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Literature summary for 4.1.1.32 extracted from

  • Rios, S.E.; Nowak, T.
    Role of cysteine 306 in the catalytic mechanism of Ascaris suum phosphoenolpyruvate carboxykinase (2002), Arch. Biochem. Biophys., 404, 25-37.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
wild-type and mutant PEPCK, expression in Escherichia coli 1786 and JM109 Ascaris suum

Protein Variants

Protein Variants Comment Organism
C306A catalytically active mutant, kinetics Ascaris suum
C306S catalytically active mutant, kinetics Ascaris suum

Inhibitors

Inhibitors Comment Organism Structure
5,5'-dithiobis(2-nitrobenzoic acid) inhibits wild-type enzyme, but not C306A or C306S mutant enzymes Ascaris suum
additional information not inhibited by adenosine nucleotides Ascaris suum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.023
-
GDP pH 7.4, 25°C, wild-type enzyme Ascaris suum
0.052
-
GTP pH 7.4, 25°C, wild-type enzyme Ascaris suum
0.053
-
2'-deoxyguanosine 5'-diphosphate pH 7.4, 25°C, wild-type enzyme Ascaris suum
0.1
-
ITP pH 7.4, 25°C, wild-type enzyme Ascaris suum
0.11
-
ITP pH 7.4, 25°C, C306A mutant Ascaris suum
0.12
-
oxaloacetate pH 7.4, 25°C, wild-type enzyme Ascaris suum
0.12
-
GDP pH 7.4, 25°C, C306S mutant Ascaris suum
0.12
-
2'-deoxyguanosine 5'-diphosphate pH 7.4, 25°C, C306S mutant Ascaris suum
0.13
-
GTP pH 7.4, 25°C, C306S and C306A mutant Ascaris suum
0.15
-
GDP pH 7.4, 25°C, C306A mutant Ascaris suum
0.16
-
ITP pH 7.4, 25°C, C306S mutant Ascaris suum
0.19
-
2'-deoxyguanosine 5'-diphosphate pH 7.4, 25°C, C306A mutant Ascaris suum
0.19
-
oxaloacetate pH 7.4, 25°C, C306S mutant Ascaris suum
0.2
-
oxaloacetate pH 7.4, 25°C, C306A mutant Ascaris suum
0.3
-
IDP pH 7.4, 25°C, wild-type enzyme Ascaris suum
0.41
-
phosphoenolpyruvate pH 7.4, 25°C, C306S mutant Ascaris suum
0.48
-
phosphoenolpyruvate pH 7.4, 25°C, C306A mutant Ascaris suum
0.71
-
2'-deoxyguanosine 5'-triphosphate pH 7.4, 25°C, C306A mutant Ascaris suum
1.02
-
2'-deoxyguanosine 5'-triphosphate pH 7.4, 25°C, C306S mutant Ascaris suum
1.05
-
2'-deoxyguanosine 5'-triphosphate pH 7.4, 25°C, wild-type enzyme Ascaris suum
1.2
-
IDP pH 7.4, 25°C, C306A mutant Ascaris suum
1.38
-
IDP pH 7.4, 25°C, C306S mutant Ascaris suum
2.42
-
phosphoenolpyruvate pH 7.4, 25°C, wild-type enzyme Ascaris suum

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ enzyme binds Mn2+ at the catalytic site, binary PEPCK-Mn2+ complex, activator Mn2+ enhances the nucleotide binding Ascaris suum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
GTP + oxaloacetate Ascaris suum
-
GDP + phosphoenolpyruvate + CO2
-
?

Organism

Organism UniProt Comment Textmining
Ascaris suum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant and mutant enzyme Ascaris suum

Reaction

Reaction Comment Organism Reaction ID
GTP + oxaloacetate = GDP + phosphoenolpyruvate + CO2 catalytic mechanism Ascaris suum

Storage Stability

Storage Stability Organism
4°C, 3 weeks, stable Ascaris suum
room temperature, at least 6 h, stable Ascaris suum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2'-deoxyguanosine 5'-triphosphate + oxaloacetate 2’-deoxyGTP is a less effective substrate than GTP, 2’-deoxyGTP binds to enzyme less tight than 2’-deoxyGDP Ascaris suum 2'-deoxyguanosine 5'-diphosphate + phosphoenolpyruvate + CO2 2’-deoxyGDP is a less effective substrate than GDP in the reverse reaction ?
GTP + oxaloacetate
-
Ascaris suum GDP + phosphoenolpyruvate + CO2
-
?
GTP + oxaloacetate Cys-306 is important in nucleotide binding and may interact with the 2’-OH group in the ribose ring. GTP binds to enzyme tighter than GDP Ascaris suum GDP + phosphoenolpyruvate + CO2 GDP is a slightly more favorable substrate than IDP in the reverse reaction r
ITP + oxaloacetate
-
Ascaris suum IDP + phosphoenolpyruvate + CO2 GDP is a slightly more favorable substrate than IDP in the reverse reaction r
additional information not: adenosine nucleotides Ascaris suum ?
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Ascaris suum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.4
-
2'-deoxyguanosine 5'-diphosphate pH 7.4, 25°C, C306S mutant Ascaris suum
0.5
-
2'-deoxyguanosine 5'-diphosphate pH 7.4, 25°C, C306A mutant Ascaris suum
1.8
-
phosphoenolpyruvate pH 7.4, 25°C, C306A mutant Ascaris suum
2.1
-
phosphoenolpyruvate pH 7.4, 25°C, C306S mutant Ascaris suum
2.3
-
2'-deoxyguanosine 5'-triphosphate pH 7.4, 25°C, C306A mutant Ascaris suum
2.3
-
IDP pH 7.4, 25°C, C306S mutant Ascaris suum
2.4
-
IDP pH 7.4, 25°C, C306A mutant Ascaris suum
2.4
-
2'-deoxyguanosine 5'-triphosphate pH 7.4, 25°C, C306S mutant Ascaris suum
3.4
-
2'-deoxyguanosine 5'-triphosphate pH 7.4, 25°C, wild-type enzyme Ascaris suum
3.5
-
GDP pH 7.4, 25°C, C306S mutant Ascaris suum
4.1
-
GDP pH 7.4, 25°C, C306A mutant Ascaris suum
4.6
-
2'-deoxyguanosine 5'-diphosphate pH 7.4, 25°C, wild-type enzyme Ascaris suum
6.5
-
GDP pH 7.4, 25°C, wild-type enzyme Ascaris suum
6.9
-
IDP pH 7.4, 25°C, wild-type enzyme Ascaris suum
9.3
-
phosphoenolpyruvate pH 7.4, 25°C, wild-type enzyme Ascaris suum
10.3
-
GTP pH 7.4, 25°C, C306S mutant Ascaris suum
10.7
-
ITP pH 7.4, 25°C, C306A mutant Ascaris suum
12.4
-
GTP pH 7.4, 25°C, C306A mutant Ascaris suum
12.8
-
ITP pH 7.4, 25°C, C306S mutant Ascaris suum
13.3
-
oxaloacetate pH 7.4, 25°C, C306S mutant Ascaris suum
13.9
-
GTP pH 7.4, 25°C, wild-type enzyme Ascaris suum
14.5
-
oxaloacetate pH 7.4, 25°C, C306A mutant Ascaris suum
15.6
-
ITP pH 7.4, 25°C, wild-type enzyme Ascaris suum
17.8
-
oxaloacetate pH 7.4, 25°C, wild-type enzyme Ascaris suum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Ascaris suum