EC Number | Cloned (Comment) | Organism |
---|---|---|
2.7.4.13 | gene dnk, recombinant expression of wild-type and mutant enzymes in Escherichia coli strains BL21(DE3) and XL10Gold | Tequintavirus T5 |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.7.4.13 | D16N | site-directed mutagenesis of the P loop-related residue, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | D170N | site-directed mutagenesis of the core residue, almost inactive mutant | Tequintavirus T5 |
2.7.4.13 | E176Q | site-directed mutagenesis of the core residue, almost inactive mutant | Tequintavirus T5 |
2.7.4.13 | G137A | site-directed mutagenesis of the dNMP-binding residue, the mutant shows highly reduced activity and altered substrate specificity compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | K131E | site-directed mutagenesis, of the dNMP-binding residue, the mutant shows slightly increased activity and altered substrate specificity compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | additional information | all enzyme mutants but T17S show an increased pH optimum compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | Q134A | site-directed mutagenesis of the dNMP-binding residue, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | R130K | site-directed mutagenesis of the dNMP-binding residue, almost inactive mutant | Tequintavirus T5 |
2.7.4.13 | R172I | site-directed mutagenesis of the core residue, almost inactive mutant | Tequintavirus T5 |
2.7.4.13 | S13A | site-directed mutagenesis of the P loop-related residue, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | T138A | site-directed mutagenesis of the dNMP-binding residue, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | T17N | site-directed mutagenesis of the P loop-related residue, the mutant shows highly reduced activity and altered substrate specificity compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | T17S | site-directed mutagenesis, of the P loop-related residue, the mutant shows reduced activity compared to the wild-type enzyme | Tequintavirus T5 |
2.7.4.13 | W150A | site-directed mutagenesis of the core residue | Tequintavirus T5 |
2.7.4.13 | W150F | site-directed mutagenesis of the conserved residue, the mutant shows unaltered activity compared to the wild-type enzyme | Tequintavirus T5 |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
2.7.4.13 | Mg2+ | required | Tequintavirus T5 |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.7.4.13 | ATP + dAMP | Tequintavirus T5 | - |
ADP + dADP | - |
? | |
2.7.4.13 | ATP + dCMP | Tequintavirus T5 | - |
ADP + dCDP | - |
? | |
2.7.4.13 | ATP + dGMP | Tequintavirus T5 | - |
ADP + dGDP | - |
? | |
2.7.4.13 | ATP + dTMP | Tequintavirus T5 | - |
ADP + dTDP | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.7.4.13 | Tequintavirus T5 | Q6QGP4 | gene dnk | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.7.4.13 | recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange and affinity chromatography | Tequintavirus T5 |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
2.7.4.13 | additional information | - |
specific activties of wild-type enzyme and enzyme mutants with different dNMP acceptor substrates, overview | Tequintavirus T5 |
2.7.4.13 | 25.78 | - |
pH 7.5, 37°C, recombinant wild-type enzyme, substrate dCMP | Tequintavirus T5 |
2.7.4.13 | 36.54 | - |
pH 7.5, 37°C, recombinant wild-type enzyme, substrate dGMP | Tequintavirus T5 |
2.7.4.13 | 41.24 | - |
pH 7.5, 37°C, recombinant wild-type enzyme, substrate dTMP | Tequintavirus T5 |
2.7.4.13 | 73.17 | - |
pH 7.5, 37°C, recombinant wild-type enzyme, substrate dAMP | Tequintavirus T5 |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.7.4.13 | ATP + dAMP | - |
Tequintavirus T5 | ADP + dADP | - |
? | |
2.7.4.13 | ATP + dAMP | best substrate dAMP, wild-type enzyme | Tequintavirus T5 | ADP + dADP | - |
? | |
2.7.4.13 | ATP + dCMP | - |
Tequintavirus T5 | ADP + dCDP | - |
? | |
2.7.4.13 | ATP + dCMP | 35.2% of the activity with dAMP, wild-type enzyme | Tequintavirus T5 | ADP + dCDP | - |
? | |
2.7.4.13 | ATP + dGMP | - |
Tequintavirus T5 | ADP + dGDP | - |
? | |
2.7.4.13 | ATP + dGMP | 49.9% of the activity with dAMP, wild-type enzyme | Tequintavirus T5 | ADP + dGDP | - |
? | |
2.7.4.13 | ATP + dTMP | - |
Tequintavirus T5 | ADP + dTDP | - |
? | |
2.7.4.13 | ATP + dTMP | 56.4% of the activity with dAMP, wild-type enzyme | Tequintavirus T5 | ADP + dTDP | - |
? | |
2.7.4.13 | additional information | substrate specificities of wild-type and mutant enzymes, structure-function analysis, overview | Tequintavirus T5 | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
2.7.4.13 | monomer | - |
Tequintavirus T5 |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.7.4.13 | deoxyribonucleoside monophosphate kinase | - |
Tequintavirus T5 |
2.7.4.13 | dNMPK | - |
Tequintavirus T5 |
2.7.4.13 | T5 bacteriophage dNMPK | - |
Tequintavirus T5 |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
2.7.4.13 | 25 | - |
assay at | Tequintavirus T5 |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
2.7.4.13 | 7 | - |
wild-type enzyme and mutant T17S | Tequintavirus T5 |
2.7.4.13 | 7.5 | 8.5 | all enzyme mutants but T17S | Tequintavirus T5 |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
2.7.4.13 | ATP | - |
Tequintavirus T5 |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.7.4.13 | additional information | essential catalytic site residue, active site modelling, overview. The charged amino acid residues of the NMP binding domain and the presence of an OH-group at position 17 are important for the catalytic activity. Arginine residues at positions 130 and 172 are involved in the binding to the donor gamma-phosphoryl and acceptor alpha-phosphoryl groups, as well as the aspartic acid residue at position 16 of the ATP-binding site (P-loop), in the binding to some acceptors, first of all dTMP. The NMPK-P loop, or a glycine enriched loop, forms an anion hole interacting with donor phosphoryl groups. It contains a GX1X2X3X4GKX5T(S) consensus called the Walker A motif | Tequintavirus T5 |