BRENDA - Enzyme Database show
show all sequences of 2.7.1.145

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

Egeblad-Welin, L.; Sonntag, Y.; Eklund, H.; Munch-Petersen, B.; FEBS J. 274, 1542-1551 (2007)

Data extracted from this reference:

Crystallization (Commentary)
Crystallization
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
Engineering
Amino acid exchange
Commentary
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
Commentary
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 ezyme 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
Primary Accession No. (UniProt)
Commentary
Textmining
Drosophila melanogaster
-
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2'-deoxycytidine + ATP
-
673584
Drosophila melanogaster
2'-deoxycytidine 5'-phosphate + ADP
-
-
-
?
ATP + 1-(beta-D-arabinofuranosyl)-cytosine
-
673584
Drosophila melanogaster
ADP + 1-(beta-D-arabinofuranosyl)-cytosine 5'-phosphate
-
-
-
?
ATP + 1-(beta-D-arabinofuranosyl)-thymine
-
673584
Drosophila melanogaster
ADP + 1-(beta-D-arabinofuranosyl)-thymine 5'-phosphate
-
-
-
?
ATP + 2'-deoxythymidine
-
673584
Drosophila melanogaster
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
?
ATP + 5-(E)-(2-bromvinyl)-2'-deoxyuridine
-
673584
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
Commentary
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 ezyme 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
Crystallization (Commentary) (protein specific)
Crystallization
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
Engineering (protein specific)
Amino acid exchange
Commentary
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] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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 ezyme 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
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2'-deoxycytidine + ATP
-
673584
Drosophila melanogaster
2'-deoxycytidine 5'-phosphate + ADP
-
-
-
?
ATP + 1-(beta-D-arabinofuranosyl)-cytosine
-
673584
Drosophila melanogaster
ADP + 1-(beta-D-arabinofuranosyl)-cytosine 5'-phosphate
-
-
-
?
ATP + 1-(beta-D-arabinofuranosyl)-thymine
-
673584
Drosophila melanogaster
ADP + 1-(beta-D-arabinofuranosyl)-thymine 5'-phosphate
-
-
-
?
ATP + 2'-deoxythymidine
-
673584
Drosophila melanogaster
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
?
ATP + 5-(E)-(2-bromvinyl)-2'-deoxyuridine
-
673584
Drosophila melanogaster
ADP + 5-(E)-(2-bromvinyl)-2'-deoxyuridine 5'-phosphate
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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 ezyme 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
Other publictions for EC 2.7.1.145
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
738739
Kamiya
In vivo selection of active de ...
Drosophila melanogaster
J. Biotechnol.
228
52-57
2016
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1
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1
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1
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1
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3
-
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1
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-
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3
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1
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1
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1
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1
1
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1
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1
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1
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1
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3
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1
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1
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1
1
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-
-
738781
Slot Christiansen
Non-viral deoxyribonucleoside ...
Anopheles gambiae, Bombyx mori, Drosophila melanogaster
J. Genet. Genomics
42
235-248
2015
-
2
-
1
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1
3
-
18
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3
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33
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3
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2
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3
1
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1
3
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18
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33
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7
7
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739211
Jiang
Tanshinone IIA enhances bystan ...
Drosophila melanogaster
Oncol. Rep.
34
1487-1493
2015
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1
1
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4
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1
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3
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1
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4
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1
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1
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3
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1
1
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737404
Serra
-
Immobilized Drosophila melanog ...
Drosophila melanogaster
Adv. Synth. Catal.
356
563-570
2014
-
1
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1
-
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1
-
1
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1
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14
-
1
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1
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1
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1
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1
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1
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1
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1
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14
-
1
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1
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703396
Betham
Use of Drosophila deoxynucleos ...
Drosophila melanogaster
DNA Repair
9
153-160
2010
-
-
1
-
-
-
-
-
-
-
-
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-
4
-
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1
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1
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1
1
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1
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1
1
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721775
Ito
Cytotoxic effect of Drosophila ...
Drosophila melanogaster
Biol. Pharm. Bull.
33
1223-1227
2010
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1
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5
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702047
Knecht
Drosophila melanogaster deoxyr ...
Drosophila melanogaster
Biochem. Biophys. Res. Commun.
382
430-433
2009
-
-
-
1
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1
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3
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1
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1
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1
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1
1
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1
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1
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1
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-
1
1
692289
Mikkelsen
Structural studies of nucleosi ...
Drosophila melanogaster
FEBS J.
275
2151-2160
2008
-
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1
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2
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1
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694476
Clausen
A multisubstrate deoxyribonucl ...
Arabidopsis thaliana
Nucleic Acids Symp. Ser.
52
489-490
2008
-
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1
-
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2
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7
-
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1
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7
-
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-
-
-
-
-
673584
Egeblad-Welin
Functional studies of active-s ...
Drosophila melanogaster
FEBS J.
274
1542-1551
2007
-
-
-
1
7
-
-
18
-
-
-
-
-
2
-
-
-
-
-
-
-
-
5
-
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18
-
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1
7
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18
-
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5
-
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-
18
-
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-
673848
Knecht
Drosophila deoxyribonucleoside ...
Drosophila melanogaster
Gene Ther.
14
1278-1286
2007
-
1
1
-
2
-
-
-
-
-
-
-
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4
-
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1
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1
1
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672971
Legent
In vivo analysis of Drosophila ...
Drosophila melanogaster
Cell Cycle
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676247
Kamiya
Transient expression of Drosop ...
Drosophila melanogaster
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Welin
Structural basis for the chang ...
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Piskur
Animal deoxyribonucleoside kin ...
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662990
Solaroli
Investigation of the substrate ...
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660855
Krawiec
Inorganic tripolyphosphate (PP ...
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2003
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661072
Mikkelsen
Structural basis for feedback ...
Drosophila melanogaster
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Solaroli
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9
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9
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Knecht
Mosquito has a single multisub ...
Anopheles gambiae
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662985
Krawiec
Ability of adenosine-2'(3')-de ...
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2003
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661453
Eriksson
Structure and function of cell ...
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661583
Knecht
A few amino acid substitutions ...
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Knecht
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Zheng
Nucleoside analog cytotoxicity ...
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Johansson
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636842
Munch-Petersen
Functional expression of a mul ...
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1
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636843
Balzarini
The multifunctional deoxynucle ...
Drosophila melanogaster
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Knecht
Identification of residues inv ...
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636841
Johansson
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636839
Munch-Petersen
The single deoxynucleoside kin ...
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636840
Munch-Petersen
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