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Literature summary for 6.3.2.2 extracted from
- Expression and purification human g-glutamylcysteine synthetase (1998), Protein Expr. Purif., 13, 268-276.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| coexpression of the catalytic and the regulatory subunit from 2 different plasmids in Escherichia coli BL21(DE3), intracellular assembly of the holoenzyme | Homo sapiens |
General Stability
| General Stability | Organism |
|---|---|
| freezing of the purified recombinant enzyme in solution results in irreversible inactivation | Homo sapiens |
| glycerol stabilizes | Homo sapiens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| buthionine sulfoximine | complete inhibition | Homo sapiens |  |
| cysteamine | complete inhibition | Homo sapiens | |
| GSH | competitive feedback inhibition with respect to L-glutamate | Homo sapiens | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.1 | - |
L-cysteine | pH 8.2, 37°C | Homo sapiens | |
| 0.4 | - |
ATP | pH 8.2, 37°C | Homo sapiens | |
| 1.3 | - |
4-aminobutyrate | pH 8.2, 37°C | Homo sapiens | |
| 1.8 | - |
L-glutamate | pH 8.2, 37°C | Homo sapiens | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-Glu + L-Cys | Homo sapiens | first and rate-limiting step in glutathione biosynthesis | ADP + phosphate + gamma-L-Glu-L-Cys | - |
ir | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant holoenzyme from Escherichia coli, to homogeneity | Homo sapiens |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 25 | - |
above, purified recombinant holoenzyme | Homo sapiens |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -20°C, 25% glycerol, 20 mM imidazole HCl buffer, pH 7.4, 1 mM EDTA, without freezing, stable for at least 1 year | Homo sapiens |
| 4°C, purified recombinant enzyme, 20 mM imidazole HCl buffer, pH 7.4, 1 mM EDTA, stable for at least 7 days | Homo sapiens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-Glu + gamma-aminobutyrate | - |
Homo sapiens | ADP + phosphate + L-Glu-gamma-aminobutyrate | - |
ir | |
| ATP + L-Glu + L-Cys | - |
Homo sapiens | ADP + phosphate + gamma-L-Glu-L-Cys | - |
ir | |
| ATP + L-Glu + L-Cys | first and rate-limiting step in glutathione biosynthesis | Homo sapiens | ADP + phosphate + gamma-L-Glu-L-Cys | - |
ir | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | enzyme consists of 2 subunits, the heavy catalytic one and the light regulatory one | Homo sapiens |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| gamma-GCS | - |
Homo sapiens |
| gamma-Glutamylcysteine synthetase | - |
Homo sapiens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Homo sapiens |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.2 | - |
assay at | Homo sapiens |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | dependent on | Homo sapiens | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.3 | - |
GSH | pH 8.2, 37°C | Homo sapiens | |
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