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Literature summary extracted from

  • Dworeck, T.; Zimmermann, M.
    Site directed mutagenesis of Schizosaccharomyces pombe glutathione synthetase produces an enzyme with homoglutathione synthetase activity (2012), PLoS ONE, 7, e46580.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.3 sequence comparison, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain DH5alpha Schizosaccharomyces pombe

Protein Variants

EC Number Protein Variants Comment Organism
6.3.2.3 A485L/T486P site-directed mutagenesis, the mutant shows 70% reduced activity compared to the wild-type activity, and a shift of substrate specificity with increased affinity of GSH2 for Ser as a substrate, while affinity to Gly is preserved. This provides another biosynthetic pathway for hydroxymethyl glutathione, which is known to be synthesized from glutathione and Ser in a reaction catalysed by carboxypeptidase Y Schizosaccharomyces pombe
6.3.2.3 I471M/C472M/A485L/T486P site-directed mutagenesis, the mutant shows 38% reduced activity compared to the wild-type activity, and a shift of substrate specificity with 1.2fold increased affinity of GSH2 for beta-Ala and lowered affinity for Gly, which is a characteristic of the enzyme homoglutathione synthetase found in plants, EC 6.3.2.23 Schizosaccharomyces pombe
6.3.2.3 I471M/C472V site-directed mutagenesis, the mutant shows showed much lower affinity towards Gly and 78% reduced activity compared to the wild-type activity, but no other differences Schizosaccharomyces pombe
6.3.2.23 L534A/P535A site-directed mutagenesis, the mutant shows properties similar to GSH synthetase, EC 6.3.2.3. A534 and A535 are conserved in glutathione synthetase Medicago truncatula

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.2.3 Mg2+ required Schizosaccharomyces pombe

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.2.3 56000
-
x * 56000, about, recombinant His-tagged wild-type and mutant enzymes, SDS-PAGE Schizosaccharomyces pombe

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 Schizosaccharomyces pombe
-
ADP + phosphate + glutathione
-
?
6.3.2.23 ATP + gamma-L-glutamyl-L-cysteine + beta-alanine Medicago truncatula
-
ADP + phosphate + gamma-L-glutamyl-L-cysteinyl-beta-alanine
-
?
6.3.2.23 ATP + gamma-L-glutamyl-L-cysteine + L-serine Medicago truncatula
-
ADP + phosphate + gamma-L-glutamyl-L-cysteinyl-L-serine
-
?
6.3.2.23 additional information Medicago truncatula hGSH synthetase found in plants is much more specific for beta-Ala, although it also accepts Gly as a substrate ?
-
?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.3 Schizosaccharomyces pombe P35669
-
-
6.3.2.23 Medicago truncatula
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.3 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain DH5alpha by nickel affinity chromatography, gel filtration, and Cibacron blue 3GA affinity chromatography Schizosaccharomyces pombe

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
-
Schizosaccharomyces pombe ADP + phosphate + glutathione
-
?
6.3.2.3 additional information no activity with beta-alanine and L-serine by wild-type glutathione synthetase Schizosaccharomyces pombe ?
-
?
6.3.2.23 ATP + gamma-L-glutamyl-L-cysteine + beta-alanine
-
Medicago truncatula ADP + phosphate + gamma-L-glutamyl-L-cysteinyl-beta-alanine
-
?
6.3.2.23 ATP + gamma-L-glutamyl-L-cysteine + beta-alanine
-
Medicago truncatula ADP + phosphate + gamma-L-glutamyl-L-cysteinyl-beta-alanine i.e. homo-glutathione ?
6.3.2.23 ATP + gamma-L-glutamyl-L-cysteine + L-serine
-
Medicago truncatula ADP + phosphate + gamma-L-glutamyl-L-cysteinyl-L-serine
-
?
6.3.2.23 ATP + gamma-L-glutamyl-L-cysteine + L-serine
-
Medicago truncatula ADP + phosphate + gamma-L-glutamyl-L-cysteinyl-L-serine i.e. hydroxymethyl-glutathione ?
6.3.2.23 additional information hGSH synthetase found in plants is much more specific for beta-Ala, although it also accepts Gly as a substrate Medicago truncatula ?
-
?

Subunits

EC Number Subunits Comment Organism
6.3.2.3 ? x * 56000, about, recombinant His-tagged wild-type and mutant enzymes, SDS-PAGE Schizosaccharomyces pombe
6.3.2.3 More structure comparisons and homology structure molecular modelling of the GSH2 wild-type and mutant enzymes, overview Schizosaccharomyces pombe

Synonyms

EC Number Synonyms Comment Organism
6.3.2.3 Glutathione synthetase
-
Schizosaccharomyces pombe
6.3.2.3 GSH2
-
Schizosaccharomyces pombe
6.3.2.23 hGSH synthetase
-
Medicago truncatula
6.3.2.23 hGSH-synthetase
-
Medicago truncatula
6.3.2.23 homoglutathione synthetase
-
Medicago truncatula

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.3 8.2
-
assay at Schizosaccharomyces pombe

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.2.3 ATP
-
Schizosaccharomyces pombe
6.3.2.23 ATP
-
Medicago truncatula

General Information

EC Number General Information Comment Organism
6.3.2.3 evolution homoglutathione synthetase, EC 6.3.2.23, has evolved from glutathione synthetase by a single gene duplication event Schizosaccharomyces pombe
6.3.2.3 metabolism A485L/T486P mutant shows a shift the substrate specificity increased affinity of GSH2 for Ser as a substrate, while affinity to Gly is preserved. This provides a new biosynthetic pathway for hydroxymethyl glutathione, which is known to be synthesized from glutathione and Ser in a reaction catalysed by carboxypeptidase Y Schizosaccharomyces pombe
6.3.2.3 additional information structure comparisons and homology structure molecular modelling of the GSH2 wild-type and mutant enzymes, overview Schizosaccharomyces pombe
6.3.2.23 evolution homoglutathione synthetase has evolved from glutathione synthetase, EC 6.3.2.3, by a single gene duplication event Medicago truncatula