Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 6.3.2.3 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
Search for more literature for 6.3.2.3

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
D458A site-directed mutagenesis, the mutant shows 10% activity compared to the wild-type enzyme Homo sapiens
D458N site-directed mutagenesis, the mutant shows 15% activity compared to the wild-type enzyme Homo sapiens
D458R site-directed mutagenesis, the mutant shows 7% activity compared to the wild-type enzyme Homo sapiens
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]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Homo sapiens

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Homo sapiens P48637
-
-

Substrates and Products (Substrate)

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Homo sapiens

General Information

General Information Comment Organism
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