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Information on EC 6.3.2.2 - glutamate-cysteine ligase and Organism(s) Mus musculus and UniProt Accession P97494
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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: Mus musculus
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-Glutamyl-L-cysteine synthetase
-
-
-
-
gamma-Glutamylcysteine synthetase
gamma-Glutamylcysteinyl-synthetase
-
-
-
-
GCL
GCS
Synthetase, gamma-glutamylcysteine
-
-
-
-
gamma-Glutamylcysteine synthetase
-
-
-
-
GCS
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
carboxylic acid amide formation
-
-
-
-
carboxamide 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 + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
-
ir
ATP + L-Glu + L-2-aminobutyrate
ADP + phosphate + L-Glu-2-aminobutyrate
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + 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
the mechanism of modulation of eukaryotic gamma-glutamylcysteine ligase enzymes may include specific binding of ligands such as pyridine dinucleotide phosphates and reversible protein phosphorylation
-
-
?
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
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 step in glutathione biosynthesis, regulation mechanism
-
ir
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
first step in glutathione biosynthesis
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
rate-limiting enzyme in GSH synthesis. Overexpression of the catalytic and modifier subunits of the enzyme leads to enhanced GCL activity and resistance to TNF-induced apoptosis. Maintenance of mitochondrial integrity is a major mechanism of protection against TNF-induced apoptosis in Hepa-1 cells overexpressing the enzyme
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
rate-limiting enzyme in the glutathione biosynthesis pathway
-
-
?
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
-
rate-limiting enzyme 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
-
?
additional information
?
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH
-
?
additional information
?
-
-
ARE-driven gene expression of Gclc via a MEK/Nrf2 pathway can help to protect macrophages from oxidative stress due to hyperhomocysteinemia
-
-
?
additional information
?
-
-
post-translational regulation of GCL, 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 + gamma-L-glutamyl-L-cysteine
the mechanism of modulation of eukaryotic gamma-glutamylcysteine ligase enzymes may include specific binding of ligands such as pyridine dinucleotide phosphates and reversible protein phosphorylation
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + L-glutamyl-L-cysteine
-
-
-
-
?
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
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 step in glutathione biosynthesis, regulation mechanism
-
ir
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
first step in glutathione biosynthesis
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
rate-limiting enzyme in GSH synthesis. Overexpression of the catalytic and modifier subunits of the enzyme leads to enhanced GCL activity and resistance to TNF-induced apoptosis. Maintenance of mitochondrial integrity is a major mechanism of protection against TNF-induced apoptosis in Hepa-1 cells overexpressing the enzyme
-
-
?
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
rate-limiting enzyme in the glutathione biosynthesis pathway
-
-
?
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
-
rate-limiting enzyme in glutathione biosynthesis
-
-
?
additional information
?
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH
-
?
additional information
?
-
GSH synthesis is controlled by the amount of enzyme, L-cysteine and by feedback inhibition exerted by GSH
-
?
additional information
?
-
-
ARE-driven gene expression of Gclc via a MEK/Nrf2 pathway can help to protect macrophages from oxidative stress due to hyperhomocysteinemia
-
-
?
additional information
?
-
-
post-translational regulation of GCL, overview
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
As3+
-
As3+ coordinately upregulates GCL catalytic subunit and GCL modifier subunit mRNA levels resulting in increased GCL subunit protein expression, holoenzyme formation, and activity. As3+ increases the rate of transcription of both the GCL catalytic subunit and GCL modifier subunit genes and induces the posttranscriptional stabilization of GCL modifier subunit mRNA. The antioxidant N-acetylcysteine abolishes As3+-induced GCL catalytic subunit expression and attenuates induction of GCL modifier subunit. As3+ induction of GCL catalytic subunit and GCL modifier subunit is also differentially regulated by the MAPK signaling pathways and occurrs independent of the Nrf1/2 transcription factors
Mg2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
MgATP2-
although the enzyme preparation shows a strict requirement for MgATP2- for gamma-glutamylcysteine synthesis, preincubation of the homogenates under phosphorylating conditions with MgATP2- also causes a maximal inhibition of 94%, 77%, 85%, 87%, 83% and 95% in cerebellum, hippocampus, brainstem, striatum, cortex and heart
4-hydroxy-2-nonenal
-
treatment with 4-hydroxy-2-nonenal results in the dose-dependent adduction of both monomeric GCLC and GCLM. 4-Hydroxy-2-nonenal-mediated adduction of monomeric GCLC results in a dose-dependent increase in GCLC enzymatic activity. Treatment of GCL holoenzyme causes a dose-dependent decrease in GCL activity. 4-Hydroxy-2-nonenal-mediated inhibition of GCL holoenzyme activity is associated with a reduction in the levels of heterodimeric GCL holoenzyme complex due to increase in high molecular weight complexes. 4-Hydroxy-2-nonenal modification simultaneously activates monomeric GCLC activity and prevents its ability to heterodimerize with GCLM and form functional GCL holoenzyme
acetaminophen
-
treatment promotes the loss of glutamate cysteine ligase in liver. Activation of glycogen synthase kinase 3beta is a key mediator of the initial phase of acetaminophen-induced liver injury through modulating GCL and Mcl-1 degradation, as well as JNK activation in liver. The silencing of glycogen synthase kinase 3beta decreases the loss of hepatic GCL, and promotes greater GSH recovery in liver following acetaminophen treatment
L-cysteine
-
varying glutamic acid concentrations from 5 to 80 mM do not affect GCL activities markedly, whereas cysteine concentrations from 2.5 to 40 mM influence GCL activities substantially in a tissue-dependent manner, about 20 mM L-Cys is optimal in the different tissue, overview. Low doses activate high doses inhibits the enzyme
buthionine sulfoximine
-
GCL mediates the phosphorylation of buthionine sulfoximine, which is required for its tight and irreversible binding to the active site of GCL
buthionine sulfoximine
P97494; O09172
an irreversible GCL inhibitor, abolishes the beta-carotene-induced GSH increase without affecting the beta-carotene-induced GCL protein expression
glutathione
-
feedback inhibition, subunit GCLM increases the Ki for GSH-mediated feedback inhibition of GCL, competitive to glutamate
glutathione
P97494; O09172
compared with the holoenzyme, the catalytic GCLc monomer shows lower enzymatic activity but higher sensitivity to feedback inhibition by GSH
additional information
no inhibition by alpha-ethyl-methionine sulfoximine
-
additional information
no inhibition by alpha-ethyl-methionine sulfoximine
-
additional information
-
oxidative stress dramatically affects GCL holoenzyme formation and activity
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
L-cysteine
-
varying glutamic acid concentrations from 5 to 80 mM do not affect GCL activities markedly, whereas cysteine concentrations from 2.5 to 40 mM influence GCL activities substantially in a tissue-dependent manner, about 20 mM L-Cys is optimal in the different tissue, overview. Low doses activate high doses inhibits the enzyme
additional information
-
ionization radiation induces expression of heavy subunit, effect of miscellaneous treatments on enzyme mRNA expression, overview
-
additional information
-
induction of Gclc by homocysteine, ARE4 plays a direct role in mediating the induction, Nrf2 signalling is critical in homocysteine-induced activation of ARE4, overview
-
additional information
-
subunit GCLM increases the Vmax and Kcat of subunit GCLC, and decreases the Km for glutamate and ATP
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
acoustic overstimulation facilitates the expression of glutamate-cysteine ligase catalytic subunit probably through enhanced DNA binding of activator protein-1 and/or NF-kappaB in the murine cochlea
-
catalytic subunit GCLc and modifier subunit GCLm are differently regulated during development in the placenta and the yolk sac. GCLm mRNA is constant throughout development, GCLc mRNA increases at gd 18 in both the placenta and the yolk sac. In the placenta the increase is localized to the spongiotrophoblast layer. GCLc protein level does not increase in parallel with the mRNA. The localization of GCLc mRNA and protein in mouse placenta is different, with the mRNA concentrated in the spongiotrophoblast and the protein in the labyrinth
-
catalytic subunit GCLc andmodifier subunit GCLm are differently regulated during development in the placenta and the yolk sac. GCLm mRNA is constant throughout development, GCLc mRNA increases at gd 18 in both the placenta and the yolk sac
additional information
-
male transgenic mice induced to overexpress glutamate-cysteine ligase exhibit resistance to acetaminophen-induced liver injury when compared with acetaminophen-treated male mice carrying, but not expressing glutamate-cysteine ligase transgenes, or to female glutamatecysteine ligase transgenic mice
additional information
-
correlating regional-specific expression and activity pattern, levels of heavy and light subunit are similar, distribution in brain
additional information
-
in most tissues the modifier subunit GCLM is limiting, suggesting that an increase in GCLM alone would increase gamma-glutamylcysteine synthesis
additional information
-
in most tissues the modifier subunit GCLM is limiting, suggesting that an increase in GCLM alone would increase gamma-glutamylcysteine synthesis
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
metabolism
P97494; O09172
certain carotenoids induce the Gcl mRNA expression in RAW264 cells and subsequently the GCL protein expression, which concomitantly enhances the intracellular GSH level, in a JNK pathway-related manner
physiological function
malfunction
-
erythrocytes from gclm-/- mice show greatly reduced intracellular glutathione. Prolonged incubation results in complete lysis of gclm-/- erythrocytes, which can be reversed by exogenous delivery of the antioxidant Trolox. Phenylhydrazine-induced oxidative stress in glcm-/- causes dramatically increased hemolysis, markedly larger accumulations of injured erythrocytes in the spleen, erythrocyte-derived pigment hemosiderin in kidney tubules, and diminished kidney function compared to wild-type mice, phenotype, overview. Regulatory subunit GCLM-deficient erythrocytes are more prone to Ca2+-dependent suicidal cell death ex vivo. Without additional oxidative stress, the mutant animals are able to survive by slightly ramping up their generation of new erythrocytes
malfunction
-
using GCLC knockout murine embryonic fibroblasts, addition of cysteine to catalytic subunit GCLC null cells results in a marked decrease in regulatory subunit GCLM mRNA levels despite the absence of GSH
physiological function
-
mice lacking the glutamate-cysteine ligase modifier subunit show an increase in myocardial ischaemia-reperfusion injury and apoptosis in ischaemic myocardium. A decrease in mitochondrial glutathione levels in ischaemic myocardium is more pronounced in mice lacking the glutamate-cysteine ligase modifier subunit than in control. The ESR signal intensity of the dimethyl-1-pyrroline-N-oxide-hydroxyl radical adducts in ischaemic myocardium is higher in mice lacking the glutamate-cysteine ligase modifier subunit than in control. Hypoxia-reoxygenation induces greater mitochondrial damage in cultured cardiomyocytes from mice lacking the glutamate-cysteine ligase modifier subunit
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
P97494; O09172
glutathione (GSH) is synthesized by a two-step enzyme reaction. In the first step, L-cysteine is ligated to the gamma carboxyl group of L-glutamic acid by glutamate-cysteine-ligase (GCL, EC 6.3.2.2), and in the second step L-glycine is bound to gamma-glutamylcysteine by glutathione synthase (EC 6.3.2.3). The first step is the ratelimiting step. Compared with the holoenzyme, the catalytic GCLc monomer shows lower enzymatic activity but higher sensitivity to feedback inhibition by GSH. Involvement of GCL activity in the increase in the intracellular GSH level induced by beta-carotene
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_MOUSE
637
0
72571
Swiss-Prot
other Location (Reliability: 4)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30500
1 * 72700, heavy catalytic subunit, + 1 * 30500, light regulatory subunit, SDS-PAGE
72700
1 * 72700, heavy catalytic subunit, + 1 * 30500, light regulatory subunit, SDS-PAGE
31000
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
1 * 72700, heavy catalytic subunit, + 1 * 30500, light regulatory subunit, SDS-PAGE
dimer
heterodimer
additional information
dimer
-
1 * 73000, about, catalytic subunit, + 1 * 31000, about, regulatory subunit, SDS-PAGE
dimer
-
heterodimer, comprising a catalytic subunit (GCLC) and a regulatory subunit (GCLM). GCLC alone can catalyze the formation of L-gamma-glutamyl-L-cysteine, its binding with GCLM enhances the enzyme activity by lowering the Km for glutamate and ATP, and increasing the Ki for GSH inhibition
heterodimer
-
glutamate-cysteine ligase consists of a catalytic subunit (GCLC) and a modifier subunit (GCLM)
heterodimer
P97494; O09172
glutamate-cysteine ligase (GCL) is a heterodimer enzyme composed of a catalytic subunit (GCLC) and a modulator subunit (GCLM)
additional information
-
the dimeric enzyme is composed of a heavy, catalytic subunit and a light, regulatory subunit
additional information
-
the holoenzyme consists of a heavy catalytic and a light regulatory subunit, i.e. gamma-GCSh and gamma-GCSl
additional information
-
reaction is catalyzed by the catalytic subunit GCLC or by the holoenzyme (GCLholo), which comprises GCLC and the modifier subunit GCLM. GCLM decreases the Km for ATP by about 6fold and decreases the Km-value for glutamate and increases the Ki-value for feedback inhibition by GSH. GCLM increases by 4.4fold the turnover number for gamma-glutamylcysteine synthesis
additional information
-
reaction is catalyzed by the catalytic subunit GCLC or by the holoenzyme (GCLholo), which comprises GCLC and the modifier subunit GCLM. GCLM decreases the Km for ATP by about 6fold and decreases the Km-value for glutamate and increases the Ki-value for feedback inhibition by GSH. GCLM increases by 4.4fold the turnover number for gamma-glutamylcysteine synthesis
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
-
the enzyme consists of a catalytic (GCLC) and a modifier (GCLM) subunit
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phosphoprotein
the enzyme exists in vivo as a mixture of phosphorylated and dephosphorylated forms
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
additional information
-
post-translational modifications of GCLC, e.g. phosphorylation, myristoylation, caspase-mediated cleavage, have modest effects on GCL activity
additional information
-
4-hydroxy-2-nonenal alters GCL holoenzyme formation and activity via direct posttranslational modification of the GCL subunits in vitro. 4-Hydroxy-2-nonenal directly modifies Cys553 of catalytic subunit GCLC and Cys35 of modulatory subunit GCLM in vitro, which significantly increases monomeric GCLC enzymatic activity, but reduces GCL holoenzyme activity and formation of the GCL holoenzymecomplex
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C248A/C249A
-
mutant enzyme shows the same strength of binding to regulatory subunit (GCLM) as does wild-type GCLC, yet the catalytic activity is dramatically decreased
E103A
-
transduction of Hepa-1c1c7 cells with a catalytically inactive GCL catalytic subunit E103A mutant decreases cellular GCL activity in a dose-dependent manner
P158L
-
mutant enzyme shows the same strength of binding to regulatory subunit (GCLM) as does wild-type GCLC, yet the catalytic activity is dramatically decreased
additional information
additional information
-
deletion analysis indicats that most regions, except a portion of the C-terminal region of catalytic subunit (GCLC) and a portion of the N-terminal region of regulatory subunit (GCLM), are required for the interaction to occur
additional information
-
construction of Gclc and Gclm transgenic mice designed to conditionally overexpress GCL in the liver, conditional Gcl transgene expression in these mice promotes resistance to acetaminophen-induced liver injury
additional information
-
transgenic mice that conditionally overexpress GCL subunits are protected from acetaminophen induced liver injury. Gclm null mice exhibit low GSH levels and enhanced sensitivity to acetaminophen. When Gclm expression and GCL activity are restored in Gclm conditional transgenic X Gclm null mice, they become resistant to APAP-induced liver damage. Construction of transgenic mouse models of inducible GCL subunit expression in the liver, overview
additional information
-
a protein transduction approach whereby recombinant GCL protein can be rapidly and directly transferred into cells when coupled to the HIV TAT protein transduction domain. The TAT-GCL fusion proteins are capable of heterodimerization and formation of functional GCL holoenzyme in vitro. Exposure of Hepa-1c1c7 cells to the TAT-GCL fusion proteins results in the time- and dose-dependent transduction of both GCL subunits and increased cellular GCL activity and glutathione levels. A heterodimerization-competent, enzymatically deficient GCLC-TAT mutant was also generated in an attempt to create a dominant-negative suppressor of GCL
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination and analysis, mapping to chromosome 9, band D-E, heavy and light subunits
generation of C57Bl/6 mice that conditionally overexpress glutamate-cysteine ligase
-
genes gclc and gclm, encoding the two different subunits, the different genes are located on separate chromosomes, quantitative RT-PCR enzyme expression analysis
P97494; O09172
Hepa-1 cells transfected with GCLC (catalytic subunit) and GCLM (modifier subunit) expression vectors
-
the 2 subunits are encoded by 2 different genes and are located on chromosomes 9D-E and 3HI-3, genetic regulation involving AP-1, constitutive expression, overview
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
actinomycin D and cycloheximide suppress enzyme expression. Using GCLC knockout murine embryonic fibroblasts, addition of cysteine to catalytic subunit GCLC null cells results in a marked decrease in regulatory subunit GCLM mRNA levels despite the absence of GSH. Addition of GSH similarly reduces GCLM mRNA abundance
-
As3+ coordinately upregulates GCL catalytic subunit and GCL modifier subunit mRNA levels resulting in increased GCL subunit protein expression, holoenzyme formation, and activity. As3+ increases the rate of transcription of both the GCL catalytic subunit and GCL modifier subunit genes and induces the posttranscriptional stabilization of GCL modifier subunit mRNA. The antioxidant N-acetylcysteine abolishes As3+-induced GCL catalytic subunit expression and attenuates induction of GCL modifier subunit. As3+ induction of GCL catalytic subunit and GCL modifier subunit is also differentially regulated by the MAPK signaling pathways and occurs independent of the Nrf1/2 transcription factors
-
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. The upregulation does not occur in MEFs lacking GCN2, i.e. general control non-derepressible 2, also known as eIF2a kinase 4, or in cells expressing mutant eIF2alpha lacking the eIF2alpha kinase Ser51 phosphorylation site, indicating that expression of both GCLC and GCLM is mediated by the GCN2/ATF4 stress response pathway
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enzyme induction in macrophages by beta-carotene or beta-cryptoxanthin. Both the protein and mRNA expression of GCL increases in a beta-carotene concentration-dependent manner. Buthionine sulfoximine, a GCL inhibitor, abolishes the beta-carotene-induced GSH increase without affecting the beta-carotene-induced GCL protein expression. Both cycloheximide, a translation inhibitor, and actinomycin D, a transcription inhibitor, completely suppressed the beta-carotene-induced GCL protein expression and the concomitant GSH increase. Similarly to beta-carotene, beta-cryptoxanthin upregulates the GCL protein expression, but lutein does not. The c-Jun N-terminal kinase (JNK) inhibitor, SP600125, suppresses the beta-carotene-induced GSH increase, whereas a p38 mitogen-activated protein kinase inhibitor or an extracellular signal-regulated kinase 1/2 inhibitor do not. The JNK inhibitor also suppresses the beta-carotene-induced GCL protein expression and consistently beta-carotene induced JNK phosphorylation
P97494; O09172
kaempferol protects cells against cisplatin-induced apoptosis in a dose-dependent manner in HEI-OC1 cells. Kaempferol increases the cellular level of glutathione and the expression of GCL catalytic subunit time-dependently. siRNA directed against GCL catalytic subunit blocks the increase of glutathione level by kaempferol and the protective effect of kaempferol against cisplatin-induced cell death
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
-
a protein transduction approach whereby recombinant GCL protein can be rapidly and directly transferred into cells when coupled to the HIV TAT protein transduction domain. The TAT-GCL fusion proteins are capable of heterodimerization and formation of functional GCL holoenzyme in vitro. Exposure of Hepa-1c1c7 cells to the TAT-GCL fusion proteins results in the time- and dose-dependent transduction of both GCL subunits and increased cellular GCL activity and glutathione levels. A heterodimerization-competent, enzymatically deficient GCLC-TAT mutant was also generated in an attempt to create a dominant-negative suppressor of GCL
medicine
medicine
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activation of glycogen synthase kinase 3beta is a key mediator of the initial phase of acetaminophen-induced liver injury through modulating GCL and Mcl-1 degradation, as well as JNK activation in liver. The silencing of glycogen synthase kinase 3beta decreases the loss of hepatic GCL, and promotes greater GSH recovery in liver following acetaminophen treatment
medicine
-
mice lacking the glutamate-cysteine ligase modifier subunit are susceptible to myocardial ischaemia-reperfusion injury partly through an increased vulnerability of mitochondria to oxidative damage owing to mitochondrial glutathione reduction
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Reid, L.L.; Botta, D.; Lu, Y.; Gallagher, E.P.; Kavanagh, T.J.
Molecular cloning and sequencing of the cDNA encoding the catalytic subunit of mouse glutamate-cysteine ligase
Biochim. Biophys. Acta
1352
233-237
1997
Mus musculus
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)
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
Kang, Y.; Viswanath, V.; Jha, N.; Qiao, X.; Mo, J.Q.; Andersen, J.K.
Brain gamma-glutamyl cysteine synthetase (GCS) mRNA expression patterns correlate with regional-specific enzyme activities and glutathione levels
J. Neurosci. Res.
58
436-441
1999
Mus musculus
Toroser, D.; Sohal, R.S.
Kinetic characteristics of native gamma-glutamylcysteine ligase in the aging housefly, Musca domestica L
Biochem. Biophys. Res. Commun.
326
586-593
2005
Mus musculus
Botta, D.; Franklin, C.C.; White, C.C.; Krejsa, C.M.; Dabrowski, M.J.; Pierce, R.H.; Fausto, N.; Kavanagh, T.J.
Glutamate-cysteine ligase attenuates TNF-induced mitochondrial injury and apoptosis
Free Radic. Biol. Med.
37
632-642
2004
Mus musculus
Chen, Y.; Shertzer, H.G.; Schneider, S.N.; Nebert, D.W.; Dalton, T.P.
Glutamate cysteine ligase catalysis: dependence on ATP and modifier subunit for regulation of tissue glutathione levels
J. Biol. Chem.
280
33766-33774
2005
Mus musculus
Diaz, D.; Krejsa, C.M.; White, C.C.; Charleston, J.S.; Kavanagh, T.J.
Effect of methylmercury on glutamate-cysteine ligase expression in the placenta and yolk sac during mouse development
Reprod. Toxicol.
19
117-129
2004
Mus musculus
Yang, Y.; Chen, Y.; Johansson, E.; Schneider, S.N.; Shertzer, H.G.; Nebert, D.W.; Dalton, T.P.
Interaction between the catalytic and modifier subunits of glutamate-cysteine ligase
Biochem. Pharmacol.
74
372-381
2007
Mus musculus
Toroser, D.; Yarian, C.S.; Orr, W.C.; Sohal, R.S.
Mechanisms of gamma-glutamylcysteine ligase regulation
Biochim. Biophys. Acta
1760
233-244
2006
Mus musculus (P97494), Mus musculus, Drosophila melanogaster (Q9W3K5), Drosophila melanogaster
Botta, D.; Shi, S.; White, C.C.; Dabrowski, M.J.; Keener, C.L.; Srinouanprachanh, S.L.; Farin, F.M.; Ware, C.B.; Ladiges, W.C.; Pierce, R.H.; Fausto, N.; Kavanagh, T.J.
Acetaminophen-induced liver injury is attenuated in male glutamate-cysteine ligase transgenic mice
J. Biol. Chem.
281
28865-28875
2006
Mus musculus
Nagashima, R.; Sugiyama, C.; Yoneyama, M.; Kuramoto, N.; Kawada, K.; Ogita, K.
Acoustic overstimulation facilitates the expression of glutamate-cysteine ligase catalytic subunit probably through enhanced DNA binding of activator protein-1 and/or NF-kappaB in the murine cochlea
Neurochem. Int.
51
209-215
2007
Mus musculus
Bea, F.; Hudson, F.N.; Neff-Laford, H.; White, C.C.; Kavanagh, T.J.; Kreuzer, J.; Preusch, M.R.; Blessing, E.; Katus, H.A.; Rosenfeld, M.E.
Homocysteine stimulates antioxidant response element-mediated expression of glutamate-cysteine ligase in mouse macrophages
Atherosclerosis
203
105-111
2008
Mus musculus
Botta, D.; White, C.C.; Vliet-Gregg, P.; Mohar, I.; Shi, S.; McGrath, M.B.; McConnachie, L.A.; Kavanagh, T.J.
Modulating GSH synthesis using glutamate cysteine ligase transgenic and gene-targeted mice
Drug Metab. Rev.
40
465-477
2008
Mus musculus
Wu, H.; White, C.C.; Isanhart, J.P.; McBride, T.J.; Kavanagh, T.J.; Hooper, M.J.
Optimization and application of glutamate cysteine ligase measurement in wildlife species
Ecotoxicol. Environ. Saf.
72
572-578
2008
Anas platyrhynchos, Mus musculus, Rattus norvegicus
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
Kobayashi, T.; Watanabe, Y.; Saito, Y.; Fujioka, D.; Nakamura, T.; Obata, J.E.; Kitta, Y.; Yano, T.; Kawabata, K.; Watanabe, K.; Mishina, H.; Ito, S.; Kugiyama, K.
Mice lacking the glutamate-cysteine ligase modifier subunit are susceptible to myocardial ischaemia-reperfusion injury
Cardiovasc. Res.
85
785-795
2010
Mus musculus
Thompson, J.A.; White, C.C.; Cox, D.P.; Chan, J.Y.; Kavanagh, T.J.; Fausto, N.; Franklin, C.C.
Distinct Nrf1/2-independent mechanisms mediate As 3+-induced glutamate-cysteine ligase subunit gene expression in murine hepatocytes
Free Radic. Biol. Med.
46
1614-1625
2009
Mus musculus
Shinohara, M.; Ybanez, M.D.; Win, S.; Than, T.A.; Jain, S.; Gaarde, W.A.; Han, D.; Kaplowitz, N.
Silencing glycogen synthase kinase-3beta inhibits acetaminophen hepatotoxicity and attenuates JNK activation and loss of glutamate cysteine ligase and myeloid cell leukemia sequence 1
J. Biol. Chem.
285
8244-8255
2010
Mus musculus
Gao, S.S.; Choi, B.M.; Chen, X.Y.; Zhu, R.Z.; Kim, Y.; So, H.; Park, R.; Sung, M.; Kim, B.R.
Kaempferol suppresses cisplatin-induced apoptosis via inductions of heme oxygenase-1 and glutamate-cysteine ligase catalytic subunit in HEI-OC1 cell
Pharm. Res.
27
235-245
2010
Mus musculus
Backos, D.S.; Brocker, C.N.; Franklin, C.C.
Manipulation of cellular GSH biosynthetic capacity via TAT-mediated protein transduction of wild-type or a dominant-negative mutant of glutamate cysteine ligase alters cell sensitivity to oxidant-induced cytotoxicity
Toxicol. Appl. Pharmacol.
243
35-45
2010
Mus musculus
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
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
Foeller, M.; Harris, I.S.; Elia, A.; John, R.; Lang, F.; Kavanagh, T.J.; Mak, T.W.
Functional significance of glutamate-cysteine ligase modifier for erythrocyte survival in vitro and in vivo
Cell Death Differ.
20
1350-1358
2013
Mus musculus
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