Cloned (Comment) | Organism |
---|---|
wild-type and mutant PEPCK, expression in Escherichia coli 1786 and JM109 | Ascaris suum |
Protein Variants | Comment | Organism |
---|---|---|
C306A | catalytically active mutant, kinetics | Ascaris suum |
C306S | catalytically active mutant, kinetics | Ascaris suum |
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 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 | 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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + oxaloacetate | Ascaris suum | - |
GDP + phosphoenolpyruvate + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ascaris suum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant and mutant enzyme | Ascaris suum |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
GTP + oxaloacetate = GDP + phosphoenolpyruvate + CO2 | catalytic mechanism | Ascaris suum |
Storage Stability | Organism |
---|---|
4°C, 3 weeks, stable | Ascaris suum |
room temperature, at least 6 h, stable | Ascaris suum |
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 Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Ascaris suum |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Ascaris suum |