41(DE3) Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.74 purified recombinanat Wild-type enzyme in complex with 5-methyldeoxycytidine and ADP, hanging drop vapour diffusion method, mixing of 0.001 ml of 8 mg/ml protein mixed with 0.001 ml reservoir solution containing 0.9-1.5 M trisodium citrate dihydrate and 100 mM Tris, pH 7.5, room temperature, X-ray diffraction structure determination and analysis at 1.96 A resolution Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.74 R104I/D133A site-directed mutagenesis, the mutations render the enzyme active with 5-substituted deoxycytidine and thymidine, altered substrate specificity compared to the wild-type enzyme Homo sapiens
2.7.1.74 R104L/D133A site-directed mutagenesis, the mutations render the enzyme active with 5-substituted deoxycytidine and thymidine, altered substrate specificity compared to the wild-type enzyme Homo sapiens
2.7.1.74 R104M/D133A site-directed mutagenesis, the mutations render the enzyme active with 5-substituted deoxycytidine and thymidine, altered substrate specificity compared to the wild-type enzyme Homo sapiens
2.7.1.74 R104Q/D133A site-directed mutagenesis, the mutations render the enzyme active with 5-substituted deoxycytidine and thymidine, altered substrate specificity compared to the wild-type enzyme Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.74 Mg2+ required Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.74 ATP + 5-methyldeoxycytidine Homo sapiens
-
 ADP + 5-methyl-dCMP
-
 ?
2.7.1.74 ATP + deoxycytidine Homo sapiens
-
 ADP + dCMP
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.74 Homo sapiens P27707
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.74 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain C41(DE3) by nickel affinity chromatography and gel filtration Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.74 ATP + (E)-5-(2-bromovinyl)-2'-deoxyuridine substrate of enzyme mutant R104M/D133A, poor substrate of the wild-type enzyme Homo sapiens ADP + ?
-
 ?
2.7.1.74 ATP + 5-bromodeoxycytidine
-
 Homo sapiens ADP + 5-bromo-dCMP
-
 ?
2.7.1.74 ATP + 5-iododeoxycytidine
-
 Homo sapiens ADP + 5-iodo-dCMP
-
 ?
2.7.1.74 ATP + 5-methyldeoxycytidine
-
 Homo sapiens ADP + 5-methyl-dCMP
-
 ?
2.7.1.74 ATP + 5-propynyldeoxycytidine
-
 Homo sapiens ADP + 5-propynyl-dCMP
-
 ?
2.7.1.74 ATP + D-deoxycytidine
-
 Homo sapiens ADP + D-dCMP
-
 ?
2.7.1.74 ATP + D-deoxythymidine
-
 Homo sapiens ADP + D-dTMP
-
 ?
2.7.1.74 ATP + D-deoxyuridine
-
 Homo sapiens ADP + D-dUMP
-
 ?
2.7.1.74 ATP + deoxycytidine
-
 Homo sapiens ADP + dCMP
-
 ?
2.7.1.74 ATP + gemcitabine
-
 Homo sapiens ADP + phospho-gemcitabine
-
 ?
2.7.1.74 ATP + L-deoxyuridine
-
 Homo sapiens ADP + L-dUMP
-
 ?
2.7.1.74 ATP + L-thymidine
-
 Homo sapiens ADP + L-dTMP
-
 ?
2.7.1.74 additional information substrate specificity or wild-type an dmutant enzymes, overview Homo sapiens ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.1.74 dCK
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.74 37
-
 assay at Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.74 7.5
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.74 ATP
-
 Homo sapiens

General Information

EC Number General Information Comment Organism
2.7.1.74 additional information structure-function analysis and substrate specificity, overview Homo sapiens