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Literature summary extracted from

  • Phlippen, N.; Hoffmann, K.; Fischer, R.; Wolf, K.; Zimmermann, M.
    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.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.3 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)

EC Number Crystallization (Comment) Organism
6.3.2.3 structure modeling Schizosaccharomyces pombe

Protein Variants

EC Number Protein Variants Comment Organism
6.3.2.3 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.2.3 56000
-
 2 * 56000, SDS-PAGE Schizosaccharomyces pombe
6.3.2.3 120000
-
 gel filtration Schizosaccharomyces pombe

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.3 Schizosaccharomyces pombe
-
 gene gshs2
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.3 recombinant His-tagged wild-type and mutant enzymes from Schizosaccharomyces pombe mutant strain, to homogeneity Schizosaccharomyces pombe

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.2.3 17.2
-
 purified recombinant wild-type enzyme after complete proteoltic cleavage Schizosaccharomyces pombe
6.3.2.3 17.3
-
 purified recombinant isolated subfragments of 24 and 36 kDa Schizosaccharomyces pombe
6.3.2.3 19.3
-
 purified recombinant deletion mutant lacking 15 amino acids Schizosaccharomyces pombe
6.3.2.3 20.1
-
 purified recombinant wild-type enzyme Schizosaccharomyces pombe
6.3.2.3 21.9
-
 purified recombinant enzyme mutant with eliminated cleavage site Schizosaccharomyces pombe

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.3 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

EC Number Subunits Comment Organism
6.3.2.3 dimer 2 * 56000, SDS-PAGE Schizosaccharomyces pombe
6.3.2.3 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

EC Number Synonyms Comment Organism
6.3.2.3 Glutathione synthetase
-
 Schizosaccharomyces pombe
6.3.2.3 GS
-
 Schizosaccharomyces pombe

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