Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (DE3) cells | Pseudomonas aeruginosa |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapour diffusion method, aliquots of 10 mg/ml protein in 5 mM MgCl2, 5 mM phosphonopyruvate and 10 mM NaHEPES (pH 7.0) mixed with equal volumes of reservoir solution containing 12% polyethylene glycol 20000 and 0.1 M MES (pH 6.0) | Pseudomonas aeruginosa |
Protein Variants | Comment | Organism |
---|---|---|
H235A | the kcat value for catalysis of oxaloacetate decarboxylation is less than an order of magnitude smaller compared to the wild type enzyme | Pseudomonas aeruginosa |
H235Q | the kcat value for catalysis of oxaloacetate decarboxylation is less than an order of magnitude smaller compared to the wild type enzyme | Pseudomonas aeruginosa |
Y212F | 25fold reduction of the Km value compared to the wild type enzyme | Pseudomonas aeruginosa |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3,3-difluoroxaloacetate | - |
Pseudomonas aeruginosa | |
Acetopyruvate | - |
Pseudomonas aeruginosa | |
alpha-ketovalerate | - |
Pseudomonas aeruginosa | |
oxalate | - |
Pseudomonas aeruginosa | |
Phosphonopyruvate | - |
Pseudomonas aeruginosa | |
pyruvate | - |
Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.087 | - |
oxaloacetate | mutant enzyme Y212F, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C | Pseudomonas aeruginosa | |
0.63 | - |
3-methyloxaloacetate | wild type enzyme, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C | Pseudomonas aeruginosa | |
1.02 | - |
oxaloacetate | mutant enzyme H235A, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C | Pseudomonas aeruginosa | |
2.2 | - |
oxaloacetate | wild type enzyme, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C | Pseudomonas aeruginosa | |
2.5 | - |
oxaloacetate | mutant enzyme H235Q, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C | Pseudomonas aeruginosa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | cofactor | Pseudomonas aeruginosa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | Q9HUU1 | - |
- |
Purification (Comment) | Organism |
---|---|
DEAE-cellulose column chromatography, phenyl-Sepharose columnn chromatography, and butyl-Sepharose column chromatography | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-methyloxaloacetate | - |
Pseudomonas aeruginosa | 2-oxobutanoate + CO2 | - |
? | |
Oxaloacetate | - |
Pseudomonas aeruginosa | Pyruvate + CO2 | - |
r |
Subunits | Comment | Organism |
---|---|---|
dimer | dimer of dimers, X-ray crystallography | Pseudomonas aeruginosa |
Synonyms | Comment | Organism |
---|---|---|
PA4872 | member of the PEP mutase/isocitrate lyase superfamily | Pseudomonas aeruginosa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
250 | - |
3-methyloxaloacetate | in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C | Pseudomonas aeruginosa | |
7500 | - |
oxaloacetate | in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C | Pseudomonas aeruginosa |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.043 | - |
oxalate | - |
Pseudomonas aeruginosa | |
0.45 | - |
3,3-difluoroxaloacetate | - |
Pseudomonas aeruginosa | |
1.09 | - |
Acetopyruvate | - |
Pseudomonas aeruginosa | |
3 | - |
Phosphonopyruvate | - |
Pseudomonas aeruginosa | |
6.7 | - |
alpha-ketovalerate | - |
Pseudomonas aeruginosa | |
7.2 | - |
pyruvate | - |
Pseudomonas aeruginosa |