Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Rattus norvegicus |
Crystallization (Comment) | Organism |
---|---|
mutant enzyme A467G in complex with Mn2+, substrates and inhibitors | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
A467G | PEPCK has a 14fold higher Km value for oxaloacetate than wild type, coupled with a reduction in kcat (26% of wild type), resulting in a reduction in catalytic efficiency by nearly two orders of magnitude (1.9% of wild type). There is little change in the Km for GTP (factor of 1.4), resulting in the catalytic efficiency for GTP decreasing by less than a factor of three. In the reverse reaction, the mutant shows a decrease in the Km value for phosphoenolpyruvate (21% of wild type), a kcat reduced to 5% of the wild type value, and a factor of four reduction in the catalytic efficiency relative to wild type | Rattus norvegicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-Phosphoglycolate | - |
Rattus norvegicus | |
beta-sulfopyruvate | mixed inhibitor of PEPCK against oxaloacetate | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.039 | - |
GDP | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
0.047 | - |
GTP | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
0.052 | - |
oxaloacetate | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
0.063 | - |
phosphoenolpyruvate | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
0.068 | - |
GTP | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
0.07 | - |
GDP | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
0.294 | - |
phosphoenolpyruvate | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
0.749 | - |
oxaloacetate | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
0.814 | - |
CO2 | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
1.194 | - |
CO2 | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Rattus norvegicus | 5829 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | P07379 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GDP + phosphoenolpyruvate + CO2 | - |
Rattus norvegicus | GTP + oxaloacetate | - |
r | |
GTP + oxaloacetate | physiological direction of reaction | Rattus norvegicus | GDP + phosphoenolpyruvate + CO2 | - |
r |
Synonyms | Comment | Organism |
---|---|---|
PEPCK | - |
Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1 | - |
CO2 | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
1 | - |
phosphoenolpyruvate | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
1 | - |
GDP | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
14 | - |
GTP | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
14 | - |
oxaloacetate | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
19 | - |
CO2 | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
19 | - |
phosphoenolpyruvate | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
19 | - |
GDP | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
54 | - |
GTP | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
54 | - |
oxaloacetate | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
GTP | - |
Rattus norvegicus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.2 | - |
CO2 | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
14 | - |
GDP | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
15 | - |
phosphoenolpyruvate | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
16 | - |
CO2 | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
19 | - |
oxaloacetate | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
66 | - |
phosphoenolpyruvate | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
270 | - |
GTP | mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
510 | - |
GDP | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
790 | - |
GTP | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus | |
1000 | - |
oxaloacetate | wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C | Rattus norvegicus |