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Literature summary extracted from

  • Egeblad-Welin, L.; Sonntag, Y.; Eklund, H.; Munch-Petersen, B.
    Functional studies of active-site mutants from Drosophila melanogaster deoxyribonucleoside kinase. Investigations of the putative catalytic glutamate-arginine pair and of residues responsible for substrate specificity (2007), FEBS J., 274, 1542-1551.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.145 crystals of a C-terminally truncated (D20) recombinant Dm-dNK mutant E52D are grown using the vapour diffusion method by hanging drop geometry. The E52D mutant is crystallized with its feedback inhibitor dTTP. The backbone conformation remains unchanged, and coordination between D52 and the dTTP–Mg complex is observed Drosophila melanogaster

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.145 E52D markedly decreased kcat Drosophila melanogaster
2.7.1.145 E52H markedly decreased kcat Drosophila melanogaster
2.7.1.145 E52Q inactive mutant protein Drosophila melanogaster
2.7.1.145 Q81N mutation shows a 200fold and 100fold increase in Km. kcat is decreased 5fold and 2fold for dThd and dCyd Drosophila melanogaster
2.7.1.145 R105H markedly decreased kcat Drosophila melanogaster
2.7.1.145 R105K inactive mutant protein Drosophila melanogaster
2.7.1.145 Y70W mutant enzyme loses activity towards purines. Negative cooperativity towards dThd and dCyd is observed Drosophila melanogaster

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.145 0.0012
-
 2'-deoxythymidine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.0022
-
 5-(E)-(2-bromvinyl)-2'-deoxyuridine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.0023
-
 2'-deoxycytidine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.0037
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme E52D Drosophila melanogaster
2.7.1.145 0.0037
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme E52H Drosophila melanogaster
2.7.1.145 0.0038
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme E52D Drosophila melanogaster
2.7.1.145 0.0049
-
 5-(E)-(2-bromvinyl)-2'-deoxyuridine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster
2.7.1.145 0.0058
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme E52H Drosophila melanogaster
2.7.1.145 0.0089
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme R105H Drosophila melanogaster
2.7.1.145 0.0243
-
 1-(beta-D-arabinofuranosyl)-cytosine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.062
-
 1-(beta-D-arabinofuranosyl)-thymine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.113
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme R105H Drosophila melanogaster
2.7.1.145 0.205
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme Q81N Drosophila melanogaster
2.7.1.145 0.231
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme Q81N Drosophila melanogaster
2.7.1.145 0.246
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster
2.7.1.145 0.251
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster
2.7.1.145 0.357
-
 1-(beta-D-arabinofuranosyl)-thymine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster
2.7.1.145 1.441
-
 1-(beta-D-arabinofuranosyl)-cytosine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.145 Drosophila melanogaster
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.145 2'-deoxycytidine + ATP
-
 Drosophila melanogaster 2'-deoxycytidine 5'-phosphate + ADP
-
 ?
2.7.1.145 ATP + 1-(beta-D-arabinofuranosyl)-cytosine
-
 Drosophila melanogaster ADP + 1-(beta-D-arabinofuranosyl)-cytosine 5'-phosphate
-
 ?
2.7.1.145 ATP + 1-(beta-D-arabinofuranosyl)-thymine
-
 Drosophila melanogaster ADP + 1-(beta-D-arabinofuranosyl)-thymine 5'-phosphate
-
 ?
2.7.1.145 ATP + 2'-deoxythymidine
-
 Drosophila melanogaster ADP + 2'-deoxythymidine 5'-phosphate
-
 ?
2.7.1.145 ATP + 5-(E)-(2-bromvinyl)-2'-deoxyuridine
-
 Drosophila melanogaster ADP + 5-(E)-(2-bromvinyl)-2'-deoxyuridine 5'-phosphate
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.145 0.0003
-
 1-(beta-D-arabinofuranosyl)-thymine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.00071
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme E52D Drosophila melanogaster
2.7.1.145 0.00072
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme E52D Drosophila melanogaster
2.7.1.145 0.0012
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme E52H Drosophila melanogaster
2.7.1.145 0.0029
-
 1-(beta-D-arabinofuranosyl)-cytosine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.0059
-
 5-(E)-(2-bromvinyl)-2'-deoxyuridine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.0067
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme R105H Drosophila melanogaster
2.7.1.145 0.012
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme E52H Drosophila melanogaster
2.7.1.145 0.0142
-
 2'-deoxythymidine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.0162
-
 2'-deoxycytidine 37°C, pH 7.5, wild-type enzyme Drosophila melanogaster
2.7.1.145 0.055
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme R105H Drosophila melanogaster
2.7.1.145 0.4
-
 5-(E)-(2-bromvinyl)-2'-deoxyuridine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster
2.7.1.145 1
-
 1-(beta-D-arabinofuranosyl)-thymine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster
2.7.1.145 2.1
-
 1-(beta-D-arabinofuranosyl)-cytosine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster
2.7.1.145 2.3
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster
2.7.1.145 5.4
-
 2'-deoxythymidine 37°C, pH 7.5, mutant enzyme Q81N Drosophila melanogaster
2.7.1.145 6.6
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme Q81N Drosophila melanogaster
2.7.1.145 6.8
-
 2'-deoxycytidine 37°C, pH 7.5, mutant enzyme Y70W Drosophila melanogaster