Active site mutants of Drosophila melanogaster multisubstrate deoxyribonucleoside kinase
Solaroli, N.; Bjerke, M.; Amiri, M.H.; Johansson, M.; Karlsson, A.; Eur. J. Biochem. 270, 2879-2884 (2003) 
Data extracted from this reference:
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
2.7.1.145
expression of wild-type and mutant enzymes in Escherichia coli as GST-fusion proteins
Drosophila melanogaster
Engineering
EC Number
Protein Variants
Commentary
Organism
2.7.1.145
F114A
site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme
Drosophila melanogaster
2.7.1.145
F114Y
site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme
Drosophila melanogaster
2.7.1.145
I29H
site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme
Drosophila melanogaster
2.7.1.145
I29H/F114Y
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
N28P
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
N28P/I29H
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
N28P/I29H/F114Y
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
Q81D
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
Q81N
site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme
Drosophila melanogaster
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.145
0.0025
-
2'-deoxythymidine
pH 7.6, 37°C, recombinant wild-type enzyme
Drosophila melanogaster
2.7.1.145
0.0031
-
2'-deoxythymidine
pH 7.6, 37°C, recombinant mutant F114A
Drosophila melanogaster
2.7.1.145
0.0047
-
2'-deoxythymidine
pH 7.6, 37°C, recombinant mutant Q81N
Drosophila melanogaster
2.7.1.145
0.022
-
2'-deoxythymidine
pH 7.6, 37°C, recombinant mutant F114Y
Drosophila melanogaster
2.7.1.145
0.1
-
2'-deoxythymidine
above, pH 7.6, 37°C, recombinant mutants I29H and Q81D
Drosophila melanogaster
2.7.1.145
0.373
-
2'-deoxyadenosine
pH 7.6, 37°C, recombinant wild-type enzyme
Drosophila melanogaster
2.7.1.145
0.625
-
2'-deoxyadenosine
pH 7.6, 37°C, recombinant mutant Q81N
Drosophila melanogaster
2.7.1.145
0.985
-
2'-deoxyguanosine
pH 7.6, 37°C, recombinant mutant Q81N
Drosophila melanogaster
2.7.1.145
2
-
2'-deoxyguanosine
pH 7.6, 37°C, recombinant wild-type enzyme
Drosophila melanogaster
Organism
EC Number
Organism
UniProt
Commentary
Textmining
2.7.1.145
Drosophila melanogaster
-
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
2.7.1.145
recombinant wild-type and mutant GST-fusion enzymes from Escherichia coli by glutathione affinity chromatography
Drosophila melanogaster
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.1.145
additional information
-
substrate specificities of wild-type and mutant enzymes
Drosophila melanogaster
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2.7.1.145
ATP + 1-beta-D-arabinofuranosylcytosine
-
661629
Drosophila melanogaster
ADP + 1-beta-D-arabinofuranosylcytosine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2',2'-difluorodeoxycytidine
-
661629
Drosophila melanogaster
ADP + 2',2'-difluorodeoxycytidine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxyadenosine
-
661629
Drosophila melanogaster
ADP + 2'-deoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxycytidine
-
661629
Drosophila melanogaster
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxyguanosine
poor substrate for the wild-type enzyme
661629
Drosophila melanogaster
ADP + 2'-deoxyguanosine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxythymidine
-
661629
Drosophila melanogaster
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2-chlorodeoxyadenosine
-
661629
Drosophila melanogaster
ADP + 2-chlorodeoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + E-5-(2-bromovinyl)-2'-deoxyuridine
-
661629
Drosophila melanogaster
ADP + E-5-(2-bromovinyl)-2'-deoxyuridine phosphate
-
-
-
?
2.7.1.145
additional information
substrate specificities of wild-type and mutant enzymes, overview, no or poor activity with ganciclovir
661629
Drosophila melanogaster
?
-
-
-
-
Synonyms
EC Number
Synonyms
Commentary
Organism
2.7.1.145
dNK
-
Drosophila melanogaster
2.7.1.145
multisubstrate deoxyribonucleoside kinase
-
Drosophila melanogaster
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.145
37
-
assay at
Drosophila melanogaster
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.145
7.6
-
assay at
Drosophila melanogaster
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.145
ATP
-
Drosophila melanogaster
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.145
expression of wild-type and mutant enzymes in Escherichia coli as GST-fusion proteins
Drosophila melanogaster
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.145
ATP
-
Drosophila melanogaster
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
2.7.1.145
F114A
site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme
Drosophila melanogaster
2.7.1.145
F114Y
site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme
Drosophila melanogaster
2.7.1.145
I29H
site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme
Drosophila melanogaster
2.7.1.145
I29H/F114Y
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
N28P
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
N28P/I29H
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
N28P/I29H/F114Y
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
Q81D
site-directed mutagenesis, active site mutant, inactive mutant
Drosophila melanogaster
2.7.1.145
Q81N
site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme
Drosophila melanogaster
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.145
0.0025
-
2'-deoxythymidine
pH 7.6, 37°C, recombinant wild-type enzyme
Drosophila melanogaster
2.7.1.145
0.0031
-
2'-deoxythymidine
pH 7.6, 37°C, recombinant mutant F114A
Drosophila melanogaster
2.7.1.145
0.0047
-
2'-deoxythymidine
pH 7.6, 37°C, recombinant mutant Q81N
Drosophila melanogaster
2.7.1.145
0.022
-
2'-deoxythymidine
pH 7.6, 37°C, recombinant mutant F114Y
Drosophila melanogaster
2.7.1.145
0.1
-
2'-deoxythymidine
above, pH 7.6, 37°C, recombinant mutants I29H and Q81D
Drosophila melanogaster
2.7.1.145
0.373
-
2'-deoxyadenosine
pH 7.6, 37°C, recombinant wild-type enzyme
Drosophila melanogaster
2.7.1.145
0.625
-
2'-deoxyadenosine
pH 7.6, 37°C, recombinant mutant Q81N
Drosophila melanogaster
2.7.1.145
0.985
-
2'-deoxyguanosine
pH 7.6, 37°C, recombinant mutant Q81N
Drosophila melanogaster
2.7.1.145
2
-
2'-deoxyguanosine
pH 7.6, 37°C, recombinant wild-type enzyme
Drosophila melanogaster
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.145
recombinant wild-type and mutant GST-fusion enzymes from Escherichia coli by glutathione affinity chromatography
Drosophila melanogaster
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.1.145
additional information
-
substrate specificities of wild-type and mutant enzymes
Drosophila melanogaster
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2.7.1.145
ATP + 1-beta-D-arabinofuranosylcytosine
-
661629
Drosophila melanogaster
ADP + 1-beta-D-arabinofuranosylcytosine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2',2'-difluorodeoxycytidine
-
661629
Drosophila melanogaster
ADP + 2',2'-difluorodeoxycytidine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxyadenosine
-
661629
Drosophila melanogaster
ADP + 2'-deoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxycytidine
-
661629
Drosophila melanogaster
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxyguanosine
poor substrate for the wild-type enzyme
661629
Drosophila melanogaster
ADP + 2'-deoxyguanosine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2'-deoxythymidine
-
661629
Drosophila melanogaster
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + 2-chlorodeoxyadenosine
-
661629
Drosophila melanogaster
ADP + 2-chlorodeoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.145
ATP + E-5-(2-bromovinyl)-2'-deoxyuridine
-
661629
Drosophila melanogaster
ADP + E-5-(2-bromovinyl)-2'-deoxyuridine phosphate
-
-
-
?
2.7.1.145
additional information
substrate specificities of wild-type and mutant enzymes, overview, no or poor activity with ganciclovir
661629
Drosophila melanogaster
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.145
37
-
assay at
Drosophila melanogaster
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.145
7.6
-
assay at
Drosophila melanogaster