show 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
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
Amino acid exchange
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
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.145
Drosophila melanogaster
-
-
-
Purification (Commentary)
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]
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
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]
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
Amino acid exchange
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
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