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Information on EC 6.3.2.2 - glutamate-cysteine ligase and Organism(s) Saccharomyces cerevisiae and UniProt Accession P32477

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IUBMB Comments
Can use L-aminohexanoate in place of glutamate.
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This record set is specific for:
Saccharomyces cerevisiae
UNIPROT: P32477
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The taxonomic range for the selected organisms is: Saccharomyces cerevisiae
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
gcl, gamma-glutamylcysteine synthetase, glutamate-cysteine ligase, gamma-gcs, glutamate cysteine ligase, gamma-glutamylcysteine ligase, gamma-ecs, glclc, gammagcs, gamma-gc, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
gamma-Glutamylcysteine synthetase
-
Gamma-ECS
-
-
-
-
gamma-Glutamyl-L-cysteine synthetase
-
-
-
-
gamma-Glutamylcysteine synthetase
gamma-Glutamylcysteinyl-synthetase
-
-
-
-
GCS
-
-
-
-
Synthetase, gamma-glutamylcysteine
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxylic acid amide formation
-
-
-
-
carboxamide formation
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
L-glutamate:L-cysteine gamma-ligase (ADP-forming)
Can use L-aminohexanoate in place of glutamate.
CAS REGISTRY NUMBER
COMMENTARY hide
9023-64-7
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
show the reaction diagram
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
show the reaction diagram
additional information
?
-
-
Met4 regulates the GSH1 expression in response to GSH availability, model for genetic regulation and control of GSH biosynthesis
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
show the reaction diagram
rate limiting and first step in glutathione biosynthesis, GSH metabolism, overview
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
show the reaction diagram
additional information
?
-
-
Met4 regulates the GSH1 expression in response to GSH availability, model for genetic regulation and control of GSH biosynthesis
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
buthionine sulfoximine
-
gamma-L-Glu-L-Cys
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glutathione
feedback-regulation. The structure of the GCL-glutathione complex to 2.5 A resolution indicates that the inhibitor occupies both the glutamate- and the presumed cysteine-binding site and disrupts the previously observed Mg2+-coordination in the ATP-binding site
L-buthionine-S-sulfoximine
mechanism-based inhibitor. The crystal structure of the enzyme complex to 2.2 A resolution confirms that L-buthionine-S-sulfoximine is phosphorylated on the sulfoximine nitrogen to generate the inhibitory species and reveals contacts that likely contribute to transition state stabilization
methionine
-
inhibits induction of GSH1 expression, independently of GSH
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
H2O2
induction of in enzyme expression and activity
mcCDC34
-
an ubiquitine conjugated protein, induces gamma-glutamylcysteine synthetase expression only in glutathione synthetase-dficient mutants, not in the wild-type
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.08
ATP
wild-type, pH 8.0
0.17
L-Cys
wild-type, pH 8.0
1.21 - 2.15
L-Glu
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.12
gamma-L-Glu-L-Cys
wild-type, pH 8.0
3.93 - 4.7
L-Glu
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
78300
x * 78300, catalytic subunit
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 78300, catalytic subunit
monomer
-
1 * 78200
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structure of the GCL-glutathione complex to 2.5 A resolution indicates that the inhibitor occupies both the glutamate- and the presumed cysteine-binding site and disrupts the previously observed Mg2+-coordination in the ATP-binding site. The structure of the complex with mechanism-based inhibitor L-buthionine-S-sulfoximine to 2.2 A resolution confirms that L-buthionine-S-sulfoximine is phosphorylated on the sulfoximine nitrogen to generate the inhibitory species and reveals contacts that likely contribute to transition state stabilization
structures of glutamate cysteine ligase in the presence of glutamate and MgCl2, to 2.1 A resolution, and in complex with glutamate, MgCl2, and ADP to 2.7 A resolution. Structures reveal an unusual binding pocket for the alpha-carboxylate of the glutamate substrate and an ATP-independent Mg2+ coordination site, clarifying the Mg2+-dependence of the enzymatic reaction
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C266A
about 2fold increase in both Km and Ki value
C266S
about 2fold increase in both Km and Ki value
additional information
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination and analysis, gene GSH1 maps to chromosome X, expression in and functional complementation of an enzyme-deficient mutant strain, the yAP-1 responsive element in the promotor of gene GSH1 is involved in transcription of the gene in response to exposure to cadmium or hydrogen peroxide
a recombinant plasmid, pGMF, containing a gamma-glutamylcysteine synthetase gene (GSH-I) from Saccharomyces cerevisiae, is constructed with a copper-resistance gene as the selection marker and introduced into Saccharomyces cerevisiae YSF-31. The glutathione content of the recombinant strain is 1.5fold (13.1 mg/g*dry cells) of that in the host strain
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DNA sequence determination and analysis, genetic regulation involving AP-1, overview
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expression of the enzyme at high level in deletion mutant strains of GHS1 and GSH2, the latter encoding the glutathione synthetase, the GSH1 promotor is Met4-inducible and GSH repressible
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Griffith, O.W.; Mulcahy, R.T.
The enzymes of glutathione synthesis: gamma-glutamylcysteine synthetase
Adv. Enzymol. Relat. Areas Mol. Biol.
73
209-267
1999
Ascaris suum, Bos taurus, [Candida] boidinii, Ovis aries, Nicotiana tabacum, no activity in Entamoeba histolytica, Proteus mirabilis, Sus scrofa, Xenopus sp., no activity in Giardia sp., Mus musculus (A0A0H2UNM8), Mus musculus (P97494), Escherichia coli (P0A6W9), Rattus norvegicus (P19468), Rattus norvegicus (P48508), Saccharomyces cerevisiae (P32477), Arabidopsis thaliana (P46309), Homo sapiens (P48506), Homo sapiens (P48507), Homo sapiens, Leishmania tarentolae (P90557), Schizosaccharomyces pombe (Q09768), Trypanosoma brucei (Q26820), Acidithiobacillus ferrooxidans (Q56277)
Manually annotated by BRENDA team
Wild, A.C.; Mulcahy, R.T.
Regulation of gamma-glutamylcysteine synthetase subunit gene expression: insights into transcriptional control of antioxidant defenses
Free Radic. Res.
32
281-301
2000
Saccharomyces cerevisiae, Homo sapiens, Mus musculus, Rattus norvegicus
Manually annotated by BRENDA team
Wheeler, G.L.; Quinn, K.A.; Perrone, G.; Dawes, I.W.; Grant, C.M.
Glutathione regulates the expression of gamma-glutamylcysteine synthetase via the Met4 transcription factor
Mol. Microbiol.
46
545-556
2002
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Fan, X.; He, X.; Guo, X.; Qu, N.; Wang, C.; Zhang, B.
Increasing glutathione formation by functional expression of the gamma-glutamylcysteine synthetase gene in Saccharomyces cerevisiae
Biotechnol. Lett.
26
415-417
2004
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Biterova, E.I.; Barycki, J.J.
Mechanistic details of glutathione biosynthesis revealed by crystal structures of Saccharomyces cerevisiae glutamate cysteine ligase
J. Biol. Chem.
284
32700-32708
2009
Homo sapiens, Saccharomyces cerevisiae (P32477), Saccharomyces cerevisiae
Manually annotated by BRENDA team
Biterova, E.I.; Barycki, J.J.
Structural basis for feedback and pharmacological inhibition of Saccharomyces cerevisiae glutamate cysteine ligase
J. Biol. Chem.
285
14459-14466
2010
Saccharomyces cerevisiae (P32477), Saccharomyces cerevisiae
Manually annotated by BRENDA team