Activating Compound | Comment | Organism | Structure |
---|---|---|---|
diphosphate | diphosphate binds Mg2+ to form a true substrate that activates the enzyme | Helicobacter pylori |
Cloned (Comment) | Organism |
---|---|
expression of His-tagged enzyme in Escherichia coli | Helicobacter pylori |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, free and in complex with diphosphate, hanging drop vapor diffusion method, for the free enzyme: 20 mg/ml protein in 20 mM Tris-HCl, pH 7.5, and 200 mM NaCl, precipitation with 1.44 M sodium citrate, and 100 mM Na HEPES, pH 7.4, 6 days at 25°C, for the diphosphate-bound enzyme: the diphosphate-PPase complex from the same buffer condition as the free enzyme with a 1:7.5 molar excess of K4diphosphate in the absence of divalent metal ion, the complex crystals are grown in the same buffer as free PPase using 3.6 M sodium formate as a precipitant, 3 days at 25°C, X-ray diffraction structure determination and analysis at 1.9 A and 2.3 A resolution, respectively, molecular replacement | Helicobacter pylori |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ATP | 50% inhibition at 2 mM, 90% at 3 mM ATP | Helicobacter pylori | |
imidodiphosphoric acid | a nonhydrolyzable PPi analogue, inhibits by 50% at 2 mM | Helicobacter pylori | |
N-ethylmaleimide | inhibits 38% of PPase activity at 37.5 mM | Helicobacter pylori | |
NaF | inhibits 46% and 91.4% of the activity at 0.04 mM and 0.5 mM, respectively | Helicobacter pylori |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Helicobacter pylori | |
0.214 | - |
diphosphate | recombinant enzyme | Helicobacter pylori |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Helicobacter pylori | 5829 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, binds to diphosphate, effects on enzyme activity and kinetics, pH-dependence, overview | Helicobacter pylori |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
20675 | - |
6 * 20675, recombinant His-tagged enzyme, mass spectrometry | Helicobacter pylori |
121000 | - |
recombinant His-tagged enzyme, gel filtration | Helicobacter pylori |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
diphosphate + H2O | Helicobacter pylori | PPase plays an essential role in energy conservation and provides the energy for many biosynthetic pathways controlling the intracellular diphosphate levels | 2 phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Helicobacter pylori | P56153 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli to homogeneity by nickel affinity chromatography and gel filtration | Helicobacter pylori |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
diphosphate + H2O | PPase plays an essential role in energy conservation and provides the energy for many biosynthetic pathways controlling the intracellular diphosphate levels | Helicobacter pylori | 2 phosphate | - |
? | |
diphosphate + H2O | diphosphate binding does not cause a significant conformational change in the diphosphate-PPase complex, structure, overview | Helicobacter pylori | 2 phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | 6 * 20675, recombinant His-tagged enzyme, mass spectrometry | Helicobacter pylori |
More | structure analysis, PPase comprises three alpha-helices and nine beta-strands and folds as a barrel structure, it forms a hexamer in both the solution and crystal states, and each monomer has its own PPi-binding site | Helicobacter pylori |
Synonyms | Comment | Organism |
---|---|---|
inorganic pyrophosphatase | - |
Helicobacter pylori |
PPase | - |
Helicobacter pylori |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Helicobacter pylori |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Helicobacter pylori |