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Literature summary for 4.1.1.32 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
Search for more literature for 4.1.1.32

Cloned(Commentary)

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

Crystallization (Commentary)

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

Protein Variants

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

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

KM Value [mM]

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

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

Organism

Organism UniProt Comment Textmining
Rattus norvegicus P07379
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Rattus norvegicus

Substrates and Products (Substrate)

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

Synonyms Comment Organism
PEPCK
-
 Rattus norvegicus

Turnover Number [1/s]

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

Cofactor Comment Organism Structure
GTP
-
 Rattus norvegicus

kcat/KM [mM/s]

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