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Information on EC 6.3.2.2 - glutamate-cysteine ligase and Organism(s) Escherichia coli and UniProt Accession P0A6W9
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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: Escherichia coli
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
-
-
-
-
GCS
-
-
-
-
Synthetase, gamma-glutamylcysteine
-
-
-
-
gamma-Glutamylcysteine synthetase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine
reaction mechanism
-
ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine
reaction mechanism
ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine
no conserved cysteine residue in the active site
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 + alpha-ethyl-L-glutamate + L-alpha-aminobutyrate
ADP + phosphate + alpha-ethyl-L-glutamyl-L-alpha-aminobutyrate
-
-
ir
ATP + alpha-methyl-L-glutamate + L-alpha-aminobutyrate
ADP + phosphate + alpha-methyl-L-glutamyl-L-alpha-aminobutyrate
-
-
ir
ATP + D-Glu + L-alpha-aminobutyrate
ADP + phosphate + gamma-D-Glu-L-alpha-aminobutyrate
-
-
ir
ATP + L-Glu + allo-L-threonine
ADP + phosphate + gamma-L-Glu-allo-L-threonine
-
-
ir
ATP + L-Glu + beta-amino-iso-butyrate
ADP + phosphate + gamma-L-Glu-beta-amino-iso-butyrate
-
-
ir
ATP + L-Glu + beta-chloro-L-alanine
ADP + phosphate + gamma-L-Glu-beta-chloro-L-alanine
-
-
ir
ATP + L-Glu + beta-cyano-L-alanine
ADP + phosphate + gamma-L-Glu-beta-cyano-L-alanine
-
-
ir
ATP + L-Glu + L-2-aminobutanoate
ADP + phosphate + gamma-L-Glu-2-aminobutanoate
-
-
-
?
ATP + L-Glu + L-alanine
ADP + phosphate + gamma-L-Glu-L-alanine
-
-
ir
ATP + L-Glu + L-alpha-aminobutyrate
ADP + phosphate + gamma-L-Glu-L-alpha-aminobutyrate
-
-
ir
ATP + L-Glu + L-C-allylglycine
ADP + phosphate + gamma-L-Glu-L-C-allylglycine
-
-
ir
ATP + L-Glu + L-isoleucine
ADP + phosphate + gamma-L-Glu-L-isoleucine
-
-
ir
ATP + L-Glu + L-leucine
ADP + phosphate + gamma-L-Glu-L-leucine
-
-
ir
ATP + L-Glu + L-norleucine
ADP + phosphate + gamma-L-Glu-L-norleucine
-
-
ir
ATP + L-Glu + L-norvaline
ADP + phosphate + gamma-L-Glu-L-norvaline
-
-
ir
ATP + L-Glu + L-serine
ADP + phosphate + gamma-L-Glu-L-serine
-
-
ir
ATP + L-Glu + L-threonine
ADP + phosphate + gamma-L-Glu-L-threonine
-
-
ir
ATP + L-Glu + L-valine
ADP + phosphate + gamma-L-Glu-L-valine
-
-
ir
ATP + L-Glu + O-methyl-DL-serine
ADP + phosphate + gamma-L-Glu-O-methyl-DL-serine
-
-
ir
ATP + L-Glu + S-methyl-L-Cys
ADP + phosphate + gamma-L-Glu-L-S-methyl-Cys
-
-
ir
ATP + L-glutamate + L-cysteine
ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
-
-
?
ATP + N-methyl-L-glutamate + L-alpha-aminobutyrate
ADP + phosphate + N-methyl-L-glutamyl-L-alpha-aminobutyrate
-
-
ir
ATP + L-Glu + beta-chloro-L-Ala
ADP + phosphate + gamma-L-Glu-L-beta-chloro-L-Ala
-
strain KM: 79% of the activity relative to L-Cys, strain W: 99% of the activity relative to L-Cys
-
-
?
ATP + L-Glu + n-propylamine
ADP + phosphate + N-propyl-L-glutamine
-
-
-
-
?
ATP + L-Glu + S-methyl-L-Cys
ADP + phosphate + gamma-L-Glu-S-methyl-L-Cys
-
strain KM: 70% of the activity relative to L-Cys, strain W: 70% of the activity relative to L-Cys
-
-
?
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
-
-
ir
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
first and rate-limiting step in the GSH 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-2-aminobutanoate
ADP + phosphate + gamma-L-Glu-2-aminobutanoate
-
-
-
-
?
ATP + L-Glu + L-2-aminobutanoate
ADP + phosphate + gamma-L-Glu-2-aminobutanoate
-
strain KM: 85% of the activity relative to L-Cys, strain W: 81% of the activity relative to L-Cys
-
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
-
-
-
?
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
-
catalyzes the biosynthesis of the GSH precursor
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate-limiting step in the biosynthesis of glutathione
-
?
ATP + L-Glu + L-Ser
ADP + phosphate + gamma-L-Glu-L-Ser
-
strain KM: 18% of the activity relative to L-Cys, strain W: 13% of the activity relative to L-Cys
-
-
?
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 biosynthesis, plays a central role in glutathione homeostasis
-
-
?
additional information
?
-
substrate specificity, poor substrates are beta-glutamate, (R,S)-beta-methyl-DL-glutamate, (R,S)-gamma-methyl-glutamate, L-aspartate, and DL-alpha-aminoadipate
-
?
additional information
?
-
-
substrate specificity, poor substrates are beta-glutamate, (R,S)-beta-methyl-DL-glutamate, (R,S)-gamma-methyl-glutamate, L-aspartate, and DL-alpha-aminoadipate
-
?
additional information
?
-
-
enzyme is able to to combine glutamine and amines to form gamma-glutamylamides. The reaction rate depende on the length if the methylene chain of the amines in the following decreasing order: n-propylamine > butylamine > ethylamine > methylamine
-
-
?
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
-
-
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
first and rate-limiting step in the GSH 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 step in glutathione biosynthesis
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
catalyzes the biosynthesis of the GSH precursor
-
?
ATP + L-Glu + L-Cys
ADP + phosphate + gamma-L-Glu-L-Cys
-
first and rate-limiting step in the biosynthesis of glutathione
-
?
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 biosynthesis, plays a central role in glutathione homeostasis
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Cu2+
2 divalent metal ions per enzyme molecule are bound, can be replaced by Mn2+ and Mg2+, binding mechanism and kinetics, overview
Mg2+
Mn2+
2 divalent metal ions per enzyme molecule are bound, Mg2+ broadens the substrate specificity, decreases the resistance to L-buthionine-S,R-sulfoximine, can be replaced by Mg2+ and Cu2+, binding mechanism and kinetics, overview
Mg2+
Mg2+
2 divalent metal ions per enzyme molecule are bound, Mg2+ sharpens the substrate specificity, increases the resistance to L-buthionine-S,R-sulfoximine, can be replaced by Mn2+ and Cu2+, binding mechanism and kinetics, overview
Mg2+
-
divalent metal ion required, especially Mg2+, optimal concentration depending on ATP concentration
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(2S)-2-amino-4-[(2R,S)-2-carboxy-3-hydroxypropyl-(R,S)-sulfonimidoyl]butanoic acid
-
-
(2S)-2-amino-4-[(2R,S)-2-carboxy-3-phenylpropyl-(R,S)-sulfonimidoyl]butanoic acid
-
-
(2S)-2-amino-4-[(2R,S)-2-carboxypropyl-(R,S)-sulfonimidoyl]butanoic acid
-
-
(2S)-2-amino-4-[(2R,S)-2-carboxy-3-hydroxypropyl-(R,S)-sulfonimidoyl]butanoic acid
-
slow-binding, irreversible inactivation, ATP-dependent, a N-phosphorylated reaction intermediate is tightly bound to the enzyme, mechanism-based
(2S)-2-amino-4-[(2R,S)-2-carboxy-3-phenylpropyl-(R,S)-sulfonimidoyl]butanoic acid
-
weak, reversible inhibition
(2S)-2-amino-4-[(2R,S)-2-carboxybutyl-(R,S)-sulfonimidoyl]butanoic acid
-
slow-binding, irreversible inactivation, ATP-dependent, a N-phosphorylated reaction intermediate is tightly bound to the enzyme, mechanism-based
(2S)-2-amino-4-[(2R,S)-2-carboxyhexyl-(R,S)-sulfonimidoyl]butanoic acid
-
slow-binding, irreversible inactivation, ATP-dependent, a N-phosphorylated reaction intermediate is tightly bound to the enzyme, mechanism-based
(2S)-2-amino-4-[(2R,S)-2-carboxyoctyl-(R,S)-sulfonimidoyl]butanoic acid
-
weak, reversible inhibition
(2S)-2-amino-4-[(2R,S)-2-carboxypropyl-(R,S)-sulfonimidoyl]butanoic acid
-
slow-binding, irreversible inactivation, ATP-dependent, a N-phosphorylated reaction intermediate is tightly bound to the enzyme, mechanism-based
(2S)-2-amino-4-[2-carboxyethyl-(R,S)-sulfonimidoyl]butanoic acid
-
slow-binding, irreversible inactivation, ATP-dependent, a N-phosphorylated reaction intermediate is tightly bound to the enzyme, mechanism-based
L-buthionine-SR-sulfoximine
-
irreversible inactivation, ATP-dependent, a N-phosphorylated reaction intermediate is tightly bound to the enzyme, mechanism-based
L-glutamic acid gamma-monohydroxamate
-
ATP-dependent irreversible inactivation, loss of 90% activity within 3 days, inactivation mechanism, no inactivation occurs in absence of ATP or with AMP-PNP
L-buthionine-S-sulfoximine
mechanism-based inhibitor, in contrary to the mammalian enzyme form, the Escherichia coli enzyme is inhibited more weakly and slowly in presence of Mg2+, replacement of the metal by Mn2+ leads to increased binding affinity and inactivation rate
additional information
no inhibition by cysteamine or slowly at high concentration
-
additional information
-
no inacivationwith ATP alone or with L-aspartic acid gamma-monohydroxamate
-
additional information
-
the inhibition mode and potency of the different sulfoximines is highly dependent on the stereochemistry at the sulfoximine sulfur atom, overview
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00019
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
0.00024
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
0.00025
ATP
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
0.00053
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
0.0001
L-cysteine
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
0.00011
L-cysteine
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S
0.00011
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
0.00014
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
0.00016
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
0.0032
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
0.004
L-glutamate
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S
0.004
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
0.0041
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
0.0053
L-glutamate
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
additional information
additional information
Km values for diverse substrates in presence of Mg2+, or Mn2+, or both, kinetics
-
additional information
additional information
-
Km values for diverse substrates in presence of Mg2+, or Mn2+, or both, kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
24.2
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
27.2
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
30.1
ATP
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
30.5
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
21.9
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
22.2
L-cysteine
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
29.2
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
31.8
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
39.7
L-cysteine
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S
22.7
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
23.8
L-glutamate
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
30.4
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
31.2
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
34
L-glutamate
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
46.6
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
120.4
ATP
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
127.1
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
142.9
ATP
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
198.8
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
199.1
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
222
L-cysteine
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
288.6
L-cysteine
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
396.9
L-cysteine
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S
4.5
L-glutamate
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S, in presence of DTT
6.9
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
7.4
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
7.4
L-glutamate
-
pH 8.0, 25°C, mutant C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
8.5
L-glutamate
-
pH 8.0, 25°C, mutant C106S,C164S,C205S,C223S,C357S,C433S,C439S
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0089
(2S)-2-amino-4-[(2R,S)-2-carboxy-3-hydroxypropyl-(R,S)-sulfonimidoyl]butanoic acid
-
pH 7.5, 37°C
9.2
(2S)-2-amino-4-[(2R,S)-2-carboxy-3-phenylpropyl-(R,S)-sulfonimidoyl]butanoic acid
-
pH 7.5, 37°C
0.0000099 - 0.00059
(2S)-2-amino-4-[(2R,S)-2-carboxybutyl-(R,S)-sulfonimidoyl]butanoic acid
0.023
(2S)-2-amino-4-[(2R,S)-2-carboxyhexyl-(R,S)-sulfonimidoyl]butanoic acid
-
pH 7.5, 37°C
3.9
(2S)-2-amino-4-[(2R,S)-2-carboxyoctyl-(R,S)-sulfonimidoyl]butanoic acid
-
pH 7.5, 37°C
0.0014
(2S)-2-amino-4-[(2R,S)-2-carboxypropyl-(R,S)-sulfonimidoyl]butanoic acid
-
pH 7.5, 37°C
0.053
(2S)-2-amino-4-[2-carboxyethyl-(R,S)-sulfonimidoyl]butanoic acid
-
pH 7.5, 37°C
0.0000099
(2S)-2-amino-4-[(2R,S)-2-carboxybutyl-(R,S)-sulfonimidoyl]butanoic acid
-
pH 7.5, 37°C
0.00059
(2S)-2-amino-4-[(2R,S)-2-carboxybutyl-(R,S)-sulfonimidoyl]butanoic acid
-
pH 7.5, 37°C
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
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
gamma-glutamylcysteine synthetase (gamma-ECS) is a key enzyme in the biosynthesis pathway of glutathione (GSH), the precursor of phytochelatins. The overexpression of the bacterial gamma-glutamylcysteine synthetase in Populus tremula x Populus alba mediates changes in cadmium influx, allocation and detoxification in poplar, analysis of net Cd2+ influx in association with H+/Ca2+, Cd tolerance, and the underlying molecular and physiological mechanisms, overview. GSH-mediated induction of the transcription of genes involved in Cd2+ transport and detoxification
evolution
-
in eukaryotes gamma -glutamylcysteine synthetase and glutathione synthetase, EC 6.3.2.3, activities are encoded by two distinct enzymes In some prokaryotes, such as Escherichia coli and Vibrio cholerae, separate enzymes exist for these two reactions. However, in some prokaryotes, such as Streptococcus agalactiae, Pasteurella multicoda and Listeria monocytogenes, both of these activities are encoded by a single bifunctional enzyme, GshF. Evolution of gamma-GCS has occurred by convergent evolution in three different lineages with no significant sequence similarities between the lineages, the Escherichia coli enzyme belongs to lineage I
metabolism
-
biosynthesis of GSH occurs by two sequential ATP-dependent enzymatic steps. The first enzyme, gamma -glutamylcysteine synthetase ligates glutamate and cysteine to yield gamma -glutamylcysteine. Glutathione synthetase, EC 6.3.2.3, the second enzyme, then catalyses the addition of glycine to yield glutathione
physiological function
-
gamma-GCS is rate-limiting catalyzing the regulated step of GSH biosynthesis, being both transcriptionally and post-translationally regulated, post-translational regulation of the gamma-GCS enzyme by the redox environment
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
22.6 mg/ml purified enzyme, wild-type or mutant, hanging drop vapour diffusion method, equal volume of protein and reservoir solution, reservoir solution: 3.9 M sodium formate, pH 7.4, equilibration with reservoir solution at 293K, X-ray diffraction structure determination and analysis at 2.8 A resolution
-
sitting-drop vapor diffusion method, crystal structure of unliganded enzyme and enzyme complexed with a sulfoximine-based transition-state analog inhibitor at resolutions of 2.5 and 2.1 A, respectively. In the crystal structure of the complex, the bound inhibitor is phosphorylated at the sulfoximido nitrogen and is coordinated to three Mg2+ ions
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A494G
-
site-specific mutagenesis, 53% increased activity compared to wild-type enzyme
A494L
-
site-specific mutagenesis, 65% increased activity compared to wild-type enzyme
A494V
-
site-specific mutagenesis, 66% increased activity compared to wild-type enzyme
C 164S
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C106S
C106S/C164S/C205S/C223S
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S
-
site-directed mutagenesis, inactive mutant
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395W
-
site-directed mutagenesis, no complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/C372S/S395Y
-
site-directed mutagenesis, no complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q
-
site-directed mutagenesis, no complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/R374Q/V375F
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195, the mutant enzyme lacking cysteine residues shows a decreased in vivo half-life
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395W
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372C/S395Y
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372F/C395S
-
site-directed mutagenesis, no complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372F/S395C
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/S372W/S395C
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C395S/C433S/C439S/V375F
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C372S/C433S/C439S
-
site-directed mutagenesis, no complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C357S/C433S/C439S
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C106S/C164S/C205S/C223S/C433S/C439S
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C164S
-
site-directed mutagenesis, exchange of surface exposed cysteine residue for improved crystallization
C205S
C223S
C433S/C439S
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
S495T
-
site-specific mutagenesis, 62% increased activity compared to wild-type enzyme
additional information
C106S
-
site-directed mutagenesis, exchange of surface exposed cysteine residue for improved crystallization
C106S
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C205S
-
site-directed mutagenesis, exchange of surface exposed cysteine residue for improved crystallization
C205S
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
C223S
-
site-directed mutagenesis, exchange of surface exposed cysteine residue for improved crystallization
C223S
-
site-directed mutagenesis, complementation of the gcs yeast mutant strain ABC 1195
additional information
overexpression of Escherichia coli gamma-glutamylcysteine synthetase in the cytosol of Populus tremula x Populus alba produces higher glutathione (GSH) concentrations in leaves, thereby indicating the potential for cadmium (Cd) phytoremediation. Analysis of net Cd2+ influx in association with H+/Ca2+, Cd tolerance, and the underlying molecular and physiological mechanisms, overview. Transgenic plants have higher Cd2+ uptake rates and elevated transcript levels of several genes involved in Cd2+ transport and detoxification compared with wild-type poplar plants. Transgenic plants exhibit greater Cd2+ accumulation in the aerial parts than wild-type plants in response to Cd2+ exposure. Transgenic poplars show lower concentrations of superoxide anions and H2O2, higher concentrations of total thiols, GSH and oxidized GSH in roots and/or leaves, and stimulated foliar GSH reductase activity compared with wild-type plants. The transgenic plants are more tolerant of 0.1 mM Cd2+ than wild-type plants, probably due to the GSH-mediated induction of the transcription of genes involved in Cd2+ transport and detoxification
additional information
-
construction of a quadruple mutant of the enzyme termed gamma-GCS4CS
additional information
-
natural K-12 mutant B possesses a Gly at position 494 compared to Ala for the K-12 wild-type enzyme, the initiation codon exchange mutant shows increased activity
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
enzyme overexpression in Populus tremula x Populus alba (INRA female clone 717 1-B4) leaf cytosol, analysis of net Cd2+ influx in association with H+/Ca2+, Cd tolerance, and the underlying molecular and physiological mechanisms, overview
gene gshA, phylogenetic tree analysis of GCS lineage I, expression of C-terminally His-tagged wild-type and mutants in Escherichia coli strain BL21(DE3) or Origami from pTEF416, subcloning in Escherichia coli strain DH5alpha, complementation of Saccharomyces cerevisiae gsh deficient strain ABC1195
-
gene gshI with unusual initiation codon UUG, expression of wild-type and mutant enzymes in strain BH5262
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
agriculture
-
comparison of three transgenic poplar lines over-expressing the Escherichia coli gamma-glutamylcysteine synthetase. The three lines differ in their expression levels of the transgene and in the accumulation of gamma-glutamylcysteine and glutathione in leaves, roots and phloem exudates. The lowest transgene expression level is observed in line Lggs6 which shows an increased growth, an enhanced rate of photosynthesis and a decreased excitation pressure. Line Lggs12 shows the highest transgene expression level, highest gamma-glutamylcysteine accumulation in leaves and highest glutathione enrichment in phloem exudates and roots. This line also exhibits a reduced growth, and after a prolonged growth of 4.5 months, symptoms of leaf injury
medicine
-
the enzyme is a target for development of potential therapeutic agents
synthesis
-
resting cells of Escherichia coli expressing gamma-glutamylcysteine synthetase, with ATP regeneration through glycolysis, synthesize 12.1 mM theanine in 18 h from 429 mM ethylamine
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Huang, C.S.; Moore, W.R.; Meister, A.
On the active site thiol of gamma-glutamylcysteine synthetase: relationship to catalysis, inhibition, and regulation
Proc. Natl. Acad. Sci. USA
85
2464-2468
1988
Escherichia coli, Rattus norvegicus
Watanabe, K.; Murata, K.; Kimura, A.
Purification and characterization of gamma-glutamylcysteine synthetase of Escherichia coli B
Agric. Biol. Chem.
50
1925-1930
1986
Escherichia coli, Escherichia coli B / ATCC 11303
-
Kumagai, H.; Nakayama, R.; Tochikura, T.
gamma-Glutamylcysteine synthetase from Proteus mirabilis
Agric. Biol. Chem.
46
1301-1309
1982
Bacterium cadaveris, Escherichia coli, Escherichia coli B / ATCC 11303, Escherichia coli Crooks, Escherichia coli FKU-1, Escherichia coli FKU-3, Escherichia coli FKU-8, Escherichia coli K-10, Escherichia coli S-96, Klebsiella aerogenes, Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas schuylkilliensis
-
Murata, K.; Kimura, A.
Cloning of a gene responsible for the biosynthesis of glutathione in Escherichia coli B
Appl. Environ. Microbiol.
44
1444-1448
1982
Escherichia coli, Escherichia coli B / ATCC 11303
Inoue, Y.; Iba, Y.; Yano, H.; Murata, K.; Kimura, A.
Functional analysis of the gamma-glutamylcysteine synthetase of Escherichia coli B: effect of substitution of His-150 to Ala
Appl. Microbiol. Biotechnol.
38
473-477
1993
Escherichia coli, Escherichia coli B / ATCC 11303
Hibi, T.; Hisada, H.; Nakatsu, T.; Kato, H.; Oda, J.
Escherichia coli B gamma-glutamylcysteine synthetase: modification, purification, crystallization and preliminary crystallographic analysis
Acta Crystallogr. Sect. D
58
316-318
2002
Escherichia coli, Escherichia coli B / ATCC 11303
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)
Katoh, M.; Hiratake, J.; Oda, J.i.
ATP-dependent inactivation of Escherichia coli gamma-glutamylcysteine synthetase by L-glutamic acid gamma-monohydroxamate
Biosci. Biotechnol. Biochem.
62
1455-1457
1998
Escherichia coli
Hiratake, J.; Irie, T.; Tokutake, N.; Oda, J.i.
Recognition of a cysteine substrate by E. coli gamma-glutamylcysteine synthetase probed by sulfoximine-based transition-state analogue inhibitors
Biosci. Biotechnol. Biochem.
66
1500-1514
2002
Escherichia coli
Kwak, J.H.; Nam, Y.S.; Lee, S.Y.
Site-specific mutagenesis of the gshI gene for increasing the activity of gamma-glutamylcysteine synthetase in Escherichia coli K-12
J. Biochem. Mol. Biol.
31
254-257
1998
Escherichia coli
-
Kelly, B.S.; Antholine, W.E.; Griffith, O.W.
Escherichia coli gamma-glutamylcysteine synthetase. Two active site metal ions affect substrate and inhibitor binding
J. Biol. Chem.
277
50-58
2002
Escherichia coli (P0A6W9), Escherichia coli, Escherichia coli JM109 (P0A6W9)
Hibi, T.; Nii, H.; Nakatsu, T.; Kimura, A.; Kato, H.; Hiratake, J.; Oda, J.
Crystal structure of gamma-glutamylcysteine synthetase: insights into the mechanism of catalysis by a key enzyme for glutathione homeostasis
Proc. Natl. Acad. Sci. USA
101
15052-15057
2004
Escherichia coli
Miyake, K.; Kakita, S.
A novel catalytic ability of gamma-glutamylcysteine synthetase of Escherichia coli and its application in theanine production
Biosci. Biotechnol. Biochem.
73
2677-2683
2009
Escherichia coli
Herschbach, C.; Rizzini, L.; Mult, S.; Hartmann, T.; Busch, F.; Peuke, A.D.; Kopriva, S.; Ensminger, I.
Over-expression of bacterial gamma-glutamylcysteine synthetase (GSH1) in plastids affects photosynthesis, growth and sulphur metabolism in poplar (Populus tremula x Populus alba) dependent on the resulting gamma-glutamylcysteine and glutathione levels
Plant Cell Environ.
33
1138-1151
2010
Escherichia coli
Kumar, S.; Kasturia, N.; Sharma, A.; Datt, M.; Bachhawat, A.K.
Redox-dependent stability of the gamma-glutamylcysteine synthetase enzyme of Escherichia coli: a novel means of redox regulation
Biochem. J.
449
783-794
2013
Escherichia coli
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