Application | Comment | Organism |
---|---|---|
drug development | the enzym eis a target for inhibitor development | Francisella tularensis subsp. tularensis |
Cloned (Comment) | Organism |
---|---|
gene FTT1564, sequence comparisons, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21 Rosetta pLysS (DE3) | Francisella tularensis subsp. tularensis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, X-ray diffraction structure determination and analysis at 2.23 A resolution, molecular replacement using the Sinorhizobium meliloti PPK2 structure (SMc02148, PDB ID 3CZQ) as a model | Francisella tularensis subsp. tularensis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a DELTA FTT1564 enzyme gene deletion mutant strain | Francisella tularensis subsp. tularensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michalis-Menten steady state kinetic analysis of FtPPK2 substrate specificity, overview | Francisella tularensis subsp. tularensis | |
0.372 | - |
ATP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
0.546 | - |
ADP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
0.692 | - |
GTP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
0.727 | - |
GDP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, best at 10 mM | Francisella tularensis subsp. tularensis | |
Mn2+ | required, best at 1 mM | Francisella tularensis subsp. tularensis | |
additional information | lower optimal concentration for Mn2+ ions than Mg2+ ions | Francisella tularensis subsp. tularensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + (phosphate)n | Francisella tularensis subsp. tularensis | - |
ATP + (phosphate)n-1 | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Francisella tularensis subsp. tularensis | Q5NEQ5 | gene FTT1564 | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain BL21 Rosetta pLysS (DE3) by nickel affinity chromatography and gel filtration | Francisella tularensis subsp. tularensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + (phosphate)n | - |
Francisella tularensis subsp. tularensis | ATP + (phosphate)n-1 | - |
r | |
ATP + (phosphate)n+1 | - |
Francisella tularensis subsp. tularensis | ADP + (phosphate)n | - |
r | |
GDP + (phosphate)n | - |
Francisella tularensis subsp. tularensis | GTP + (phosphate)n-1 | - |
r | |
GTP + (phosphate)n+1 | - |
Francisella tularensis subsp. tularensis | GDP + (phosphate)n | - |
r | |
additional information | the substrate specificity of FtPPK2 includes purine but not pyrimidine nucleotides. No activity with UDP and CDP as substrates. No formation of ADP or GDP from AMP or GMP | Francisella tularensis subsp. tularensis | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the structure consists of a six-stranded parallel beta-sheet surrounded by 12 alpha-helices, with a high degree of similarity to other members of the PPK2 family and the thymidylate kinase superfamily | Francisella tularensis subsp. tularensis |
Synonyms | Comment | Organism |
---|---|---|
polyphosphate kinase 2 | - |
Francisella tularensis subsp. tularensis |
PPK2 | - |
Francisella tularensis subsp. tularensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Francisella tularensis subsp. tularensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.46 | - |
ATP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
2.77 | - |
GTP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
3.17 | - |
ADP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
3.69 | - |
GDP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Francisella tularensis subsp. tularensis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
maximal activity at pH 8.0, sharp drop-off in activity at pH 9 | Francisella tularensis subsp. tularensis |
General Information | Comment | Organism |
---|---|---|
evolution | a polyphosphate kinase from the polyphosphate kinase 2 (PPK2) family. The enzyme structure consists of a six-stranded parallelbeta-sheet surrounded by 12 alpha-helices, with a high degree of similarity to other members of the PPK2 family and the thymidylate kinase superfamily | Francisella tularensis subsp. tularensis |
malfunction | the isogenic deletion mutant is defective for intracellular growth in macrophages and is attenuated in mice. The DELTAFTT1564 strain shows significantly increased sensitivity to a range of antibiotics in a manner independent of the mode of action of the antibiotic | Francisella tularensis subsp. tularensis |
additional information | a lid-loop and the conserved Walker A and B motifs are important for substrate binding and enzyme catalysis | Francisella tularensis subsp. tularensis |
physiological function | important role for polyphosphate in the virulence of Francisella | Francisella tularensis subsp. tularensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.92 | - |
ATP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
4.002 | - |
GTP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
5.075 | - |
GDP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis | |
5.788 | - |
ADP | pH 8.0, 37°C | Francisella tularensis subsp. tularensis |