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Information on EC 6.3.2.3 - glutathione synthase and Organism(s) Schizosaccharomyces pombe and UniProt Accession P35669
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6.3.2.3 glutathione synthaseSpecify your search results
Word Map
- 6.3.2.3
- gamma-glutamylcysteine
- dismutase
-
gamma-glutamyl
- catalase
- s-transferase
- sulfoximine
- cadmium
-
5-oxoprolinuria
-
buthionine
- 5-oxoproline
-
hemolytic
- acidosis
- phytochelatins
- transpeptidase
- gamma-gcs
-
school
-
school-based
-
pyroglutamic
-
glutathione-related
-
atp-grasp
-
bullied
-
l-buthionine-s,r-sulfoximine
- juncea
-
6.3.2.2
-
glutathione-deficient
-
homoglutathione
-
gsh-dependent
- medicine
- biotechnology
- synthesis
- analysis
The taxonomic range for the selected organisms is: Schizosaccharomyces pombe
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
glutathione synthetase, glutathione synthase, gsh synthetase, gshs, gsh-s, gsh synthase, tags2, gshii, gshs1, tags1, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Glutathione synthase
-
-
-
-
Glutathione synthetase
Glutathione synthetase (tripeptide)
-
-
-
-
GSH synthetase
-
-
-
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GSHase
-
-
-
-
GSS
-
-
-
-
Phytochelatin synthetase
-
-
-
-
Synthetase, glutathione
-
-
-
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Glutathione synthetase
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-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
carboxamide formation
-
-
-
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carboxylic acid-amide formation
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-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
-
-, -
SYSTEMATIC NAME
IUBMB Comments
gamma-L-glutamyl-L-cysteine:glycine ligase (ADP-forming)
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CAS REGISTRY NUMBER
COMMENTARY
9023-62-5
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
-
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
-
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
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
-
gene is regulated by stresses via the Atf1-Spc-1-Wis1 signal pathway
-
?
additional information
?
-
no activity with beta-alanine and L-serine by wild-type glutathione synthetase
-
-
?
additional information
?
-
-
no activity with beta-alanine and L-serine by wild-type glutathione synthetase
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
-
gene is regulated by stresses via the Atf1-Spc-1-Wis1 signal pathway
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
acetate
-
anions that favor activity in decreasing order: acetate, HCOO-, Cl-, at 50 mM K+
Cl-
-
anions that favor activity in decreasing order: acetate, HCOO-, Cl-, at 50 mM K+
HCOO-
-
anions that favor activity in decreasing order: acetate, HCOO-, Cl-, at 50 mM K+
K+
K+
-
strict requirement for monovalent cation, maximal activity with K+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Hg2+
-
wild-type cells are not able to grow on mercuric chloride containing substrate, but the recombinant strain is able to survive at 0.01 mM mercuric chloride concentration, the wild-type can grow on 0.01 mM Hg2+ in presence of 20 mM GSH
L-buthionine-(S,R)-sulfoximine
-
induction of enzyme expression at 0.01 mM
menadione
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wild-type cells are not able to grow on menadione containing substrate, but the recombinant strain is able to survive at 0.1 mM menadione, induction of enzyme expression by superoxide generation
additional information
-
no induction of enzyme expression by thioacetamide
-
Cd2+
-
wild-type cells are not able to grow on cadmium chloride containing substrate, but the recombinant strain is able to survive at 1 mM cadmium chloride concentration, the wild-type can grow on 1 mM Cd2+ in presence of 20 mM GSH
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
17.2
-
purified recombinant wild-type enzyme after complete proteoltic cleavage
21.9
-
purified recombinant enzyme mutant with eliminated cleavage site
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
homoglutathione synthetase, EC 6.3.2.23, has evolved from glutathione synthetase by a single gene duplication event
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
additional information
additional information
structure comparisons and homology structure molecular modelling of the GSH2 wild-type and mutant enzymes, overview
additional information
-
structure comparisons and homology structure molecular modelling of the GSH2 wild-type and mutant enzymes, overview
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
56000
x * 56000, about, recombinant His-tagged wild-type and mutant enzymes, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
x * 56000, about, recombinant His-tagged wild-type and mutant enzymes, SDS-PAGE
additional information
additional information
structure comparisons and homology structure molecular modelling of the GSH2 wild-type and mutant enzymes, overview
additional information
-
structure comparisons and homology structure molecular modelling of the GSH2 wild-type and mutant enzymes, overview
additional information
-
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
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
glycoprotein
-
Gal-GlyNAc located at the terminal of the oligosaccharide chain
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
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
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
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
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
recombinant His-tagged wild-type and mutant enzymes from Schizosaccharomyces pombe mutant strain, to homogeneity
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
sequence comparison, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain DH5alpha
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
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expression in Schizosaccharomyces pombe, results in 1.4fold higher glutathione content and 1.9fold ncreased enzyme activity, regulation by the Atf1-Spc-1-Wis1 signal pathway
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wang, C.L.; Oliver, D.J.
Glutathione synthetase: similarities of the protein from Schizosaccharomyces pombe and Arabidopsis thaliana
Biochem. J.
326
563-566
1997
Arabidopsis sp., Schizosaccharomyces pombe
Nakagawa, C.W.; Mutoh, N.; Hayashi, Y.
Glutathione synthetase from the fission yeast. Purification and its unique heteromeric subunit structure
Biochem. Cell Biol.
71
447-453
1993
Schizosaccharomyces pombe
Kim, S.J.; Shin, Y.H.; Kim, K.; Park, E.H.; Sa, J.H.; Lim, C.J.
Regulation of the gene encoding glutathione synthetase from the fission yeast
J. Biochem. Mol. Biol.
36
326-331
2003
Schizosaccharomyces pombe
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
J. Biol. Chem.
278
40152-40161
2003
Schizosaccharomyces pombe
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