Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Variovorax sp. |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapour diffusion method, enzyme in unbound state and in complex with Mg2+ and the inhibitor oxalate | Variovorax sp. |
Protein Variants | Comment | Organism |
---|---|---|
R188A | 1.15fold increase in turnover number | Variovorax sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | competitive | Variovorax sp. | |
oxalate | competitive | Variovorax sp. | |
phosphoenolpyruvate | competitive | Variovorax sp. | |
sulfopyruvate | competitive | Variovorax sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.005 | - |
3-phosphonopyruvate | 25°C, pH 7.5, wild-type enzyme | Variovorax sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates, Km: 0.0061 mM | Variovorax sp. | |
Mg2+ | activates, Km: 0.0035 mM. The Mg2+-binding is formed by the carboxylate groups of ASp54, Asp81, Asp83 and Glu110, with ASp81 coordinating directly to the metal, and the remaining carboxylate groups bridged by water molecules | Variovorax sp. | |
Mn2+ | activates, 0.00073 mM | Variovorax sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31000 | - |
4 * 31000, SDS-PAGE | Variovorax sp. |
110000 | - |
gel filtrration | Variovorax sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Variovorax sp. | Q84G06 | - |
- |
Variovorax sp. Pal2 | Q84G06 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant | Variovorax sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-phosphonopyruvate + H2O | rapid equilibrium ordered kinetic mechanism with Mg2+ binding first | Variovorax sp. | pyruvate + phosphate | - |
? | |
3-phosphonopyruvate + H2O | rapid equilibrium ordered kinetic mechanism with Mg2+ binding first | Variovorax sp. Pal2 | pyruvate + phosphate | - |
? | |
phosphoenolpyruvate + H2O | - |
Variovorax sp. | pyruvate + phosphate | - |
? | |
phosphoenolpyruvate + H2O | - |
Variovorax sp. Pal2 | pyruvate + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | 4 * 31000, SDS-PAGE | Variovorax sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
105 | - |
3-phosphonopyruvate | 25°C, pH 7.5, wild-type enzyme | Variovorax sp. | |
121 | - |
3-phosphonopyruvate | 25°C, pH 7.5, mutant enzyme R188A | Variovorax sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.017 | - |
sulfopyruvate | 25°C, pH 7.5 | Variovorax sp. | |
0.032 | - |
Ca2+ | 25°C, pH 7.5 | Variovorax sp. | |
0.21 | - |
phosphoenolpyruvate | 25°C, pH 7.5 | Variovorax sp. | |
2 | - |
oxalate | 25°C, pH 7.5 | Variovorax sp. |