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Literature summary for 2.7.4.1 extracted from
- Amino acids involved in polyphosphate synthesis and its mobilization are distinct in polyphosphate kinase-1 from Mycobacterium tuberculosis (2011), PLoS ONE, 6, e27398.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| His-tagged enzyme is expressed in Escherichia coli BL21(DE3) cells | Mycobacterium tuberculosis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A615S | the mutant exhibits about wild type activity for ATP synthesis and polyphosphate synthesis | Mycobacterium tuberculosis |
| E681A | the mutant exhibits less than 5% activity for ATP synthesis and less than 30% activity for polyphosphate synthesis compared o the wild type enzyme | Mycobacterium tuberculosis |
| H150A | the mutation causes PPK1 protein deficient but not defective in autophosphorylation and severely affects ATP or polyphosphate synthesis | Mycobacterium tuberculosis |
| H150Q | the mutation causes PPK1 protein deficient but not defective in autophosphorylation and mildly affects ATP or polyphosphate synthesis | Mycobacterium tuberculosis |
| H480A | the mutant catalytic efficiency for the polyphosphate synthesis is almost or very close to that of the wild type enzyme | Mycobacterium tuberculosis |
| H480Q | the mutant catalytic efficiency for the polyphosphate synthesis is almost or very close to that of the wild type enzyme | Mycobacterium tuberculosis |
| H491A | inactive | Mycobacterium tuberculosis |
| H491Q | inactive | Mycobacterium tuberculosis |
| H510Q | the mutant catalytic efficiency for the polyphosphate synthesis is almost or very close to that of the wild type enzyme | Mycobacterium tuberculosis |
| N515A | the mutant exhibits less than 2% activity for ATP synthesis and about 65% activity for polyphosphate synthesis compared to the wild type enzyme | Mycobacterium tuberculosis |
| R431A | the mutant exhibits less than 5% activity for ATP synthesis and about 78% activity for polyphosphate synthesis compared to the wild type enzyme | Mycobacterium tuberculosis |
| R461A | the mutant exhibits less than 2% activity for ATP synthesis and about 76% activity for polyphosphate synthesis compared to the wild type enzyme | Mycobacterium tuberculosis |
| R624A | the mutant exhibits less than 2% activity for ATP synthesis and about 65% activity for polyphosphate synthesis compared to the wild type enzyme | Mycobacterium tuberculosis |
| R654A | the mutant exhibits less than 1% activity for ATP synthesis and about 90% activity for polyphosphate synthesis compared to the wild type enzyme | Mycobacterium tuberculosis |
| S668A | the mutant exhibits less than 5% activity for ATP synthesis and less than 30% activity for polyphosphate synthesis compared to the wild type enzyme | Mycobacterium tuberculosis |
| Y524A | the mutant exhibits less than 10% activity for ATP synthesis and less than 20% activity for polyphosphate synthesis compared to the wild type enzyme | Mycobacterium tuberculosis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.16 | - |
ADP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis |  |
| 0.176 | - |
ADP | mutant H510Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 0.18 | - |
ADP | mutant H480Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 0.2 | - |
ADP | mutant H480A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 1.35 | - |
ATP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 1.38 | - |
ATP | mutant H480Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 1.47 | - |
ATP | mutant H480A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 1.5 | 2 | ATP | mutant H510Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 2 | - |
ADP | mutant Y524A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 5.2 | - |
ATP | mutant H510A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 6.8 | - |
ADP | mutant H510A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 8 | - |
ADP | isoform PPK2, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 11.3 | - |
GDP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 13 | - |
ATP | mutant Y524A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | necessary for activity | Mycobacterium tuberculosis | |
| Mn2+ | necessary for activity | Mycobacterium tuberculosis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 87000 | - |
2 * 87000, isoform PPK1, SDS-PAGE | Mycobacterium tuberculosis |
| 197000 | - |
isoform PPK1, gel filtration | Mycobacterium tuberculosis |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Mycobacterium tuberculosis | - |
- |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| phosphoprotein | the protein is capable of autophosphorylation. Only His-491 is autophosphorylated and crucial for any enzymatic activity | Mycobacterium tuberculosis |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Ni-NTA column chromatography | Mycobacterium tuberculosis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ADP + (phosphate)20 | - |
Mycobacterium tuberculosis | ATP + (phosphate)19 | - |
r | |
| ATP + (phosphate)n | - |
Mycobacterium tuberculosis | ADP + (phosphate)n+1 | - |
r | |
| dADP + (phosphate)20 | - |
Mycobacterium tuberculosis | dATP + (phosphate)19 | - |
r | |
| GDP + (phosphate)20 | isoform PPK1 affinity for the ADP as a substrate is about 70fold high as opposed to GDP | Mycobacterium tuberculosis | GTP + (phosphate)19 | - |
r | |
| UDP + (phosphate)20 | - |
Mycobacterium tuberculosis | UTP + (phosphate)19 | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | 2 * 87000, isoform PPK1, SDS-PAGE | Mycobacterium tuberculosis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| polyphosphate kinase-1 | isoform | Mycobacterium tuberculosis |
| polyphosphate kinase-2 | isoform | Mycobacterium tuberculosis |
| PPK1 | isoform | Mycobacterium tuberculosis |
| PPK2 | isoform | Mycobacterium tuberculosis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 44.6 | - |
ATP | mutant H510Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 45 | - |
ATP | mutant H480Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 45.6 | - |
ATP | mutant H480A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 46.9 | - |
ATP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 55 | - |
ATP | mutant Y524A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 55.4 | - |
ATP | mutant H510A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 210 | - |
ADP | isoform PPK2, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 332 | - |
ADP | mutant H510A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 622 | - |
ADP | mutant H510Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 645 | - |
GDP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 943 | - |
ADP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 950 | - |
ADP | mutant H480A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 971 | - |
ADP | mutant H480Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 994 | - |
ADP | mutant Y524A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 4.2 | - |
ATP | mutant Y524A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 10.5 | - |
ATP | mutant H510A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 25.5 | - |
ADP | isoform PPK2, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 29 | - |
ATP | mutant H510Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 31 | - |
ATP | mutant H480A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 32.6 | - |
ATP | mutant H480Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 34.7 | - |
ATP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 49 | - |
ADP | mutant H510A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 57 | - |
GDP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 998 | - |
ADP | mutant Y524A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 3529 | - |
ADP | mutant H510Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 4690 | - |
ADP | mutant H480A of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 5222 | - |
ADP | mutant H480Q of isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
| 5743 | - |
ADP | isoform PPK1, pH and temperature not specified in the publication | Mycobacterium tuberculosis | |
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