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Information on EC 6.3.2.2 - glutamate-cysteine ligase and Organism(s) Homo sapiens
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6.3.2.2 glutamate-cysteine ligaseIUBMB Comments
Can use L-aminohexanoate in place of glutamate.
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Word Map
- 6.3.2.2
-
ganglion
- retinal
- nerve
- heme
-
plexiform
- fiber
- sulfoximine
- dismutase
-
coherence
-
tomography
-
buthionine
- s-transferase
- oxygenase-1
-
macular
-
erythroid
- quinone
-
neuroprotective
-
2-related
-
amacrine
-
cytoprotective
- gssg
-
nadph:quinone
- glaucoma
-
nf-e2-related
-
acuity
- transpeptidase
-
sd-oct
-
peripapillary
-
intravitreal
-
nrf2-mediated
-
nrf2-dependent
-
dentate
-
spectral-domain
-
nrf2-are
-
factor-erythroid
-
subgranular
-
glutathione-related
-
gsh-dependent
- phytochelatins
-
l-buthionine-s,r-sulfoximine
- tert-butylhydroquinone
-
oxidoreductase-1
-
ech-associated
-
kelch-like
-
nrf2-regulated
-
fovea
- biotechnology
- medicine
-
perimetry
- synthesis
-
gsh-depleting
- agriculture
- drug development
-
etdrs
- pharmacology
-
spectralis
The taxonomic range for the selected organisms is: Homo sapiens
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
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
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Gamma-ECS
-
-
-
-
gamma-GCS
gamma-Glutamyl-L-cysteine synthetase
-
-
-
-
gamma-glutamylcysteine ligase
gamma-Glutamylcysteine synthetase
gamma-Glutamylcysteinyl-synthetase
-
-
-
-
GCL
GCLC
GCS
glutamate cysteine ligase
Synthetase, gamma-glutamylcysteine
-
-
-
-
gamma-Glutamylcysteine synthetase
-
-
-
-
GCL
-
-
GCS
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine
Cys553 is not the active site residue
-
ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine
enzyme-bound reaction intermediate is a gamma-glutamyl-phosphate, active site cysteine, mechanism
ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine
substrate binding mechanism and catalytic site structure determination and analysis, Arg127 and conserved Cys249 are involved
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
carboxamide formation
-
-
-
-
carboxylic acid amide formation
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
-
-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
L-glutamate:L-cysteine gamma-ligase (ADP-forming)
Can use L-aminohexanoate in place of glutamate.
CAS REGISTRY NUMBER
COMMENTARY
9023-64-7
-
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 + D-Glu + L-2-aminobutyrate
ADP + phosphate + gamma-D-Glu-L-alpha-aminobutyrate
-
-
-
?
ATP + L-Glu + gamma-aminobutyrate
ADP + phosphate + L-Glu-gamma-aminobutyrate
-
-
-
ir
ATP + L-Glu + L-2-aminobutanoate
ADP + phosphate + gamma-L-Glu-2-aminobutanoate
-
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + L-glutamyl-L-cysteine
-
-
-
-
?
glutamate + ATP + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
assay at pH 8.2
-
-
?
ATP + L-Glu + L-2-aminobutyrate
ADP + phosphate + L-Glu-2-aminobutyrate
-
-
-
?
ATP + L-Glu + L-2-aminobutyrate
ADP + phosphate + L-Glu-2-aminobutyrate
-
-
-
?
ATP + L-Glu + L-alpha-aminobutyrate
ADP + phosphate + gamma-L-Glu-L-alpha-aminobutyrate
-
-
-
?
ATP + L-Glu + L-alpha-aminobutyrate
ADP + phosphate + gamma-L-Glu-L-alpha-aminobutyrate
-
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
combines glutamate and cysteine through the gamma carboxylmoiety rather than the alpha carboxyl moiety found in protein amide bonds, imparting resistance to proteolytic degradation
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
reaction can be performed by the catalytic subunit alone, but presence of the regulatory subunit in the holoenzyme increases the activity
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
rate-limiting step in glutathione biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate limiting step in glutathione biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate limiting step in GSH biosynthesis, rare hereditary enzyme deficiency is associated with low erythrocyte levels of the enzyme leading to hemolytic anemia
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate-limiting step in glutathione biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate-limiting step in glutathione biosynthesis
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
rate limiting and first step in glutathione biosynthesis, GSH metabolism, overview
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
rate-limiting enzyme in the GSH biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
rate-limiting step in glutathione biosynthesis, regulation mechanism and model
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
regulation and signaling in GSH de novo synthesis pathway
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
rate-limiting enzyme in glutathione synthesis
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
overexpression of gamma-GCS decreases drug-induced oxidative stress and confers drug resistance
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
treatment of human breast cancer cells with 2-deoxy-D-glucose causes metabolic oxidative stress that is accompanied by increases in steady-state levels of glutamate cysteine ligase mRNA, glutamate cysteine ligase activity and glutathione content
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
upregulation of gamma-glutamate-cysteine ligase is part of the long-term adaptation process to iron accumulation in neuronal SH-SY5Y cells
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
evidence of TNF-alpha-mediated LEDGF induction of gamma-glutamylcysteine synthetase heavy subunit and mRNA expression. TNF-alpha-induced intracellular level of reactive oxygen species is critical for the regulation of the multidomain adaptor protein LEDGF, which subsequently influences cellular glutathione content by regulating transcription of gamma-glutamylcysteine synthetase heavy subunit
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
first rate-limiting step in GSH biosynthesis, GCL is a major determinant of cellular GSH levels, pathway overview
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
GCL is the key glutathione-synthesizing enzyme
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
GCL is the rate-limiting enzyme in glutathione biosynthesis, its catalytic subunit GCLC determines this de novo synthesis. Induction of GCLC is a strategy to enhance the antioxidant capability in cells
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
rate-limiting enzyme in glutathione synthesis, a mutation in the catalytic subunit gene 5'-UTR leads to reduced enzyme activity
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
the enzyme acts endogenously as an antioxidant and is involved in glutathione biosynthesis
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
GCL-mediated phosphorylation of L-glutamate creating the activated enzyme-bound gamma-glutamylphosphate intermediate
-
-
?
additional information
?
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH, enzyme overexpression provides resistance to melphalan and other drugs, overview, protection of cancer cells by increased GSH levels
-
?
additional information
?
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH, enzyme overexpression provides resistance to melphalan and other drugs, overview, protection of cancer cells by increased GSH levels
-
?
additional information
?
-
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH, enzyme overexpression provides resistance to melphalan and other drugs, overview, protection of cancer cells by increased GSH levels
-
?
additional information
?
-
-
overexpression of cytochrome P450 2E1 in human hepatocarcinoma cell line HepG2 increases the intracellular H2O2 level by 40-50% and therefore results in a 2fold increase in enzyme expression
-
?
additional information
?
-
-
the genotype of GLCLC is associated with drug sensitivity or resistance, respectively
-
?
additional information
?
-
-
GCL activity is not associated with susceptibility to chronic obstructive pulmonary disease patients or disease severity, overview
-
-
?
additional information
?
-
-
GCLC polymorphisms are associated with lower lung function levels causing lung disease, especially in association with oxidative stress due to smoking
-
-
?
additional information
?
-
-
insulin stimulation of GCL catalytic subunit expression increases endothelial GSH during oxidative stress, overview. Functional importance of insulin in Nrf2-dependent transcriptional upregulation of GCLC in GSH recovery during oxidative challenge, role for Nrf2 involvement in both constitutive and inducible endothelial GCLc expression and GSH synthesis, while PI3K/Akt/mTOR signaling appears to participate only in insulin-inducible GSH synthesis. Low glucose enhances the insulin-mediated increase in GCLc expression
-
-
?
additional information
?
-
-
post-translational regulation of GCL, overview. GCLC and GCLM polymorphisms increase disease susceptibility in humans, overview
-
-
?
additional information
?
-
-
the cis-element signaling of Nrf2/EpRE is involved in resveratrol-mediated induction of GCL genes
-
-
?
additional information
?
-
-
the modifier subunit GCLM is not correlated with methamphetamine-use disorder or schizophrenia in the Japanese population, overview
-
-
?
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 + L-glutamate + L-cysteine
ADP + phosphate + L-glutamyl-L-cysteine
-
-
-
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
rate-limiting step in glutathione biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate limiting step in glutathione biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate limiting step in GSH biosynthesis, rare hereditary enzyme deficiency is associated with low erythrocyte levels of the enzyme leading to hemolytic anemia
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate-limiting step in glutathione biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate-limiting step in glutathione biosynthesis
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
rate limiting and first step in glutathione biosynthesis, GSH metabolism, overview
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
rate-limiting enzyme in the GSH biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
rate-limiting step in glutathione biosynthesis, regulation mechanism and model
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
regulation and signaling in GSH de novo synthesis pathway
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
rate-limiting enzyme in glutathione synthesis
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
overexpression of gamma-GCS decreases drug-induced oxidative stress and confers drug resistance
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
treatment of human breast cancer cells with 2-deoxy-D-glucose causes metabolic oxidative stress that is accompanied by increases in steady-state levels of glutamate cysteine ligase mRNA, glutamate cysteine ligase activity and glutathione content
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
upregulation of gamma-glutamate-cysteine ligase is part of the long-term adaptation process to iron accumulation in neuronal SH-SY5Y cells
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
first rate-limiting step in GSH biosynthesis, GCL is a major determinant of cellular GSH levels, pathway overview
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
GCL is the key glutathione-synthesizing enzyme
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
GCL is the rate-limiting enzyme in glutathione biosynthesis, its catalytic subunit GCLC determines this de novo synthesis. Induction of GCLC is a strategy to enhance the antioxidant capability in cells
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
rate-limiting enzyme in glutathione synthesis, a mutation in the catalytic subunit gene 5'-UTR leads to reduced enzyme activity
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
the enzyme acts endogenously as an antioxidant and is involved in glutathione biosynthesis
-
-
?
additional information
?
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH, enzyme overexpression provides resistance to melphalan and other drugs, overview, protection of cancer cells by increased GSH levels
-
?
additional information
?
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH, enzyme overexpression provides resistance to melphalan and other drugs, overview, protection of cancer cells by increased GSH levels
-
?
additional information
?
-
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH, enzyme overexpression provides resistance to melphalan and other drugs, overview, protection of cancer cells by increased GSH levels
-
?
additional information
?
-
-
overexpression of cytochrome P450 2E1 in human hepatocarcinoma cell line HepG2 increases the intracellular H2O2 level by 40-50% and therefore results in a 2fold increase in enzyme expression
-
?
additional information
?
-
-
the genotype of GLCLC is associated with drug sensitivity or resistance, respectively
-
?
additional information
?
-
-
GCL activity is not associated with susceptibility to chronic obstructive pulmonary disease patients or disease severity, overview
-
-
?
additional information
?
-
-
GCLC polymorphisms are associated with lower lung function levels causing lung disease, especially in association with oxidative stress due to smoking
-
-
?
additional information
?
-
-
insulin stimulation of GCL catalytic subunit expression increases endothelial GSH during oxidative stress, overview. Functional importance of insulin in Nrf2-dependent transcriptional upregulation of GCLC in GSH recovery during oxidative challenge, role for Nrf2 involvement in both constitutive and inducible endothelial GCLc expression and GSH synthesis, while PI3K/Akt/mTOR signaling appears to participate only in insulin-inducible GSH synthesis. Low glucose enhances the insulin-mediated increase in GCLc expression
-
-
?
additional information
?
-
-
post-translational regulation of GCL, overview. GCLC and GCLM polymorphisms increase disease susceptibility in humans, overview
-
-
?
additional information
?
-
-
the cis-element signaling of Nrf2/EpRE is involved in resveratrol-mediated induction of GCL genes
-
-
?
additional information
?
-
-
the modifier subunit GCLM is not correlated with methamphetamine-use disorder or schizophrenia in the Japanese population, overview
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ATP
-
-
649413, 649655, 649660, 651391, 651392, 651393, 651996, 653993, 690385, 690440, 690630, 692442, 692444, 694609, 726668
ATP
the gamma-phosphate is located close to the glutamate binding site by the glycine-rich P-loop, built by the motif M(A/G)FGMGXXCLQ, to facilitate the formation of the enzyme-bound reaction intermediate gamma-glutamyl-phosphate
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
glutathione
-
feedback inhibition, subunit GCLM increases the Ki for GSH-mediated feedback inhibition of GCL, competitive to glutamate
NF-kappaB
-
inhibits induction of enzyme expression by other substances, e.g. buthionine sulfoximine or tert-butylhydroquinone
-
buthionine sulfoximine
-
GCL mediates the phosphorylation of buthionine sulfoximine, which is required for its tight and irreversible binding to the active site of GCL
cysteamine
-
inactivation of wild-type enzyme and mutant C553G after 90 min at 4°C, 0.2 mM cysteamine and 2 mM ATP
dithiothreitol
-
inhibition of wild-type holoenzyme and C553G mutant holoenzyme, the latter is more sensitive
GSH
-
noncompetitive to L-glutamate, inhibition is not dependent on reduction of disulfide bonds between the 2 subunits in the holoenzyme
additional information
-
the genotype of GLCLC is associated with drug sensitivity or resistance, respectively, sensitivity of different genotypes to diverse drugs, overview
-
additional information
-
oxidative stress dramatically affects GCL holoenzyme formation and activity
-
additional information
-
decrease of enzyme activity in hypoxia: 20% after 6 h, 17% after 12 h, 23% after 24 h, hypoxia-induced decrease in enzyme activity may be prevented by MAPK inhibition and catalase
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1-(4-amino-2-methyl-5-pyridimidyl)-methyl-3-(2-chloroethyl)-3-nitrosurea
-
induces expression of heavy subunit
2,3-dimethoxy-1,4-naphthoquinone
-
induces expression of heavy and light subunit
apigenin
-
nearly 2fold induction of the heavy subunit gene promotor and heavy subunit expression
caffeic acid
-
treatment of the cells with 100 and 500 microg/ml of caffeic acid increases gamma-GCS activities by 1.4- and 1.8fold compared to the control group, respectively. At the same doses of caffeic acid, the treated cells show increased levels of glutathione by 1.7- and 2.7fold compared to the control, respectively
kaempferol
-
2fold induction of the heavy subunit gene promotor and heavy subunit expression
onion extract
-
containing flavonoids, which increase the expression of both subunits of the enzyme in COS-1 cells
-
oxidative stress
-
activation of GCL occurrs within min of treatment and without any change in GCL protein levels, and coincides with an increase in the proportion of GCL catalytic subunit in the holoenzyme form. Likewise, GCL modifier subunit shifts from the monomeric form to holoenzyme and higher molecular weight species. Neither GCL activation, nor the formation of holoenzyme, requires a covalent intermolecular disulfide bridge between GCL catalytic subunit and GCL modifier subunit. In immunoprecipitation studies, a neutralizing epitope associated with enzymatic activity is protected following cellular oxidative stress. Thus, the N-terminal portion of GCL catalytic subunit may undergo a change that stabilizes the GCL holoenzyme. Results suggest a dynamic equilibrium between low- and high-activity forms of GCL, which is altered by transient oxidative stress
-
pyrrolidine dithiocarbamate
-
time-, dose-, and Cu2+-dependent induction and increase in expression levels of the 2 subunits of the enzyme in HepG2 cells, mechanism, can be partially blocked by N-acetylcysteine and by copper chelator bathocuproine disulfonic acid
quercetin
-
3fold induction of the heavy subunit gene promotor and heavy subunit expression, best at 0.05 mM, induction even of a distal part of the promotor sequence containing only 2 antioxidant-response/electrophile-response elements, i.e. ARE/EpRE
4-hydroxy-2-nonenal
-
i.e. 4-HNE, inductor of enzyme expression, signals through the JNK pathway, most effective at 0.02 mM, time-dependence is different for GCLC and GCLM, overview, also activates the transcription factor complex AP-1 which itself activates expression of both enzyme subunits
4-hydroxy-2-nonenal
-
4-hydroxy-2-nonenal causes a rapid dose- and time-dependent increase in A549 cell GCL activity. Maximal activation of GCL occurs at 30 min in response to 50 microM 4-hydroxy-2-nonenal
AP-1
-
transcription factor complex including Jun family members, drives expression of both enzyme subunit encoding genes, AP-1 complex is activated by 4-hydroxy-2-nonenal, enzyme induction can be blocked by membrane-permeable peptide-based JNK inhibitor JNKi, but not by inhibitors SB202190 or PD98059
-
H2O2
-
40-50% increase in intracellular concentration induces enzyme expression 2fold to upregulate GSH production, GSH is required for detoxification, increase is reversible or preventable by peroxide-eliminating substances
H2O2
induction of enzyme expression, increase in activity in V79 cells independent on transcription level
additional information
-
multiple cis- and trans-elements have up-regulating effect on expression of the heavy catalytic subunit and of the regulatory light subunit, ionization radiation induces expression of heavy subunit, overview, effect of miscellaneous treatments on enzyme mRNA expression, overview
-
additional information
-
no increase in gene promotor activity by myricetin and sugar conjugates of quercetin, quercetin-3-glucoside and quercetin-3-rhamnoglucoside
-
additional information
-
no induction of enzyme production in HepG2 cells by overexpression of human cytochrome 3A4
-
additional information
-
extracts of Ginkgo biloba induces GCL catalytic subunit, GCLC, in HepG2 and Hep1c1c7 cell lines
-
additional information
-
resveratrol and 4-hydroxy-2-nonenal both increases GSH and the mRNA contents of both the catalytic GCLC and modulatory GCLM subunit of GCL, both agents show synergictic effects when applied together, overview. The cis-element Nrf2/EpRE signalling is involved in resveratrol-mediated induction of GCL genes
-
additional information
-
subunit GCLM increases the Vmax and Kcat of subunit GCLC, and decreases the Km for glutamate and ATP
-
additional information
-
the enzyme is induced by 4-hydroxy-2-nonenal through the c-Jun N-terminal kinase pathway, the regulation involves SHP-1, the protein-tyrosine phosphatase SH" domain containing phosphatase-1 in HBE1 cells, mechanism, overview
-
additional information
-
under low glucose levels insulin induces an approximate 2fold increase in expression of gamma-glutamylcysteine ligase catalytic subunit GCLc mRNA and protein, which does not lead to increased GSH levels in the cell
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.014
transformed COS cells expressing both the recombinant subunits at equal amounts
0.038
transformed COS cells expressing the recombinant catalytic subunit
0.12
-
purified recombinant mutant C249G catalytic subunit and mutant C295G catalytic subunit
0.17
-
purified recombinant mutant C52G catalytic subunit and mutant C248G catalytic subunit
0.44
-
purified mutant R127C enzyme, substrates L-glutamate and L-alpha-aminobutyrate
1.15
25
6.17
8.86
-
purified wild-type enzyme, substrates L-glutamate and L-alpha-aminobutyrate
additional information
additional information
-
GSH and GSSG levels in cells after treatment with pyrrolidine dithiocarbamate
additional information
-
the activity of both the recombinant holoenzyme and catalytic subunit increased by 40% after removal of the His-tag
additional information
kinetic measurement, based on an HPLCESI-MS technique: L-2-aminobutyrate is used instead of cysteine as triggering substrate with saturating concentrations of glutamate and ATP, and the gamma-glutamylaminobutyrate formed is measured at m /z = 233 at regular time intervals. The reaction rate is maximum because ATP is held constant by enzymatic recycling of ADP by pyruvate kinase and phosphoenolpyruvate
additional information
-
kinetic measurement, based on an HPLCESI-MS technique: L-2-aminobutyrate is used instead of cysteine as triggering substrate with saturating concentrations of glutamate and ATP, and the gamma-glutamylaminobutyrate formed is measured at m /z = 233 at regular time intervals. The reaction rate is maximum because ATP is held constant by enzymatic recycling of ADP by pyruvate kinase and phosphoenolpyruvate
additional information
-
correlation between enzyme activity and genotype in black and white individuals, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
671216, 671223, 672904, 674140, 675918, 676099, 692442, 692444, 694609, 703592, 703799, 704577, 704665, 704751, 705264, 705828, 706834, 714265, 715088, 716235, 726668
-
-
activity of the gamma-glutamylcysteine synthetase promoter decreases after indomethacin treatment
-
-
enzyme consists of a catalytic GCLC and a modulatory GCLM subunit encoded by 2 separate gcl genes
-
-
genes GCLC and GCLM encoding catalytic and modifier subunits
UniProt
genes GCLC and GCLM encoding heavy and light chain subunits
UniProt
healthy individuals and chronic obstructive pulmonary disease patients, genes gclC and gclM encoding the two subunits of the enzyme
-
-
Japanese individuals and patients with methamphetamine dependence and induced psychosis, genes gclC and gclM encoding the catalytic and the modifier subunit of the enzyme
-
-
leukemia cell. Cells show increased expression of gamma-glutamylcysteine synthetase. Expression decreases by indomethacin treatment
-
-
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
A2780/100 ovarian carcinoma cell exhibits resistance to DNA crosslinking agents, chlorambucil, cisplatin, melphalan, and ionizing radiation compared to the parental cell line, A2780. Drug-resistant cells have the inherent ability to maintain increased gamma-GCS activity
-
treatment of human breast cancer cells with 2-deoxy-D-glucose causes metabolic oxidative stress that is accompanied by increases in steady-state levels of glutamate cysteine ligase mRNA, glutamate cysteine ligase activity and glutathione content
-
oligophosphopeptides derived from egg yolk phosvitin up-regulate gamma-glutamylcysteine synthetase and antioxidant enzymes against oxidative stress in Caco-2 cells
subunit GCLM, but not subunit GCLC, expression is significantly increased in tumor samples, compared with normal mucosa, both at the mRNA and protein level
-
gamma-glutamylcysteine synthetase mediates the c-Myc-dependent response to antineoplastic agents in melanoma cells
the protein expression levels of the GCL subunits (catalytic subunit, GCLc, and modulatory subunit, GCLm) and GCL activity are both significantly increased in renal cancer tissue
additional information
-
both subunits of gamma-GCS are most prominently expressed in bronchiolar epithelium and are expressed generally weakly in alveolar macrophages. In interstitial lung diseases, gamma-GCS is expressed especially in the alveolar and metaplastic alveolar epithelium but is low in the fibrotic areas. The regulation of gamma-GCS is complex, being differentially modulated by various cytokines and durations of exposure, and in various cell types in human lung. Potential mechanism for the low gamma-GCS levels in fibrotic lung diseases may include down-regulation of catalytically active subunit of gamma-GCS
-
upregulation of gamma-glutamate-cysteine ligase is part of the long-term adaptation process to iron accumulation in neuronal SH-SY5Y cells
additional information
expression patterns of both subunits in the tissues, overview, 2 transcript different in size for both the heavy and light subunit occur in the tissues, some tumors overexpress only the heavy subunit rather than the holoenzyme
additional information
expression patterns of both subunits in the tissues, overview, 2 transcript different in size for both the heavy and light subunit occur in the tissues, some tumors overexpress only the heavy subunit rather than the holoenzyme
additional information
-
expression patterns of both subunits in the tissues, overview, 2 transcript different in size for both the heavy and light subunit occur in the tissues, some tumors overexpress only the heavy subunit rather than the holoenzyme
additional information
expression patterns of both subunits in the tissues, overview, 2 transcripts different in size for both the heavy and light subunit occur in the tissues, some tumors overexpress only the heavy subunit rather than the holoenzyme
additional information
expression patterns of both subunits in the tissues, overview, 2 transcripts different in size for both the heavy and light subunit occur in the tissues, some tumors overexpress only the heavy subunit rather than the holoenzyme
additional information
-
expression patterns of both subunits in the tissues, overview, 2 transcripts different in size for both the heavy and light subunit occur in the tissues, some tumors overexpress only the heavy subunit rather than the holoenzyme
additional information
-
genotyping of 60 tumor cell lines from diverse source tissues for the GLCLC trinuleotide repeat
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
additional information
additional information
-
while GCLC and GCLM are generally considered to be cytosolic proteins there is evidence that they may exhibit altered subcellular localization in certain circumstances
-
additional information
the expression of subunit GCLM is co-localized with the expression of the proliferation marker Ki?67
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
metabolism
glutamate cysteine ligase (GCL) catalyzes the first and rate-limiting step of glutathione, GSH, biosynthesis. The associations between GSH levels and GCL activity with demographic characteristics, clinical manifestations and laboratory parameters in peripheral blood mononuclear cells (PBMCs) are analyzed, overview
physiological function
additional information
malfunction
GSH can be depleted through the specific downregulation of the GCL levels by hammerhead ribozyme
malfunction
in patients with systemic lupus erythematosus (SLE), the levels of enzyme GCL activity and reduced glutathione (GSH) decrease, while thioredoxin (TRX) and oxidized glutathione (GSSG) levels increase when compared with those in the healthy controls. GSH concentrations and GCL activity levels negatively correlate with the SLE disease activity index and erythrocyte sedimentation rate. Patients with SLE and nephritis have lower levels of GSH and GCL activity and higher levels of TRX and GSSG compared with those in SLE patients without nephritis. Insufficient levels of GSH and GCL activity in PBMCs may contribute to the pathogenesis of SLE. A negative association of GSH levels with T-lymphocyte and CD4+ and CD8+ lymphocyte subset apoptosis, and intracellular activated caspase?3 may supports the role of GSH in the alteration of apoptosis of T lymphocytes in the SLE disease state. GSH is involved in the depletion of CD4+ T lymphocytes in patients with SLE
physiological function
-
model to explain adenosine triphosphate depletion during cystinosis. In the absence of cysteine, enzyme gamma-glutamyl cysteine synthetase forms 5-oxoproline, and the 5-oxoproline is converted into glutamate by the ATP-dependant enzyme, 5-oxoprolinase. Thus, in cysteine-limiting conditions, glutamate is cycled back into glutamate via 5-oxoproline at the cost of two ATP molecules without production of glutathione and this is the cause of the decreased levels of glutathione synthesis, as well as the ATP depletion observed in these cells. The model is also compatible with the differences seen in the human patients and the mouse model of cystinosis, where renal failure is not observed
physiological function
-
the high resistance of MYCN-amplified neuroblastoma cells against oxidative damage can be accounted for by their greater expression of both the mRNA and protein of the catalytic subunit of glutamate-cysteine ligase, the rate-limiting step in GSH biosynthesis. MYCN directly binds to an E-box containing GCL catalytic subunit promoter and over-expression of MYCN in MYCN-non-amplified cells stimulates GCL catalytic subunit expression and provides resistance to oxidative damage. Knock-down of MYCN in MYCN-amplified cells decreases GCL catalytic subunit expression and sensitizes them to oxidative damage. GCL catalytic subunit knock-down enhances the vulnerability of MYCN-amplified cells to oxidative damage
physiological function
-
transfection of COV-434 granulosa tumour cell with vectors designed for the constitutive expression of Gcl catalytic subunit, Gcl modifier subunit, or both Gcl catalytic subunit and Gcl modifier subunit. GCL protein and enzymatic activity and total GSH levels are significantly increased in the GCL subunit-transfected cells. GCL-transfected cells are resistant to cell killing by treatment with hydrogen peroxide compared to control cells. In all the GCL subunit-transfected cell lines cell viability declines less than in control 1-8 h after 0.5 mM hydrogen peroxide treatment. In cells irradiated with 0, 1 or 5 Gy of g-rays, there is a dose-dependent increase in reactive oxygen species within 30 min in all cell lines, this effect is significantly attenuated in Gcl-transfected cells. Apoptosis is significantly decreased in irradiated Gclc-transfected cells compared to irradiated control cells
physiological function
-
expression of both glutamate-cysteine ligase catalytic and modifier subunit is mediated by the GCN2/ATF4 stress response pathway. Regulation of modifier subunit GCLM expression may be mediated by changes in the abundance of mRNA stabilizing or destabilizing proteins. Upregulation of GCLM levels in response to low cysteine levels may serve to protect the cell in the face of a future stress requiring GSH as an antioxidant or conjugating/detoxifying agent
physiological function
GSH is synthesized de novo in a two-step process catalyzed by glutamate cysteine ligase (GCL, EC 6.3.2.2), and glutathione synthetase (GS, EC 6.3.2.3). GCL catalyzes the first and rate-limiting step, in which glutamate is ligated with cysteine to form gamma-glutamylcysteine (gamma-GC), which is rapidly linked to glycine to form GSH via the action of glutathione synthetase. Increased expression and enzymatic activity of enzyme GCL is closely associated with renal cell carcinoma (RCC) suggesting an important role for glutathione in the pathogenesis of RCC
physiological function
the enzyme catalyzes the first rate-limiting step in the biosynthesis of glutathione. Glutathione (GSH) is the keystone of the cellular response toward oxidative stress. Elevated GSH content correlates with increased resistance to chemotherapy and radiotherapy of head and neck (HN) tumors. Nuclear localization of glutamate-cysteine ligase is associated with proliferation in head and neck squamous cell carcinoma. The localization of GSH synthesis contributes to the protection against oxidative stress within hotspots of cell proliferation. The expression of glutamate-cysteine ligase (GCL) accounts for the increased GSH availability observed in head and neck squamous cell carcinoma (SCC). No role of the NRF2 and NF?kappaB signaling pathways in GCLM activation
additional information
GCL is a heterodimeric enzyme, consisting of a catalytic subunit, (GCLc) and a modulatory subunit (GCLm) that are encoded by two distinct genes. GCLc constitutes all the enzymatic activity, but catalytic efficiency is increased substantially by covalent interaction with GCLm
additional information
-
GCL is a heterodimeric enzyme, consisting of a catalytic subunit, (GCLc) and a modulatory subunit (GCLm) that are encoded by two distinct genes. GCLc constitutes all the enzymatic activity, but catalytic efficiency is increased substantially by covalent interaction with GCLm
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). GSH1_HUMAN
637
0
72766
Swiss-Prot
other Location (Reliability: 4)
Q14TF0_HUMAN
637
0
72766
TrEMBL
other Location (Reliability: 4)
A0A2R8YEL6_HUMAN
639
0
72932
TrEMBL
other Location (Reliability: 4)
D6RGF8_HUMAN
54
0
6356
TrEMBL
other Location (Reliability: 4)
A0A2R8Y648_HUMAN
361
0
41344
TrEMBL
other Location (Reliability: 3)
E1CEI4_HUMAN
599
0
68630
TrEMBL
other Location (Reliability: 4)
A0A0C4DGB2_HUMAN
252
0
28752
TrEMBL
other Location (Reliability: 4)
B4E2I4_HUMAN
584
0
66464
TrEMBL
other Location (Reliability: 1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
114000
30700
1 * 72800, heavy catalytic subunit, + 1 * 30700, light regulatory subunit, SDS-PAGE
31000
72800
1 * 72800, heavy catalytic subunit, + 1 * 30700, light regulatory subunit, SDS-PAGE
73000
31000
-
1 * 73000, about, catalytic subunit, + 1 * 31000, about, regulatory subunit, SDS-PAGE
73000
-
1 * 73000, about, catalytic subunit, + 1 * 31000, about, regulatory subunit, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
heterodimer
additional information
dimer
1 * 72800, heavy catalytic subunit, + 1 * 30700, light regulatory subunit, SDS-PAGE
dimer
-
1 * 73000, about, catalytic subunit, + 1 * 31000, about, regulatory subunit, SDS-PAGE
dimer
-
heterodimer of a catalytic subunit and a regulatory subunit, encoded by 2 genes
dimer
-
the enzyme consists of a catalytic subunit GCLC and a modifier subunit GCLM
heterodimer
enzyme GCL is a heterodimeric enzyme consisting of a catalytic subunit (GCLc) and a modulatory subunit (GCLm), which are encoded by two genes. The catalytic subunit GCLc performs all the enzymatic activity and its catalytic efficiency is increased by the covalent interaction with the regulatory subunit GCLm
heterodimer
GCL is a heterodimeric enzyme, consisting of a catalytic subunit, (GCLc) and a modulatory subunit (GCLm) that are encoded by two distinct genes. GCLc constitutes all the enzymatic activity, but catalytic efficiency is increased substantially by covalent interaction with GCLm
additional information
-
enzyme consists of 2 subunits, the heavy catalytic one and the light regulatory one
additional information
-
enzyme consists of a catalytic GCLC and a modulatory GCLM subunit
additional information
quarternary structure, the heavy subunit monomer may be essentially nonfunctional under physiological conditions
additional information
quarternary structure, the heavy subunit monomer may be essentially nonfunctional under physiological conditions
additional information
-
quarternary structure, the heavy subunit monomer may be essentially nonfunctional under physiological conditions
additional information
-
reaction can be performed by the catalytic subunit alone, but presence of the regulatory subunit in the holoenzyme increases the activity
additional information
-
reaction can be performed by the catalytic subunit alone, but presence of the regulatory subunit in the holoenzyme increases the activity and the specificity with L-2-aminobutyrate as substrate
additional information
-
the holoenzyme consists of a heavy catalytic and a light modifier subunit, i.e. gamma-GCSH and gamma-GCSL, protein modeling
additional information
-
the holoenzyme consists of a heavy catalytic and a light regulatory subunit, i.e. gamma-GCSh and gamma-GCSl
additional information
-
GCL is a heterodimeric protein composed of catalytic GCLC and modifier GCLM subunits that are expressed from different genes, the catalytic subunit GCLC contains the active site responsible for the ATP-dependent bond formation between the amino group of cysteine and the gamma-carboxyl group of glutamate, the modifier subunit GCLM through direct interaction with GCLC acts to increase the catalytic efficiency of GCLC. GCL subunit protein structures, overview. GCLM is quite sensitive to aggregation in vitro in the absence of GCLC
additional information
-
upon oxidative stress, activation of GCL occurrs within min of treatment and without any change in GCL protein levels, and coincides with an increase in the proportion of GCL catalytic subunit in the holoenzyme form. Likewise, GCL modifier subunit shifts from the monomeric form to holoenzyme and higher molecular weight species. Neither GCL activation, nor the formation of holoenzyme, requires a covalent intermolecular disulfide bridge between GCL catalytic subunit and GCL modifier subunit
additional information
-
the enzyme consists of a catalytic (GCLC) and a modifier (GCLM) subunit
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
lipoprotein
-
myristoylation is responsible for regulation of GCL subunit subcellular localization to membranes and mitochondria, overview
phosphoprotein
-
phosphorylation plays an important role in regulating GCL activity in vivo, phosphorylation of GCLC occurs on serine and threonine residues in vitro and the phosphorylation sites are likely identical for all three kinases protein kinase C, PKC, cAMP-dependent protein kinase, PKA, or Ca2+-calmodulin-dependent protein kinase II, CMKII
additional information
-
post-translational modifications of GCLC, e.g. phosphorylation, myristoylation, caspase-mediated cleavage, have modest effects on GCL activity
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
homology model of the catalytic subunit of human glutamate cysteine ligase. Examination of the model suggests that post-translational modifications of cysteine residues may be involved in the regulation of enzymatic activity
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C248G
-
site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme
C249G
-
site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, reduced activity of the holoenzyme compared to the wild-type enzyme
C295G
-
site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme
C491G
-
site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme
C501G
-
site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme
C52G
-
site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme
C553G
-
site-directed mutagenesis in the catalytic subunit, slightly reduced activity of the catalytic subunit, about 3.5fold reduced activity of the holoenzyme compared to the wild-type enzyme
C605G
-
site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme
H370L
-
clinically relevant mutation in catalytic subunit GCLC. Significantly lower levels of glutathione relative to that of the wild type. Compromised enzymatic activity can largely be rescued by the addition of GCLM
P158L
-
clinically relevant mutation in catalytic subunit GCLC. Significantly lower levels of glutathione relative to that of the wild type, kinetic constants comparable to those of wild-type GCLC
P414L
-
clinically relevant mutation in catalytic subunit GCLC. Significantly lower levels of glutathione relative to that of the wild type, most compromised mutant among those studied. Compromised enzymatic activity can largely be rescued by the addition of GCLM
P462S
-
non-synonymous polymorphism in the gene encoding the catalytic subunit of glutamate-cysteine ligase. The polymorphism is present only in individuals of African descent and encodes an enzyme with significantly decreased in vitro activity when expressed by either a bacterial or mammalian cell expression system. Overexpression of the P462 wild-type GCLC enzyme results in higher intracellular glutathione concentrations than overexpression of the P462S isoform. Apoptotically stimulated mammalian cells overexpressing the P462S enzyme have increased caspase activation and increased DNA laddering compared to cells overexpressing the wild-type enzyme. The P462S polymorphism is in Hardy-Weinberg disequilibrium, with no individuals homozygous for the P462S polymorphism identified
R127C
additional information
R127C
-
naturally occuring mutation in patients with hemolytic anemia resulting from low erythrocyte enzyme levels, fibroblast cells from these patients bearing the mutation show 95% reduced enzyme activity and lowered Km for glutamate and aminobutyrate, as well as an increased Ki for GSH
R127C
-
clinically relevant mutation in catalytic subunit GCLC. Significantly lower levels of glutathione relative to that of the wild type. Compromised enzymatic activity can largely be rescued by the addition of GCLM
additional information
mutational analysis of the 5'-flanking sequence of the heavy subunit, site-directed mutagenesis
additional information
mutational analysis of the 5'-flanking sequence of the heavy subunit, site-directed mutagenesis
additional information
-
mutational analysis of the 5'-flanking sequence of the heavy subunit, site-directed mutagenesis
additional information
-
a GAG-repeat polymorphism in the 5'-UTR of the gene coding for the catalytic subunit, GCLC, is associated with altered GSH levels in vitro and in vivo
additional information
-
GCLC and GCLM polymorphisms increase disease susceptibility in humans, overview
additional information
-
GCLC polymorphisms are associated with lower lung function levels causing lung disease, especially in association with oxidative stress due to smoking
additional information
-
the presence of at least one T allele in the -129 C/T polymorphism of the GCL catalytic subunit gene is independently associated with non-alcoholic steatohepatitis
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
activity of the holoenzyme and of the catalytic subunit is reduced by 20% and 10%, respectively, after 1 cycle of freezing and thawing
-
freezing of the purified recombinant enzyme in solution results in irreversible inactivation
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 25% glycerol, 20 mM imidazole HCl buffer, pH 7.4, 1 mM EDTA, without freezing, stable for at least 1 year
-
4°C, purified recombinant enzyme, 20 mM imidazole HCl buffer, pH 7.4, 1 mM EDTA, stable for at least 7 days
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
from erythrocyte, from malignant astrocytoma cell line, recombinant from Escherichia coli to homogeneity
recombinant His-tagged holoenzyme and individual subunits from Sf9 insect cells
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
catalytic subunit DNA sequence determination and analysis, expression as His-tagged wild-type enzyme and mutant R127C in Rosetta cells, functional expression of wild-type and mutant enzyme in enzyme-deficient cells
-
coexpression of the catalytic and the regulatory subunit from 2 different plasmids in Escherichia coli BL21(DE3), intracellular assembly of the holoenzyme
-
coexpression of the His-tagged catalytic and regulatory subunits in Spodoptera frugiperda Sf9 cells via baculovirus infection, formation of the holoenzyme in the cells
-
DNA sequence determination and analysis, 2 genes encode the 2 subunits, chromosomal mapping to 6p12 and 1p21, constitutive expression, expression of the heavy subunit alone or in combination with the light subunit in mammalian cells reveals that the regulatory subunit improves the activity, genetic regulation involving AP-1, overview, expression of several constructs in HepG2 cells, signaling for enzyme expression
-
DNA sequence determination and analysis, heterodimer of a catalytic subunit and a regulatory subunit, encoded by 2 genes: GLCLC for the catalytic subunit, and GLCLR for the regulatory subunit, GLCLC polymorphism and existence of 5 alleles as defined by the trinucleotide repeat, which exhibits a range of 4 to 10 uninterrupted repeat, genotyping for the repeat of 60 tumor cancer cell lines, overview
-
DNA sequence determination and analysis, mapping to chromosome 6p12, heavy and light subunits, overexpression in Escherichia coli, individual or coexpression of the 2 subunits in COS cells, expression patterns, expression of several deletion mutants created fom the 5'-flanking region of the gene in human hepatoblastoma HepG2 cells, overexpression in human leukemia HL-60 cells
DNA sequence determination and analysis, mapping to chromosome 6p12, heavy and light subunits, overexpression in Escherichia coli, individual or coexpression of the 2 subunits in COS cells, expression patterns, expression of several deletion mutants created fom the 5'-flanking region of the gene in humen hepatoblastoma HepG2 cells, overexpression in human leukemia HL-60 cells
expression of a construct consisting of 3.8 kb of the enzyme's heavy subunit gene promotor in front of luciferase as well as a fragment of the 5'-flanking sequence, transient expression in COS-1 cells
-
expression of the His-tagged wild-type enzyme and mutants in Spodoptera frugiperda Sf9 cells via baculovirus infection
-
generation of C57Bl/6 mice that conditionally overexpress glutamate-cysteine ligase
-
genes gclC and gclM, genotyping in healthy individuals and chronic obstructive pulmonary disease patients
-
genes gclC and gclM, genotyping in healthy individuals and in schizophrenia patients, overview
-
genes gclc and gclm, genotyping, analysis of correlation between genotype and smoking effects, overview
-
genes GCLC and GCLM, quantitative RT-PCR expression analysis in biopsy samples from head and neck squamous cell carcinoma tissues and adjacent normal tissues
transfection of COV-434 granulosa tumour cell with vectors designed for the constitutive expression of Gcl catalytic subunit, Gcl modifier subunit, or both Gcl catalytic subunit and Gcl modifier subunit
-
transfecttion of embryonic fibroblast from GCLC null mice and expression in Saccharomyces cerevisiae
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
both the glutamate-cysteine ligase catalytic (GCLC) and modifier (GCLM) subunit mRNA levels are upregulated in response to a lack of cysteine or other essential amino acids, independent of GSH levels
-
catalytic subunit GCLC protein levels do not increase, whereas regulatory subunit GCLM protein levels increase in the cells cultured in cysteine-deficient medium
-
indomethacin inhibits the gamma-glutamylcysteine synthetase promoter activity. Co-treatment by indomethacin and doxorubicin increases the cytotoxicitiy of doxorubicin by decreasing the intracellular contents of glutathione and its conjugates with decreasing expression of gamma-glutamylcysteine synthetase
-
oncogen MYCN directly binds to an E-box containing GCL catalytic subunit promoter and over-expression of MYCN in MYCN-non-amplified cells stimulates GCL catalytic subunit expression and provides resistance to oxidative damage. Knock-down of MYCN in MYCN-amplified cells decreases GCL catalytic subunit expression and sensitizes them to oxidative damage
-
presence of an ethanol-responsive element in the human GCL catalytic subunit promoter, it spannes bases 1432 to 832 in hepatocytes and HepG2 cells transfected with cytochrome P450 2E1. The region lacks an ARE but has a putative nuclear factor-kappaB element
-
specific downregulation of the GCL levels by hammerhead ribozyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
pharmacology
-
co-treatment by indomethacin and doxorubicin increases the cytotoxicitiy of doxorubicin by decreasing the intracellular contents of glutathione and its conjugates with decreasing expression of gamma-glutamylcysteine synthetase. Indomethacin inhibits the gamma-glutamylcysteine synthetase promoter activity.
medicine
enzyme inhibitor L-buthionine-S-sulfoximine is used to modulate GSH levels in cancer patients, overview
medicine
-
gamma-GCS gene is the downstream target of c-Myc oncoprotein, driving the response to ROS-inducing drugs. gamma-GCS impairment might specifically sensitize high c-Myc tumor cells to chemotherapy
medicine
-
model to explain adenosine triphosphate depletion during cystinosis. In the absence of cysteine, enzyme gamma-glutamyl cysteine synthetase forms 5-oxoproline, and the 5-oxoproline is converted into glutamate by the ATP-dependant enzyme, 5-oxoprolinase. Thus, in cysteine-limiting conditions, glutamate is cycled back into glutamate via 5-oxoproline at the cost of two ATP molecules without production of glutathione and this is the cause of the decreased levels of glutathione synthesis, as well as the ATP depletion observed in these cells. The model is also compatible with the differences seen in the human patients and the mouse model of cystinosis, where renal failure is not observed
medicine
-
the presence of at least one T allele in the -129 C/T polymorphism of the GCL catalytic subunit gene is independently associated with non-alcoholic steatohepatitis
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Sriram, R.; Ali-Osman, F.
Purification and biochemical characterization of gamma-glutamylcysteine synthetase from a human malignant astrocytoma cell line
Biochem. Mol. Biol. Int.
30
1053-1060
1993
Homo sapiens
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)
Tu, Z.; Anders, M.W.
Expression and characterization of human glutamate-cysteine ligase
Arch. Biochem. Biophys.
354
247-254
1998
Homo sapiens
Tu, Z.; Anders, M.W.
Identification of an important cysteine residue in human glutamate-cysteine ligase catalytic subunit by site-directed mutagenesis
Biochem. J.
336
675-680
1998
Homo sapiens
-
Wild, A.C.; Mulcahy, R.T.
Pyrrolidine dithiocarbamate up-regulates the expression of the genes encoding the catalytic and regulatory subunits of gamma-glutamylcysteine synthetase and increases intracellular glutathione levels
Biochem. J.
338
659-665
1999
Homo sapiens
-
Hamilton, D.; Wu, J.H.; Alaoui-Jamali, M.; Batist, G.
A novel missense mutation in the gamma-glutamylcysteine synthetase catalytic subunit gene causes both decreased enzymatic activity and glutathione production
Blood
102
725-730
2003
Homo sapiens
Myhrstad, M.C.; Carlsen, H.; Nordstrom, O.; Blomhoff, R.; Moskaug, J.O.
Flavonoids increase the intracellular glutathione level by transactivation of the gamma-glutamylcysteine synthetase catalytical subunit promoter
Free Radic. Biol. Med.
32
386-393
2002
Homo sapiens
Dickinson, D.A.; Iles, K.E.; Watanabe, N.; Iwamoto, T.; Zhang, H.; Krzywanski, D.M.; Forman, H.J.
4-Hydroxynonenal induces glutamate cysteine ligase through JNK in HBE1 cells
Free Radic. Biol. Med.
33
974-987
2002
Homo sapiens
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
Mari, M.; Cederbaum, A.I.
CYP2E1 overexpression in HepG2 cells induces glutathione synthesis by transcriptional activation of gamma-glutamylcysteine synthetase
J. Biol. Chem.
275
15563-15571
2000
Homo sapiens
Misra, I.; Griffith, O.W.
Expression and purification human g-glutamylcysteine synthetase
Protein Expr. Purif.
13
268-276
1998
Homo sapiens
Walsh, A.C.; Feulner, J.A.; Reilly, A.
Evidence for functionally significant polymorphism of human glutamate cysteine ligase catalytic subunit: association with glutathione levels and drug resistance in the National Cancer Institute tumor cell line panel
Toxicol. Sci.
61
218-223
2001
Homo sapiens
Tiitto, L.H.; Peltoniemi, M.J.; Kaarteenaho-Wiik, R.L.; Soini, Y.M.; Paakko, P.K.; Sormunen, R.T.; Kinnula, V.L.
Cell-specific regulation of gamma-glutamylcysteine synthetase in human interstitial lung diseases
Hum. Pathol.
35
832-839
2004
Homo sapiens
Chik, K.; Flourie, F.; Arab, K.; Steghens, J.P.
Kinetic measurement by LC/MS of gamma-glutamylcysteine ligase activity
J. Chromatogr. B
827
32-38
2005
Rattus norvegicus (P19468), Homo sapiens (P48506), Homo sapiens
Takamura, Y.; Fatma, N.; Kubo, E.; Singh, D.P.
Regulation of heavy subunit chain of gamma-glutamylcysteine synthetase by tumor necrosis factor-alpha in lens epithelial cells: role of LEDGF/p75
Am. J. Physiol.
290
C554-C566
2006
Homo sapiens
Aguirre, P.; Valdes, P.; Aracena-Parks, P.; Tapia, V.; Nunez, M.T.
Upregulation of gamma-glutamate-cysteine ligase as part of the long-term adaptation process to iron accumulation in neuronal SH-SY5Y cells
Am. J. Physiol.
292
C2197-C2203
2007
Homo sapiens
Andringa, K.K.; Coleman, M.C.; Aykin-Burns, N.; Hitchler, M.J.; Walsh, S.A.; Domann, F.E.; Spitz, D.R.
Inhibition of glutamate cysteine ligase activity sensitizes human breast cancer cells to the toxicity of 2-deoxy-D-glucose
Cancer Res.
66
1605-1610
2006
Homo sapiens
Katayama, S.; Ishikawa, S.; Fan, M.Z.; Mine, Y.
Oligophosphopeptides derived from egg yolk phosvitin up-regulate gamma-glutamylcysteine synthetase and antioxidant enzymes against oxidative stress in Caco-2 cells
J. Agric. Food Chem.
55
2829-2835
2007
Homo sapiens
Das, G.C.; Bacsi, A.; Shrivastav, M.; Hazra, T.K.; Boldogh, I.
Enhanced gamma-glutamylcysteine synthetase activity decreases drug-induced oxidative stress levels and cytotoxicity
Mol. Carcinog.
45
635-647
2006
Homo sapiens
Benassi, B.; Zupi, G.; Biroccio, A.
gamma-Glutamylcysteine synthetase mediates the c-Myc-dependent response to antineoplastic agents in melanoma cells
Mol. Pharmacol.
72
1015-1023
2007
Homo sapiens
Rinna, A.; Forman, H.J.
SHP-1 inhibition by 4-hydroxynonenal activates Jun N-terminal kinase and glutamate cysteine ligase
Am. J. Respir. Cell Mol. Biol.
39
97-104
2008
Homo sapiens
Siedlinski, M.; Postma, D.S.; van Diemen, C.C.; Blokstra, A.; Smit, H.A.; Boezen, H.M.
Lung function loss, smoking, vitamin C intake, and polymorphisms of the glutamate-cysteine ligase genes
Am. J. Respir. Crit. Care Med.
178
13-19
2008
Homo sapiens
Kishi, T.; Ikeda, M.; Kitajima, T.; Yamanouchi, Y.; Kinoshita, Y.; Kawashima, K.; Inada, T.; Harano, M.; Komiyama, T.; Hori, T.; Yamada, M.; Iyo, M.; Sora, I.; Sekine, Y.; Ozaki, N.; Ujike, H.; Iwata, N.
Glutamate cysteine ligase modifier (GCLM) subunit gene is not associated with methamphetamine-use disorder or schizophrenia in the Japanese population
Ann. N. Y. Acad. Sci.
1139
63-69
2008
Homo sapiens
Zhang, H.; Shih, A.; Rinna, A.; Forman, H.J.
Resveratrol and 4-hydroxynonenal act in concert to increase glutamate cysteine ligase expression and glutathione in human bronchial epithelial cells
Arch. Biochem. Biophys.
481
110-115
2008
Homo sapiens
Chappell, S.; Daly, L.; Morgan, K.; Guetta-Baranes, T.; Roca, J.; Rabinovich, R.; Lotya, J.; Millar, A.B.; Donnelly, S.C.; Keatings, V.; MacNee, W.; Stolk, J.; Hiemstra, P.S.; Miniati, M.; Monti, S.; OConnor, C.M.; Kalsheker, N.
Genetic variants of microsomal epoxide hydrolase and glutamate-cysteine ligase in COPD
Eur. Respir. J.
32
931-937
2008
Homo sapiens
Langston, W.; Circu, M.L.; Aw, T.Y.
Insulin stimulation of gamma-glutamylcysteine ligase catalytic subunit expression increases endothelial GSH during oxidative stress: Influence of low glucose
Free Radic. Biol. Med.
45
1591-1599
2008
Homo sapiens
Nichenametla, S.N.; Ellison, I.; Calcagnotto, A.; Lazarus, P.; Muscat, J.E.; Richie, J.P.
Functional significance of the GAG trinucleotide-repeat polymorphism in the gene for the catalytic subunit of gamma-glutamylcysteine ligase
Free Radic. Biol. Med.
45
645-650
2008
Homo sapiens
Franklin, C.C.; Backos, D.S.; Mohar, I.; White, C.C.; Forman, H.J.; Kavanagh, T.J.
Structure, function, and post-translational regulation of the catalytic and modifier subunits of glutamate cysteine ligase
Mol. Aspects Med.
30
86-98
2008
Arabidopsis thaliana, Drosophila melanogaster, Homo sapiens, Mus musculus, Rattus norvegicus
Liu, X.P.; Goldring, C.E.; Wang, H.Y.; Copple, I.M.; Kitteringham, N.R.; Park, B.K.; Wei, W.
Extract of Ginkgo biloba induces glutamate cysteine ligase catalytic subunit (GCLC)
Phytother. Res.
22
367-371
2008
Homo sapiens
Asano, T.; Tsutsuda-Asano, A.; Fukunaga, Y.
Indomethacin overcomes doxorubicin resistance by decreasing intracellular content of glutathione and its conjugates with decreasing expression of gamma-glutamylcysteine synthetase via promoter activity in doxorubicin-resistant leukemia cells
Cancer Chemother. Pharmacol.
64
715-721
2009
Homo sapiens
Jackson, R.M.; Gupta, C.
Hypoxia and kinase activity regulate lung epithelial cell glutathione
Exp. Lung Res.
36
45-56
2010
Homo sapiens
Park, H.; Nam, M.; Lee, H.; Jun, W.; Hendrich, S.; Lee, K.
Isolation of caffeic acid from Perilla frutescens and its role in enhancing ?-glutamylcysteine synthetase activity and glutathione level
Food Chem.
119
724-730
2010
Homo sapiens
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
Krejsa, C.M.; Franklin, C.C.; White, C.C.; Ledbetter, J.A.; Schieven, G.L.; Kavanagh, T.J.
Rapid activation of glutamate cysteine ligase following oxidative stress
J. Biol. Chem.
285
16116-16124
2010
Homo sapiens
Kumar, A.; Bachhawat, A.K.
A futile cycle, formed between two ATP-dependant gamma-glutamyl cycle enzymes, gamma-glutamyl cysteine synthetase and 5-oxoprolinase: the cause of cellular ATP depletion in nephrotic cystinosis?
J. Biosci.
35
21-25
2010
Homo sapiens
Oliveira, C.P.; Stefano, J.T.; Cavaleiro, A.M.; Zanella Fortes, M.A.; Vieira, S.M.; Rodrigues Lima, V.M.; Santos, T.E.; Santos, V.N.; de Azevedo Salgado, A.L.; Parise, E.R.; Ferreira Alves, V.A.; Carrilho, F.J.; Correa-Giannella, M.L.
Association of polymorphisms of glutamate-cystein ligase and microsomal triglyceride transfer protein genes in non-alcoholic fatty liver disease
J. Gastroenterol. Hepatol.
25
357-361
2010
Homo sapiens
de Tudela, M.V.; Delgado-Esteban, M.; Cuende, J.; Bolanos, J.P.; Almeida, A.
Human neuroblastoma cells with MYCN amplification are selectively resistant to oxidative stress by transcriptionally up-regulating glutamate cysteine ligase
J. Neurochem.
113
819-825
2010
Homo sapiens
Cortes-Wanstreet, M.M.; Giedzinski, E.; Limoli, C.L.; Luderer, U.
Overexpression of glutamate-cysteine ligase protects human COV434 granulosa tumour cells against oxidative and gamma-radiation-induced cell death
Mutagenesis
24
211-224
2009
Homo sapiens
Kimura, T.; Kawasaki, Y.; Okumura, F.; Sone, T.; Natsuki, R.; Isobe, M.
Ethanol-induced expression of glutamate-cysteine ligase catalytic subunit gene is mediated by NF-kappaB
Toxicol. Lett.
185
110-115
2009
Homo sapiens
Willis, M.N.; Liu, Y.; Biterova, E.I.; Simpson, M.A.; Kim, H.; Lee, J.; Barycki, J.J.
Enzymatic defects underlying hereditary glutamate cysteine ligase deficiency are mitigated by association of the catalytic and regulatory subunits
Biochemistry
50
6508-6517
2011
Homo sapiens
Backos, D.S.; Fritz, K.S.; Roede, J.R.; Petersen, D.R.; Franklin, C.C.
Posttranslational modification and regulation of glutamate-cysteine ligase by the alpha,beta-unsaturated aldehyde 4-hydroxy-2-nonenal
Free Radic. Biol. Med.
50
14-26
2011
Homo sapiens, Mus musculus
Le, T.M.; Willis, A.S.; Barr, F.E.; Cunningham, G.R.; Canter, J.A.; Owens, S.E.; Apple, R.K.; Ayodo, G.; Reich, D.; Summar, M.L.
An ethnic-specific polymorphism in the catalytic subunit of glutamate-cysteine ligase impairs the production of glutathione intermediates in vitro
Mol. Genet. Metab.
101
55-61
2010
Homo sapiens
Sikalidis, A.K.; Mazor, K.M.; Lee, J.I.; Roman, H.B.; Hirschberger, L.L.; Stipanuk, M.H.
Upregulation of capacity for glutathione synthesis in response to amino acid deprivation: regulation of glutamate-cysteine ligase subunits
Amino Acids
46
1285-1296
2014
Homo sapiens, Mus musculus, Rattus norvegicus, Rattus norvegicus Sprague-Dawley
Li, M.; Zhang, Z.; Yuan, J.; Zhang, Y.; Jin, X.
Altered glutamate cysteine ligase expression and activity in renal cell carcinoma
Biomed. Rep.
2
831-834
2014
Homo sapiens (P48506 AND P48507), Homo sapiens
Song, W.; Yuan, J.; Zhang, Z.; Li, L.; Hu, L.
Altered glutamate cysteine ligase activity in peripheral blood mononuclear cells from patients with systemic lupus erythematosus
Exp. Ther. Med.
8
195-200
2014
Homo sapiens (P48506 AND P48507), Homo sapiens
Dequanter, D.; van de Velde, M.; Bar, I.; Nuyens, V.; Rousseau, A.; Nagy, N.; Vanhamme, L.; Vanhaeverbeek, M.; Brohee, D.; Delree, P.; Boudjeltia, K.; Lothaire, P.; Uzureau, P.
Nuclear localization of glutamate-cysteine ligase is associated with proliferation in head and neck squamous cell carcinoma
Oncol. Lett.
11
3660-3668
2016
Homo sapiens (P48506 AND P48507)
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