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 |
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 |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
6.3.2.3 | Mg2+ | required | Schizosaccharomyces pombe |
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 |
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 | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
6.3.2.3 | Schizosaccharomyces pombe | P35669 | - |
- |
6.3.2.23 | Medicago truncatula | - |
- |
- |
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 |
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 | ? | - |
? |
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 |
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 |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
6.3.2.3 | 8.2 | - |
assay at | Schizosaccharomyces pombe |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
6.3.2.3 | ATP | - |
Schizosaccharomyces pombe | |
6.3.2.23 | ATP | - |
Medicago truncatula |
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 |