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Literature summary extracted from

  • Brown, T.R.; Drummond, M.L.; Barelier, S.; Crutchfield, A.S.; Dinescu, A.; Slavens, K.D.; Cundari, T.R.; Anderson, M.E.
    Aspartate 458 of human glutathione synthetase is important for cooperativity and active site structure (2011), Biochem. Biophys. Res. Commun., 411, 536-542.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.3 gene gshB, expression of His-tagged wild-type and mutants Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
6.3.2.3 D458A site-directed mutagenesis, the mutant shows 10% activity compared to the wild-type enzyme Homo sapiens
6.3.2.3 D458N site-directed mutagenesis, the mutant shows 15% activity compared to the wild-type enzyme Homo sapiens
6.3.2.3 D458R site-directed mutagenesis, the mutant shows 7% activity compared to the wild-type enzyme Homo sapiens
6.3.2.3 additional information all Asp458 mutants display a change in cooperativity from negative cooperativity to non-cooperative. All mutants show similar stability as compared to wild-type enzyme, differential scanning calorimetry Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.3 0.1 2 gamma-L-glutamyl-L-cysteine pH 8.2, 37°C, D458N Homo sapiens
6.3.2.3 0.25
-
 gamma-L-glutamyl-L-cysteine pH 8.2, 37°C, D458A Homo sapiens
6.3.2.3 0.29
-
 gamma-L-glutamyl-L-cysteine pH 8.2, 37°C, D458R Homo sapiens
6.3.2.3 0.58
-
 glycine pH 8.2, 37°C, wild-type enzyme Homo sapiens
6.3.2.3 1.41
-
 gamma-L-glutamyl-L-cysteine pH 8.2, 37°C, wild-type enzyme Homo sapiens
6.3.2.3 3 5 glycine pH 8.2, 37°C, D458A Homo sapiens
6.3.2.3 16
-
 glycine pH 8.2, 37°C, D458N Homo sapiens
6.3.2.3 51
-
 glycine pH 8.2, 37°C, D458R Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.2.3 Mg2+ required Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.3 ATP + gamma-L-glutamyl-L-cysteine + glycine Homo sapiens
-
 ADP + phosphate + glutathione
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.3 Homo sapiens P48637
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.3 ATP + gamma-L-glutamyl-L-cysteine + glycine
-
 Homo sapiens ADP + phosphate + glutathione
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.2.3 37
-
 assay at Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.2.3 1.17
-
 glycine pH 8.2, 37°C, D458R Homo sapiens
6.3.2.3 1.17
-
 gamma-L-glutamyl-L-cysteine pH 8.2, 37°C, D458R Homo sapiens
6.3.2.3 1.6
-
 glycine pH 8.2, 37°C, D458A Homo sapiens
6.3.2.3 1.6
-
 gamma-L-glutamyl-L-cysteine pH 8.2, 37°C, D458A Homo sapiens
6.3.2.3 2.31
-
 glycine pH 8.2, 37°C, D458N Homo sapiens
6.3.2.3 2.31
-
 gamma-L-glutamyl-L-cysteine pH 8.2, 37°C, D458N Homo sapiens
6.3.2.3 15.68
-
 glycine pH 8.2, 37°C, wild-type enzyme Homo sapiens
6.3.2.3 15.68
-
 gamma-L-glutamyl-L-cysteine pH 8.2, 37°C, wild-type enzyme Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.3 8.2
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.2.3 ATP
-
 Homo sapiens

General Information

EC Number General Information Comment Organism
6.3.2.3 additional information the active site is composed of three highly conserved catalytic loops, notably the alanine rich A-loop, Asp458 is important for cooperativity and active site structure, it impacts the allostery of the enzyme. Asp458 is important for loop closure and is a critical residue within the enzyme's A-loop. Enzyme structure-function modeling, molecular dynamics, overview Homo sapiens