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Literature summary extracted from

  • Johnson, T.A.; Holyoak, T.
    Increasing the conformational entropy of the Omega-loop lid domain in phosphoenolpyruvate carboxykinase impairs catalysis and decreases catalytic fidelity (2010), Biochemistry, 49, 5176-5187.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.1.32 expressed in Escherichia coli BL21(DE3) cells Rattus norvegicus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.1.1.32 mutant enzyme A467G in complex with Mn2+, substrates and inhibitors Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
4.1.1.32 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

EC Number Inhibitors Comment Organism Structure
4.1.1.32 2-Phosphoglycolate
-
 Rattus norvegicus
4.1.1.32 beta-sulfopyruvate mixed inhibitor of PEPCK against oxaloacetate Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.1.32 0.039
-
 GDP wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 0.047
-
 GTP mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 0.052
-
 oxaloacetate wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 0.063
-
 phosphoenolpyruvate mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 0.068
-
 GTP wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 0.07
-
 GDP mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 0.294
-
 phosphoenolpyruvate wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 0.749
-
 oxaloacetate mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 0.814
-
 CO2 mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 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

EC Number Localization Comment Organism GeneOntology No. Textmining
4.1.1.32 cytosol
-
 Rattus norvegicus 5829
-

Organism

EC Number Organism UniProt Comment Textmining
4.1.1.32 Rattus norvegicus P07379
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.1.32
-
 Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.1.32 GDP + phosphoenolpyruvate + CO2
-
 Rattus norvegicus GTP + oxaloacetate
-
 r
4.1.1.32 GTP + oxaloacetate physiological direction of reaction Rattus norvegicus GDP + phosphoenolpyruvate + CO2
-
 r

Synonyms

EC Number Synonyms Comment Organism
4.1.1.32 PEPCK
-
 Rattus norvegicus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.1.32 1
-
 CO2 mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 1
-
 phosphoenolpyruvate mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 1
-
 GDP mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 14
-
 GTP mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 14
-
 oxaloacetate mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 19
-
 CO2 wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 19
-
 phosphoenolpyruvate wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 19
-
 GDP wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 54
-
 GTP wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 54
-
 oxaloacetate wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.1.32 GTP
-
 Rattus norvegicus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.1.1.32 1.2
-
 CO2 mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 14
-
 GDP mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 15
-
 phosphoenolpyruvate mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 16
-
 CO2 wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 19
-
 oxaloacetate mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 66
-
 phosphoenolpyruvate wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 270
-
 GTP mutant enzyme A467G, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 510
-
 GDP wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 790
-
 GTP wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus
4.1.1.32 1000
-
 oxaloacetate wild type enzyme, in 50 mM HEPES pH 7.5, 10 mM dithiothreitol, 4 mM MgCl2, at 25°C Rattus norvegicus