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Literature summary extracted from

  • Misra, I.; Griffith, O.W.
    Expression and purification human g-glutamylcysteine synthetase (1998), Protein Expr. Purif., 13, 268-276.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.2 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

EC Number General Stability Organism
6.3.2.2 freezing of the purified recombinant enzyme in solution results in irreversible inactivation Homo sapiens
6.3.2.2 glycerol stabilizes Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.2.2 buthionine sulfoximine complete inhibition Homo sapiens
6.3.2.2 cysteamine complete inhibition Homo sapiens
6.3.2.2 GSH competitive feedback inhibition with respect to L-glutamate Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.2 0.1
-
 L-cysteine pH 8.2, 37°C Homo sapiens
6.3.2.2 0.4
-
 ATP pH 8.2, 37°C Homo sapiens
6.3.2.2 1.3
-
 4-aminobutyrate pH 8.2, 37°C Homo sapiens
6.3.2.2 1.8
-
 L-glutamate pH 8.2, 37°C Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.2 ATP + L-Glu + L-Cys Homo sapiens first and rate-limiting step in glutathione biosynthesis ADP + phosphate + gamma-L-Glu-L-Cys
-
 ir

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.2 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.2 recombinant holoenzyme from Escherichia coli, to homogeneity Homo sapiens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.2.2 25
-
 above, purified recombinant holoenzyme Homo sapiens

Storage Stability

EC Number Storage Stability Organism
6.3.2.2 -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
6.3.2.2 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.2 ATP + L-Glu + gamma-aminobutyrate
-
 Homo sapiens ADP + phosphate + L-Glu-gamma-aminobutyrate
-
 ir
6.3.2.2 ATP + L-Glu + L-Cys
-
 Homo sapiens ADP + phosphate + gamma-L-Glu-L-Cys
-
 ir
6.3.2.2 ATP + L-Glu + L-Cys first and rate-limiting step in glutathione biosynthesis Homo sapiens ADP + phosphate + gamma-L-Glu-L-Cys
-
 ir

Subunits

EC Number Subunits Comment Organism
6.3.2.2 More enzyme consists of 2 subunits, the heavy catalytic one and the light regulatory one Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
6.3.2.2 gamma-GCS
-
 Homo sapiens
6.3.2.2 gamma-Glutamylcysteine synthetase
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.2.2 37
-
 assay at Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.2 8.2
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.2.2 ATP dependent on Homo sapiens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.2.2 3.3
-
 GSH pH 8.2, 37°C Homo sapiens