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Literature summary for 6.3.2.3 extracted from
- The glutathione synthetase of Schizosaccharomyces pombe is synthesized as a homodimer but retains full activity when present as a heterotetramer (2003), J. Biol. Chem., 278, 40152-40161.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| DNA sequence analysis, overexpression of gene gshs2 in an enzyme-deficient Schizosaccharomyces pombe strain, expression of wild-type enzyme, isolated enzyme subunit fragments and mutants as C-terminally His-tagged proteins, subcloning in Escherichia coli DH5alpha | Schizosaccharomyces pombe |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| structure modeling | Schizosaccharomyces pombe |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | elimination of the protease cleavage site in the 56 kDa subunit by site-directed mutagenesis results in expression of a stable and functional enzyme | Schizosaccharomyces pombe |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 56000 | - |
2 * 56000, SDS-PAGE | Schizosaccharomyces pombe |
| 120000 | - |
gel filtration | Schizosaccharomyces pombe |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Schizosaccharomyces pombe | - |
gene gshs2 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes from Schizosaccharomyces pombe mutant strain, to homogeneity | Schizosaccharomyces pombe |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 17.2 | - |
purified recombinant wild-type enzyme after complete proteoltic cleavage | Schizosaccharomyces pombe |
| 17.3 | - |
purified recombinant isolated subfragments of 24 and 36 kDa | Schizosaccharomyces pombe |
| 19.3 | - |
purified recombinant deletion mutant lacking 15 amino acids | Schizosaccharomyces pombe |
| 20.1 | - |
purified recombinant wild-type enzyme | Schizosaccharomyces pombe |
| 21.9 | - |
purified recombinant enzyme mutant with eliminated cleavage site | Schizosaccharomyces pombe |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + gamma-L-Glu-L-Cys + Gly | cells are not able to grow without glutathione, wild-type and all mutant enzyme forms can restore activity of the deficient mutant strain on minimal medium without glutathione | Schizosaccharomyces pombe | ADP + phosphate + glutathione | - |
? |  |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 56000, SDS-PAGE | Schizosaccharomyces pombe |
| More | identification of the protease cleavage site of the 56 kDa subunit, which is cleaved into a 24 kDa and a 36 kDa fragment, also on storage at -20°C, the formed heterotetramer is fully active | Schizosaccharomyces pombe |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Glutathione synthetase | - |
Schizosaccharomyces pombe |
| GS | - |
Schizosaccharomyces pombe |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.2 | - |
assay at | Schizosaccharomyces pombe |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | - |
Schizosaccharomyces pombe | |
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Release 2023.1 (January 2023)
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