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Literature summary for 2.7.1.145 extracted from

  • Solaroli, N.; Bjerke, M.; Amiri, M.H.; Johansson, M.; Karlsson, A.
    Active site mutants of Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (2003), Eur. J. Biochem., 270, 2879-2884.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli as GST-fusion proteins Drosophila melanogaster

Protein Variants

Protein Variants Comment Organism
F114A site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme Drosophila melanogaster
F114Y site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme Drosophila melanogaster
I29H site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme Drosophila melanogaster
I29H/F114Y site-directed mutagenesis, active site mutant, inactive mutant Drosophila melanogaster
N28P site-directed mutagenesis, active site mutant, inactive mutant Drosophila melanogaster
N28P/I29H site-directed mutagenesis, active site mutant, inactive mutant Drosophila melanogaster
N28P/I29H/F114Y site-directed mutagenesis, active site mutant, inactive mutant Drosophila melanogaster
Q81D site-directed mutagenesis, active site mutant, inactive mutant Drosophila melanogaster
Q81N site-directed mutagenesis, active site mutant, altered substrate specificity compared to the wild-type enzyme Drosophila melanogaster

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0025
-
2'-deoxythymidine pH 7.6, 37°C, recombinant wild-type enzyme Drosophila melanogaster
0.0031
-
2'-deoxythymidine pH 7.6, 37°C, recombinant mutant F114A Drosophila melanogaster
0.0047
-
2'-deoxythymidine pH 7.6, 37°C, recombinant mutant Q81N Drosophila melanogaster
0.022
-
2'-deoxythymidine pH 7.6, 37°C, recombinant mutant F114Y Drosophila melanogaster
0.1
-
2'-deoxythymidine above, pH 7.6, 37°C, recombinant mutants I29H and Q81D Drosophila melanogaster
0.373
-
2'-deoxyadenosine pH 7.6, 37°C, recombinant wild-type enzyme Drosophila melanogaster
0.625
-
2'-deoxyadenosine pH 7.6, 37°C, recombinant mutant Q81N Drosophila melanogaster
0.985
-
2'-deoxyguanosine pH 7.6, 37°C, recombinant mutant Q81N Drosophila melanogaster
2
-
2'-deoxyguanosine pH 7.6, 37°C, recombinant wild-type enzyme Drosophila melanogaster

Organism

Organism UniProt Comment Textmining
Drosophila melanogaster
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant GST-fusion enzymes from Escherichia coli by glutathione affinity chromatography Drosophila melanogaster

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
substrate specificities of wild-type and mutant enzymes Drosophila melanogaster

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 1-beta-D-arabinofuranosylcytosine
-
Drosophila melanogaster ADP + 1-beta-D-arabinofuranosylcytosine 5'-phosphate
-
?
ATP + 2',2'-difluorodeoxycytidine
-
Drosophila melanogaster ADP + 2',2'-difluorodeoxycytidine 5'-phosphate
-
?
ATP + 2'-deoxyadenosine
-
Drosophila melanogaster ADP + 2'-deoxyadenosine 5'-phosphate
-
?
ATP + 2'-deoxycytidine
-
Drosophila melanogaster ADP + 2'-deoxycytidine 5'-phosphate
-
?
ATP + 2'-deoxyguanosine poor substrate for the wild-type enzyme Drosophila melanogaster ADP + 2'-deoxyguanosine 5'-phosphate
-
?
ATP + 2'-deoxythymidine
-
Drosophila melanogaster ADP + 2'-deoxythymidine 5'-phosphate
-
?
ATP + 2-chlorodeoxyadenosine
-
Drosophila melanogaster ADP + 2-chlorodeoxyadenosine 5'-phosphate
-
?
ATP + E-5-(2-bromovinyl)-2'-deoxyuridine
-
Drosophila melanogaster ADP + E-5-(2-bromovinyl)-2'-deoxyuridine phosphate
-
?
additional information substrate specificities of wild-type and mutant enzymes, overview, no or poor activity with ganciclovir Drosophila melanogaster ?
-
?

Synonyms

Synonyms Comment Organism
dNK
-
Drosophila melanogaster
multisubstrate deoxyribonucleoside kinase
-
Drosophila melanogaster

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Drosophila melanogaster

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
assay at Drosophila melanogaster

Cofactor

Cofactor Comment Organism Structure
ATP
-
Drosophila melanogaster