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Literature summary for 2.7.1.145 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
Search for more literature for 2.7.1.145

Crystallization (Commentary)

Crystallization (Comment) Organism
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

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

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Drosophila melanogaster
-
-
-

Substrates and Products (Substrate)

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

Turnover Number [1/s]

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