Protein Variants | Comment | Organism |
---|---|---|
additional information | naturally occuring ehPPase mutant containing an inherent mutation in an otherwise conserved asparagine residue in a loop near the active site exhibits noncooperative hydrolysis kinetics | Ethanoligenens harbinense |
additional information | naturally occuring ehPPase mutant containing an inherent mutation in an otherwise conserved asparagine residue in a loop near the active site exhibits noncooperative hydrolysis kinetics | Desulfitobacterium hafniense |
N312S | site-directed mutagenesis, the mutant shows altered Mg2+ dependence and kinetics compared to wild-type. The N312S substitution in dhPPase abolishes kinetic cooperativity and reverses the effect of Ap4A on it. In the presence of 0.05 mMATP and 5 mM Mg2+, the hydrolysis kinetics of N312S-dhPPase remains non-cooperative, the catalytic constant increases 1.5fold, and the average Km value increases about 5.5fold | Desulfitobacterium hafniense |
S213N | site-directed mutagenesis, the substitution confers significant negative cooperativity on the enzyme, decreasing the Hill coefficient to 0.7, and the Km2/Km1 ratio increases to approximately 13 over the entire range of Mg2+ concentrations. The catalytic constant decreases by a factor of 4.5 upon Ser213 replacement | Ethanoligenens harbinense |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | ehPPase lacks kinetic cooperativity, kinetic analysis, Michaelis-Menten kinetics, overview. The S213N substitution conferred significant negative cooperativity on the enzyme, decreasing the Hill coefficient to 0.7, and the Km2/Km1 ratio increases to approximately 13 over the entire range of Mg2+ concentrations | Ethanoligenens harbinense | |
additional information | - |
additional information | kinetic analysis, Michaelis-Menten kinetics, overview. With the wild-type enzyme, hydrolysis kinetics remains positively cooperative in the presence of ATP | Desulfitobacterium hafniense | |
0.049 | 0.097 | diphosphate | pH 7.2, 25°C, mutant enzyme N312S | Desulfitobacterium hafniense | |
0.6 | - |
diphosphate | pH 7.2, 25°C, wild-type enzyme | Desulfitobacterium hafniense | |
35 | - |
diphosphate | pH 7.2, 25°C, mutant enzyme S213N | Ethanoligenens harbinense | |
46 | - |
diphosphate | pH 7.2, 25°C, wild-type enzyme | Ethanoligenens harbinense |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
soluble | - |
Ethanoligenens harbinense | - |
- |
soluble | - |
Desulfitobacterium hafniense | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | required | Ethanoligenens harbinense | |
Co2+ | required | Desulfitobacterium hafniense | |
Mg2+ | dependent on | Desulfitobacterium hafniense | |
Mg2+ | dependent on, Mg2+ concentration-dependence of kinetic cooperativity in wild-type ehPPase and mutant S213N-ehPPase, overview | Ethanoligenens harbinense | |
Mn2+ | required | Ethanoligenens harbinense | |
Mn2+ | required | Desulfitobacterium hafniense |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
diphosphate + H2O | Ethanoligenens harbinense | - |
2 phosphate | - |
r | |
diphosphate + H2O | Desulfitobacterium hafniense | - |
2 phosphate | - |
r | |
diphosphate + H2O | Desulfitobacterium hafniense DSM 10664 | - |
2 phosphate | - |
r | |
diphosphate + H2O | Desulfitobacterium hafniense DCB-2 | - |
2 phosphate | - |
r | |
diphosphate + H2O | Ethanoligenens harbinense UniProt | - |
2 phosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Desulfitobacterium hafniense | B8FP42 | - |
- |
Desulfitobacterium hafniense DCB-2 | B8FP42 | - |
- |
Desulfitobacterium hafniense DSM 10664 | B8FP42 | - |
- |
Ethanoligenens harbinense | E6U5H4 | - |
- |
Ethanoligenens harbinense UniProt | E6U5H4 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
diphosphate + H2O | - |
Ethanoligenens harbinense | 2 phosphate | - |
r | |
diphosphate + H2O | - |
Desulfitobacterium hafniense | 2 phosphate | - |
r | |
diphosphate + H2O | - |
Desulfitobacterium hafniense DSM 10664 | 2 phosphate | - |
r | |
diphosphate + H2O | - |
Desulfitobacterium hafniense DCB-2 | 2 phosphate | - |
r | |
diphosphate + H2O | - |
Ethanoligenens harbinense UniProt | 2 phosphate | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | - |
Ethanoligenens harbinense |
homodimer | - |
Desulfitobacterium hafniense |
Synonyms | Comment | Organism |
---|---|---|
dhPPase | - |
Desulfitobacterium hafniense |
ehPPase | - |
Ethanoligenens harbinense |
nucleotide-regulated pyrophosphatase | - |
Ethanoligenens harbinense |
nucleotide-regulated pyrophosphatase | - |
Desulfitobacterium hafniense |
PPase | - |
Ethanoligenens harbinense |
PPase | - |
Desulfitobacterium hafniense |
pyrophosphatase | - |
Ethanoligenens harbinense |
pyrophosphatase | - |
Desulfitobacterium hafniense |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Ethanoligenens harbinense |
25 | - |
assay at | Desulfitobacterium hafniense |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
10 | - |
diphosphate | pH 7.2, 25°C, mutant enzyme S213N | Desulfitobacterium hafniense | |
320 | - |
diphosphate | pH 7.2, 25°C, wild-type enzyme | Desulfitobacterium hafniense | |
410 | - |
diphosphate | pH 7.2, 25°C, mutant enzyme S213N | Ethanoligenens harbinense | |
1850 | - |
diphosphate | pH 7.2, 25°C, wild-type enzyme | Ethanoligenens harbinense |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Ethanoligenens harbinense |
7.2 | - |
assay at | Desulfitobacterium hafniense |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the CBS-PPases | Ethanoligenens harbinense |
evolution | the enzyme belongs to the CBS-PPases | Desulfitobacterium hafniense |
malfunction | naturally occuring ehPPase mutant containing an inherent mutation in an otherwise conserved asparagine residue in a loop near the active site exhibits noncooperative hydrolysis kinetics | Ethanoligenens harbinense |
malfunction | naturally occuring ehPPase mutant containing an inherent mutation in an otherwise conserved asparagine residue in a loop near the active site exhibits noncooperative hydrolysis kinetics | Desulfitobacterium hafniense |
additional information | comparison of the primary structure of ehPPase with those of five other CBS-PPases. Molecular dynamic simulations, overview | Ethanoligenens harbinense |
additional information | structure comparisons of CBS-PPases and molecular dynamic simulations, overview | Desulfitobacterium hafniense |
physiological function | ehPPase lacks kinetic cooperativity and is not regulated by adenine nucleotides. ehPPase shows insensitivity (below 10% activity change) to adenine nucleotides (AMP, ADP, ATP and diadenosine polyphosphates, ApnA, with n=3-6) over a wide range of substrate concentrations (0.001-0.30 mM), metal cofactor concentrations (0.05-20 mM), and nucleotide concentrations (10 nM-1.0 mM) for mononucleotides and 0.01 nM-0.1 mM for dinucleotides | Ethanoligenens harbinense |