Information on EC 6.3.2.3 - Glutathione synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
6.3.2.3
-
RECOMMENDED NAME
GeneOntology No.
Glutathione synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione
show the reaction diagram
the catalytic mechanism is proposed to proceed via phosphorylation of the dipeptide substrate to yield an acyl phosphate intermediate. This intermediate is subsequently attacked by glycine, followed by loss of inorganic phosphate, leading to glutathione formation
-
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione
show the reaction diagram
determination of reaction and substrate binding mechanisms, large conformational changes in the catalytic cycle
Q08220
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione
show the reaction diagram
mechanism, active site residues are Glu144, Asn146, Lys305, and Lys364, interaction of these residues with the ligands is essential for enzyme activity, especially Glu144 seems to be very important for stabilization of the reaction intermediate, but the active site residues are not essential for the overall enzyme structure
-
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione
show the reaction diagram
residues A534 and A535 are involved in substrate binding and determination of substrate specificity
Q94GA0
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione
show the reaction diagram
substrate binding kinetics and mechanism, negative cooperativity of gamma-L-Glu-L-Cys
-
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione
show the reaction diagram
random ter-reactant reaction mechanism, overview
P73493
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione
show the reaction diagram
the reaction process involves the formation of an acyl phosphate on the cysteinyl moiety in L-gamma-glutamyl-L-cysteine, followed by the attack of the glycine and formation of an enzyme-product complex, which finally dissociates with the release of GSH, ADP and phosphate
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
carboxamide formation
-
-
-
-
carboxylic acid-amide formation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
glutathione biosynthesis
-
-
Glutathione metabolism
-
-
glutathione metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
gamma-L-Glutamyl-L-cysteine:glycine ligase (ADP-forming)
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
gamma -glutamate-cysteine ligase-glutathione synthetase
-
bifunctional enzyme
gamma-GCS-GS
-
bifunctional enzyme accounts for gamma-glutamylcysteine synthetase and glutathione synthetase activities
gamma-GCS-GS
-
bifunctional enzyme, with a glutathione synthetase at the C terminus
gamma-glutamate-cysteine ligase/glutathione synthetase
-
-
gamma-glutamylcysteine synthetase-glutathione synthetase
-
-
GCL
P73493
-
Glutathione synthase
-
-
-
-
Glutathione synthase
-
-
Glutathione synthetase
-
-
-
-
Glutathione synthetase
-
-
Glutathione synthetase
P46416
-
Glutathione synthetase
-
-
Glutathione synthetase
-
-
Glutathione synthetase
Clostridium acetobutylicum NBRC13948
-
-
-
Glutathione synthetase
-
-
Glutathione synthetase
-
-
Glutathione synthetase
-
-
Glutathione synthetase
Q94GA0
-
Glutathione synthetase
-
-
Glutathione synthetase
-
-
Glutathione synthetase
Q9FV26
-
Glutathione synthetase
Q9FVA2
-
Glutathione synthetase
-
-
Glutathione synthetase
AF333982
-
Glutathione synthetase
P46413
-
Glutathione synthetase
-
-
Glutathione synthetase
Rhizobium tropici CIAT899
-
-
-
Glutathione synthetase
-
-
Glutathione synthetase
P35669
-
Glutathione synthetase
Q8DXM9
-
Glutathione synthetase
Streptococcus agalactiae 2603
Q8DXM9
-
-
Glutathione synthetase
D7P1H2
bifunctional enzyme, also gamma-glutamylcysteine ligase activity, EC 6.3.2.2
Glutathione synthetase
-
-
Glutathione synthetase
-
-
Glutathione synthetase (tripeptide)
-
-
-
-
GS
AF333982
-
GSH synthetase
-
-
-
-
GSH-S
-
-
GSH2
-
-
GSHase
-
-
-
-
gshB
P73493
gene name
GshF
-
bifunctional enzyme; bifunctional enzyme gamma -glutamate-cysteine ligase-glutathione synthetase
GSHS
Q9FV26
-
GSHS
Q9FVA2
-
GSHS1
Q94GA0
-
GSS
-
-
-
-
Phytochelatin synthetase
-
-
-
-
Synthetase, glutathione
-
-
-
-
TAGS1
Q7X9G1
-
TaGS2
Q7X9G0
-
ZmGS
Q7XB40
-
GSS
P46413
-
additional information
-
the enzyme belongs to the glutathione synthetase ATP-binding domain-like superfamily
additional information
-
enzyme belongs to the ATP-grasp enzyme superfamily
CAS REGISTRY NUMBER
COMMENTARY
9023-62-5
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
A. thaliana
-
-
Manually annotated by BRENDA team
A. thaliana; wild-type and deletion mutants: DELTA67-71 and DELTA67-200
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes
-
-
Manually annotated by BRENDA team
gene gsh2
UniProt
Manually annotated by BRENDA team
indian mustard
-
-
Manually annotated by BRENDA team
Brettanomyces abstinens
-
-
-
Manually annotated by BRENDA team
strain NBRC13948
-
-
Manually annotated by BRENDA team
Clostridium acetobutylicum NBRC13948
strain NBRC13948
-
-
Manually annotated by BRENDA team
B; Escherichia coli B enzyme overproduced in Escherichia coli JM109
-
-
Manually annotated by BRENDA team
B; wild-type and loopless mutant, in which the loop (Ile226-Arg241) is replaced with 3 Gly residues
-
-
Manually annotated by BRENDA team
B; wild-type and mutant enzymes
-
-
Manually annotated by BRENDA team
gene gshII
-
-
Manually annotated by BRENDA team
JM109, wild-type, deletion mutant of the multifunctional loop and mutant with a nicked loop
-
-
Manually annotated by BRENDA team
K12, strain AB 1157
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes with Cys residues replaced by Ala
-
-
Manually annotated by BRENDA team
recombinant
SwissProt
Manually annotated by BRENDA team
wild-type and mutant enzymes: Cys294Ala, Cys409Ala, Cys422Ala and Cys294/Cys409/Cys422 to Ala294/Ala409/Ala422
-
-
Manually annotated by BRENDA team
gene gshs1, cv. Jemalong genotype J5
SwissProt
Manually annotated by BRENDA team
normal and ApoE-/- mice
-
-
Manually annotated by BRENDA team
swiss mice
-
-
Manually annotated by BRENDA team
isozyme GSHS2, strain 3622
SwissProt
Manually annotated by BRENDA team
Pigeon
-
-
-
Manually annotated by BRENDA team
isozyme GSHS1
SwissProt
Manually annotated by BRENDA team
isozyme GSHS2
SwissProt
Manually annotated by BRENDA team
cold-adapted, gene gshB
UniProt
Manually annotated by BRENDA team
cold-adapted, gene gshB
UniProt
Manually annotated by BRENDA team
-
AF333982
GenBank
Manually annotated by BRENDA team
male sprague-dawley rats
-
-
Manually annotated by BRENDA team
Sprague-Dawley rats
-
-
Manually annotated by BRENDA team
Sprague-Dawley rats
UniProt
Manually annotated by BRENDA team
strain CIAT899
-
-
Manually annotated by BRENDA team
Rhizobium tropici CIAT899
strain CIAT899
-
-
Manually annotated by BRENDA team
Saccharomyces pombe
-
-
-
Manually annotated by BRENDA team
mutant MN101 transformed with pYS41 containing the glutathione synthetase gene
-
-
Manually annotated by BRENDA team
strain 2603 V/R
UniProt
Manually annotated by BRENDA team
Streptococcus agalactiae 2603
strain 2603 V/R
UniProt
Manually annotated by BRENDA team
bifucntional glutamate-cysteine ligase and glutathione synthetase
UniProt
Manually annotated by BRENDA team
Streptococcus thermophilus SIIM B218
bifucntional glutamate-cysteine ligase and glutathione synthetase
UniProt
Manually annotated by BRENDA team
gene gshB or slr1238
UniProt
Manually annotated by BRENDA team
recombinant
SwissProt
Manually annotated by BRENDA team
cowpea, strain 32H1
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
recombinant
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
P35669
homoglutathione synthetase, EC 6.3.2.23, has evolved from glutathione synthetase by a single gene duplication event
evolution
P73493
Synechocystis glutathione synthase shares properties with other prokaryotic enzymes
physiological function
P73493
isolation of fully segregated gshB deletion mutants. The mutant strain lacks reduced glutathione but instead accumulates the precursor compound gamma-glutamylcysteine. The deletion strain grows slower than the wild-type strain under favorable conditions and exhibits extremely reduced growth or death when subjected to conditions promoting oxidative stress. After subjecting the strains to multiple environmental and redox perturbations, we found that conditions promoting growth stimulate glutathione biosynthesis. Cellular GSH and gamma-glutamylcysteine content decline following exposure to dark and blue light and during photoheterotrophic growth. A rapid depletion of GSH and gamma-glutamylcysteine is observed in the wild type and the mutant strain, when cells are starved for nitrate or sulfate
physiological function
P73493
glutathione biosynthesis catalysed by glutamate-cysteine ligase, EC 6.3.2.2, and glutathione synthetase is essential for maintaining redox homoeostasis and protection against oxidative damage in diverse eukaroytes and bacteria
metabolism
P35669
A485L/T486P mutant shows a shift the substrate specificity increased affinity of GSH2 for Ser as a substrate, while affinity to Gly is preserved. This provides a new biosynthetic pathway for hydroxymethyl glutathione, which is known to be synthesized from glutathione and Ser in a reaction catalysed by carboxypeptidase Y
additional information
P35669
structure comparisons and homology structure molecular modelling of the GSH2 wild-type and mutant enzymes, overview
additional information
-
the active site is composed of three highly conserved catalytic loops, notably the alanine rich A-loop, Asp458 is important for cooperativity and active site structure, it impacts the allostery of the enzyme. Asp458 is important for loop closure and is a critical residue within the enzyme's A-loop. Enzyme structure-function modeling, molecular dynamics, overview
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + acetyl-Cys + Gly
ADP + phosphate + acetyl-Cys-Gly
show the reaction diagram
-
-
-
-
-
ATP + beta-aminoglutaryl-L-2-aminobutyrate + Gly
ADP + phosphate + beta-aminoglutaryl-L-2-aminobutyryl-Gly
show the reaction diagram
-
-
-
-
-
ATP + beta-aminoglutaryl-L-2-aminobutyrate + Gly
ADP + phosphate + beta-aminoglutaryl-L-2-aminobutyryl-Gly
show the reaction diagram
-
15% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + DL-gamma-(beta-methyl)Glu-L-2-aminobutyrate + Gly
ADP + phosphate + DL-gamma-(beta-methyl)Glu-L-2-aminobutyryl-Gly
show the reaction diagram
-
44% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + DL-gamma-(gamma-methyl)Glu-L-2-aminobutyrate + Gly
ADP + phosphate + DL-gamma-(gamma-methyl)Glu-L-2-aminobutyryl-Gly
show the reaction diagram
-
16% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + gamma-(alpha-aminomethyl)Glu-2-aminobutyrate + Gly
ADP + phosphate + gamma-Glu-L-2-aminopentanoyl-Gly
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-2-aminobutyrate + hydroxylamine
ADP + phosphate + gamma-(alpha-aminomethyl)Glu-2-aminobutyryl-Gly
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-aminobutyrate + Gly
ADP + phosphate + ophthalmic acid
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-aminobutyrate + Gly
ADP + phosphate + ophthalmic acid
show the reaction diagram
-
-
-
-
ATP + gamma-Glu-aminobutyrate + Gly
ADP + phosphate + ophthalmic acid
show the reaction diagram
-
-
-
-
ATP + gamma-Glu-aminobutyrate + Gly
ADP + phosphate + ophthalmic acid
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-aminobutyrate + Gly
ADP + phosphate + ophthalmic acid
show the reaction diagram
-
L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + gamma-Glu-aminobutyrate + Gly
ADP + phosphate + ophthalmic acid
show the reaction diagram
-
D-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + gamma-Glu-L-Cys + Ala
ADP + phosphate + gamma-Glu-Cys-Ala
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Ala
ADP + phosphate + gamma-Glu-Cys-Ala
show the reaction diagram
-
beta-alanine
-
-
-
ATP + gamma-Glu-L-Cys + aminomethanesulfonic acid
ADP + phosphate + gamma-Glu-L-Cys-aminomethanesulfonic acid
show the reaction diagram
Pigeon
-
-
-
-
-
ATP + gamma-Glu-L-Cys + aminooxyacetate
ADP + phosphate + gamma-Glu-Cys-aminoxyacetate
show the reaction diagram
-
53% of the activity relative to Gly
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
P04425
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q8W4W2
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Pigeon
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Saccharomyces pombe
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q8DXM9
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
i.e. gamma-Glu-Cys-Gly
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
i.e. gamma-Glu-Cys-Gly
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
i.e. gamma-Glu-Cys-Gly
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
i.e. gamma-Glu-Cys-Gly
-
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
patients with hereditary glutathione synthetase deficiency suffer from haemolytic anaemia, 5-oxoprolinuria, metabolic acidosis, recurrent bacterial infections and various degrees of central nervous system dysfunction
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
two NFE2 sites in the human GSS promoter play important roles in the basal expression of glutathione synthetase. The glutathione synthetase gene expression is also regulated by Nrf2
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
P46416
random ter-reactant mechanism, ADP is the last product released
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Streptococcus agalactiae 2603
Q8DXM9
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Clostridium acetobutylicum NBRC13948
-
-
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + gamma-glutathione
show the reaction diagram
-
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + gamma-glutathione
show the reaction diagram
Q9FV27, Q9FVA2
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + gamma-glutathione
show the reaction diagram
Q9FV26
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
-
glutathione biosynthesis
-
-
-
ATP + gamma-Glu-L-Cys + N-hydroxyglycine
ADP + phosphate + gamma-Glu-Cys-N-hydroxyglycine
show the reaction diagram
-
33% of the activity relative to Gly
-
-
-
ATP + gamma-Glu-L-Cys + Ser
ADP + phosphate + gamma-Glu-Cys-Ser
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-S-methylcysteine + Gly
ADP + phosphate + gamma-Glu-S-methylcysteinyl-Gly
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-S-methylcysteine + Gly
ADP + phosphate + gamma-Glu-S-methylcysteinyl-Gly
show the reaction diagram
-
24% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + gamma-glutamyl-alpha-aminobutyrate
?
show the reaction diagram
-
-
-
?
ATP + gamma-L-Glu-aminobutanoate + Gly
ADP + phosphate + ophthalmic acid
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-Glu-L-alpha-aminobutyrate + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
r
ATP + gamma-L-Glu-L-Cys + 3-amino-1-propanol
ADP + phosphate + L-gamma-glutamyl-N-(3-hydroxypropyl)-L-cysteinamide
show the reaction diagram
Q9M426
-
-
-
?
ATP + gamma-L-Glu-L-Cys + 3-amino-1-propanol
ADP + phosphate + L-gamma-glutamyl-N-(3-hydroxypropyl)-L-cysteinamide
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + alpha-aminobutyric acid
ADP + phosphate + gamma-Glu-L-Cys-alpha-aminobutyric acid
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + beta-Ala
ADP + phosphate + gamma-Glu-L-Cys-beta-Ala
show the reaction diagram
Q7XB40
-
-
-
?
ATP + gamma-L-Glu-L-Cys + beta-Ala
ADP + phosphate + gamma-Glu-L-Cys-beta-Ala
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + beta-Ala
ADP + phosphate + gamma-Glu-L-Cys-beta-Ala
show the reaction diagram
Q9M426
no activity with L-Ala or D-Ala
-
-
?
ATP + gamma-L-Glu-L-Cys + beta-Ala
ADP + phosphate + gamma-L-Glu-L-Cys-beta-Ala
show the reaction diagram
Q7X9G0, Q7X9G1
no activity with D-Ala
-
-
?
ATP + gamma-L-Glu-L-Cys + beta-aminoisobutyric acid
ADP + phosphate + gamma-Glu-L-Cys-beta-aminoisobutyric acid
show the reaction diagram
Q9M426
-
-
-
?
ATP + gamma-L-Glu-L-Cys + beta-aminoisobutyric acid
ADP + phosphate + gamma-Glu-L-Cys-beta-aminoisobutyric acid
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + beta-aminoisobutyric acid
ADP + phosphate + gamma-L-Glu-L-Cys-beta-aminoisobutyric acid
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + D-Ala
ADP + phosphate + gamma-Glu-L-Cys-D-Ala
show the reaction diagram
Q7XB40
-
-
-
?
ATP + gamma-L-Glu-L-Cys + D-Ala
ADP + phosphate + gamma-Glu-L-Cys-D-Ala
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + D-Ser
ADP + phosphate + gamma-Glu-L-Cys-D-Ser
show the reaction diagram
Q9M426
-
-
-
?
ATP + gamma-L-Glu-L-Cys + D-Ser
ADP + phosphate + gamma-Glu-L-Cys-D-Ser
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + ethanolamine
ADP + phosphate + gamma-Glu-L-Cys-ethanolamine
show the reaction diagram
Q9M426
-
-
-
?
ATP + gamma-L-Glu-L-Cys + ethanolamine
ADP + phosphate + gamma-Glu-L-Cys-ethanolamine
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + gamma-aminobutyric acid
ADP + phosphate + gamma-Glu-L-Cys-gamma-aminobutyric acid
show the reaction diagram
Q7XB40
-
-
-
?
ATP + gamma-L-Glu-L-Cys + gamma-aminobutyric acid
ADP + phosphate + gamma-Glu-L-Cys-gamma-aminobutyric acid
show the reaction diagram
Q9M426
-
-
-
?
ATP + gamma-L-Glu-L-Cys + gamma-aminobutyric acid
ADP + phosphate + gamma-Glu-L-Cys-gamma-aminobutyric acid
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + gamma-aminobutyric acid
ADP + phosphate + gamma-L-Glu-L-Cys-gamma-aminobutyric acid
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q08220
-
-
ir
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q94GA0
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
AF333982
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q7XB40
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q9M426
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
cells are not able to grow without glutathione, wild-type and all mutant enzyme forms can restore activity of the deficient mutant strain on minimal medium without glutathione
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
gamma-glutamyl-cysteine is bound cooperatively by the enzyme, ATP might play a key role in sequential binding, but its amount does not alter the cooperative binding of gamma-glutamyl-cysteine
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
in presence of dithiothreitol
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
inpresence of dithiothreitol
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
P48637
wild-type enzyme shows negative cooperativity
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
wild-type enzyme shows negative cooperativity in binding of gamma-L-Glu-L-Cys
-
r
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
ApoE-deficient mice show reduced enzyme expression and activity, genetic regulation mechanism, overview
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q08220
final step in glutathione biosynthesis
-
ir
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
final step of glutathione biosynthesis
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
gene is regulated by stresses via the Atf1-Spc-1-Wis1 signal pathway
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
P48637
second step in the biosynthesis of glutahione
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
second step in the biosynthesis of glutahione
-
r
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
enzyme catalyzes the final step of glutathione biosynthesis
-
-
?
ATP + gamma-L-Glu-L-Cys + L-2,3-diaminopropionic acid
ADP + phosphate + gamma-Glu-L-Cys-L-2,3-diaminopropionic acid
show the reaction diagram
Q7XB40
-
-
-
?
ATP + gamma-L-Glu-L-Cys + L-2,3-diaminopropionic acid
ADP + phosphate + gamma-Glu-L-Cys-L-2,3-diaminopropionic acid
show the reaction diagram
Q9M426
-
-
-
?
ATP + gamma-L-Glu-L-Cys + L-2,3-diaminopropionic acid
ADP + phosphate + gamma-Glu-L-Cys-L-2,3-diaminopropionic acid
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + L-2,3-diaminopropionic acid
ADP + phosphate + gamma-L-Glu-L-Cys-L-2,3-diaminopropionic acid
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + L-Ala
ADP + phosphate + gamma-Glu-L-Cys-L-Ala
show the reaction diagram
Q7XB40
-
-
-
?
ATP + gamma-L-Glu-L-Cys + L-Ala
ADP + phosphate + gamma-Glu-L-Cys-L-Ala
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-Glu-L-Cys + L-Ala
ADP + phosphate + gamma-L-Glu-L-Cys-L-Ala
show the reaction diagram
Q7X9G0, Q7X9G1
no activity with D-Ala
-
-
?
ATP + gamma-L-Glu-L-Cys + L-Orn
ADP + phosphate + gamma-Glu-L-Cys-L-Orn
show the reaction diagram
Q9M426
-
-
-
?
ATP + gamma-L-Glu-L-Cys + L-Orn
ADP + phosphate + gamma-Glu-L-Cys-L-Orn
show the reaction diagram
Q7X9G0, Q7X9G1
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P46413
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P46416
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P73493
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
Q3IFA2
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P35669
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
an optimally functioning two-step glutathione biosynthetic pathway is required in vivo for a robust defense against arsenite
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
glutathione is a ubiquitous intracellular peptide with diverse functions that include detoxification, antioxidant defense, maintenance of thiol status, and modulation of cell proliferation, whose biosynthesis is tightly regulated, overview. GSH synthase is regulated in a coordinated manner and its up-regulation can further enhance the capacity of the cell to synthesize GSH, enzyme regulation, detailed overview
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
glutathione is a ubiquitous intracellular peptide with diverse functions that include detoxification, antioxidant defense, maintenance of thiol status, and modulation of cell proliferation, whose biosynthesis is tightly regulated, overview. GSH synthase is regulated in a coordinated manner and its up-regulation can further enhance the capacity of the cell to synthesize GSH, enzyme regulation, detailed overview. Changes in GSH homeostasis occur in mice with liver-specific retinoid X receptor RXRalpha deletion
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P46416
GSH homeostasis in plants is essential for cellular redox control and efficient responses to abiotic and biotic stress. Compartmentation of the GSH biosynthetic pathway is a unique feature of plants, restricting glutathione biosynthesis to the cytosol is sufficient for normal plant development, overview
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P46413
the enzyme catalyzes the last step in glutathione biosynthesis
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
the enzyme catalyzes the last step in glutathione biosynthesis, loss of intracellular neuronal GSH is an important feature of neurodegenerative disorders including Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis, overview
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
Q3IFA2
-
-
-
?
ATP + Glu-L-2-aminobutyrate + Gly
ADP + phosphate + gamma-Glu-L-Cys-Gly
show the reaction diagram
-
16% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + L-gamma-(alpha-methyl)Glu-L-2-aminobutyrate + Gly
ADP + phosphate + L-gamma-(alpha-methyl)Glu-L-2-aminobutyryl-Gly
show the reaction diagram
-
-
-
-
-
ATP + L-gamma-(N-methyl)Glu-L-2-aminobutyrate + Gly
ADP + phosphate + L-gamma-(N-methyl)Glu-L-2-aminobutyryl-Gly
show the reaction diagram
-
52% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + L-gamma-Glu-Gly + Gly
ADP + phosphate + L-gamma-Glu-Gly-Gly
show the reaction diagram
-
8% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + L-gamma-Glu-L-2-aminopentanoic acid + Gly
ADP + phosphate + gamma-Glu-S-methylcysteinyl-Gly
show the reaction diagram
-
9% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + L-gamma-Glu-L-Ala + Gly
ADP + phosphate + L-gamma-Glu-L-Ala-Gly
show the reaction diagram
-
55% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + L-gamma-Glu-L-Cys + Gly
ADP + phosphate + L-gamma-Glu-L-Cys-Gly
show the reaction diagram
-
102% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + L-gamma-Glu-L-Ser + Gly
ADP + phosphate + L-gamma-Glu-L-Ser-Gly
show the reaction diagram
-
77% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
ATP + N-acetyl-L-2-aminobutyrate + Gly
ADP + phosphate + N-acetyl-L-2-aminobutyrate-Gly
show the reaction diagram
-
8% of the activity relative to L-gamma-glutamyl-L-2-aminobutyrate
-
-
-
ATP + N-acetyl-L-Cys + Gly
ADP + phosphate + N-acetyl-L-Cys-Gly
show the reaction diagram
-
20% of the activity relative to L-gamma-Glu-L-2-aminobutyrate
-
-
-
CTP + gamma-Glu-L-Cys + Gly
CDP + phosphate + glutathione
show the reaction diagram
-
4% of the activity relative to ATP
-
-
-
dATP + gamma-Glu-L-Cys + Gly
dADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
-
dATP + gamma-Glu-L-Cys + Gly
dADP + phosphate + glutathione
show the reaction diagram
-
75% of the activity relative to ATP
-
-
-
gamma-L-glutamyl-L-cysteine + glycine + ATP
ADP + phosphate + glutathione
show the reaction diagram
D7P1H2
-
-
-
?
glycine + ATP + gamma-L-glutamyl-L-cysteine
ADP + phosphate + glutathione
show the reaction diagram
-
assay at pH 8.2
-
-
?
GTP + gamma-Glu-L-Cys + Gly
GDP + phosphate + glutathione
show the reaction diagram
-
3% of the activity relative to ATP
-
-
-
UTP + gamma-Glu-L-Cys + Gly
UDP + phosphate + glutathione
show the reaction diagram
-
24% of the activity relative to ATP
-
-
-
UTP + gamma-Glu-L-Cys + Gly
UDP + phosphate + glutathione
show the reaction diagram
-
10% of the activity relative to ATP
-
-
-
ITP + gamma-Glu-L-Cys + Gly
IDP + phosphate + glutathione
show the reaction diagram
-
6% of the activity relative to ATP
-
-
-
additional information
?
-
-
the loopless mutant shows gamma-Glu-L-Cys-dependent ATP hydrolase activity to almost the same extent as its glutathione synthetase activity
-
-
-
additional information
?
-
-
the enzyme catalyzes ADP-ATP exchange at 0.62% of the rate of tripeptide synthesis
-
-
-
additional information
?
-
-
dysregulation of GSH synthesis is increasingly being recognized as contributing to the pathogenesis of many pathological conditions including diabetes mellitus, pulmonary fibrosis, cholestatic liver injury, endotoxemia and drug-resistant tumor cells
-
-
-
additional information
?
-
-
dysregulation of GSH synthesis occurs in aging and is increasingly being recognized as contributing to the pathogenesis of many pathological conditions including diabetes mellitus, pulmonary fibrosis, cholestatic liver injury, endotoxemia and drug-resistant tumor cells, detailed overview. Decrease in the activity of GS alone without a change in glutathione cysteine ligase, GCL EC 6.3.2.2, and a fall in muscle GSH levels occur after surgical trauma in human skeletal muscle
-
-
-
additional information
?
-
-
glutathione depletion is critical in cardiac dysfunction, antioxidant therapy shows positive effects on the redox state and in the disease, e.g. in metallothionein overexpressing transgenic mice or by application of buthionine sulfoximine, overview
-
-
-
additional information
?
-
-
the anti-oxidant response element promotes the expression of protective proteins including those required for glutathione synthesis, i.e. the xCT cystine antiporter, gamma-glutamylcysteine synthetase and glutathione synthase, overview
-
-
-
additional information
?
-
-
the consequences of GSH depletion include increased oxidative damage to proteins, lipids, and DNA and subsequent cytotoxic effects. GSH is also an important modulator of cellular copper homeostasis and altered Cu metabolism is central to the pathology of several neurodegenerative diseases. Both neurons and fibroblasts revealed increased expression and activation of p53 after depletion of GSH
-
-
-
additional information
?
-
Q3IFA2
the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent
-
-
-
additional information
?
-
Q3IFA2
assay method development based on [gamma32P]ATP hydrolysis coupled to the GSH synthesis, overview
-
-
-
additional information
?
-
P35669
no activity with beta-alanine and L-serine by wild-type glutathione synthetase
-
-
-
additional information
?
-
Q3IFA2
the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent, assay method development based on [gamma32P]ATP hydrolysis coupled to the GSH synthesis, overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
patients with hereditary glutathione synthetase deficiency suffer from haemolytic anaemia, 5-oxoprolinuria, metabolic acidosis, recurrent bacterial infections and various degrees of central nervous system dysfunction
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
two NFE2 sites in the human GSS promoter play important roles in the basal expression of glutathione synthetase. The glutathione synthetase gene expression is also regulated by Nrf2
-
-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + gamma-glutathione
show the reaction diagram
-
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + gamma-glutathione
show the reaction diagram
Q9FV27, Q9FVA2
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + gamma-glutathione
show the reaction diagram
Q9FV26
-
-
ir
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
-
-
-
-
-
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
-
glutathione biosynthesis
-
-
-
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q94GA0
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
AF333982
-
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
ApoE-deficient mice show reduced enzyme expression and activity, genetic regulation mechanism, overview
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
Q08220
final step in glutathione biosynthesis
-
ir
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
final step of glutathione biosynthesis
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
gene is regulated by stresses via the Atf1-Spc-1-Wis1 signal pathway
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
P48637
second step in the biosynthesis of glutahione
-
?
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
second step in the biosynthesis of glutahione
-
r
ATP + gamma-L-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
-
enzyme catalyzes the final step of glutathione biosynthesis
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P73493
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
Q3IFA2
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P35669
-
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
an optimally functioning two-step glutathione biosynthetic pathway is required in vivo for a robust defense against arsenite
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
glutathione is a ubiquitous intracellular peptide with diverse functions that include detoxification, antioxidant defense, maintenance of thiol status, and modulation of cell proliferation, whose biosynthesis is tightly regulated, overview. GSH synthase is regulated in a coordinated manner and its up-regulation can further enhance the capacity of the cell to synthesize GSH, enzyme regulation, detailed overview
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
glutathione is a ubiquitous intracellular peptide with diverse functions that include detoxification, antioxidant defense, maintenance of thiol status, and modulation of cell proliferation, whose biosynthesis is tightly regulated, overview. GSH synthase is regulated in a coordinated manner and its up-regulation can further enhance the capacity of the cell to synthesize GSH, enzyme regulation, detailed overview. Changes in GSH homeostasis occur in mice with liver-specific retinoid X receptor RXRalpha deletion
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P46416
GSH homeostasis in plants is essential for cellular redox control and efficient responses to abiotic and biotic stress. Compartmentation of the GSH biosynthetic pathway is a unique feature of plants, restricting glutathione biosynthesis to the cytosol is sufficient for normal plant development, overview
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
P46413
the enzyme catalyzes the last step in glutathione biosynthesis
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
-
the enzyme catalyzes the last step in glutathione biosynthesis, loss of intracellular neuronal GSH is an important feature of neurodegenerative disorders including Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis, overview
-
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
ADP + phosphate + glutathione
show the reaction diagram
Q3IFA2
-
-
-
?
additional information
?
-
-
dysregulation of GSH synthesis is increasingly being recognized as contributing to the pathogenesis of many pathological conditions including diabetes mellitus, pulmonary fibrosis, cholestatic liver injury, endotoxemia and drug-resistant tumor cells
-
-
-
additional information
?
-
-
dysregulation of GSH synthesis occurs in aging and is increasingly being recognized as contributing to the pathogenesis of many pathological conditions including diabetes mellitus, pulmonary fibrosis, cholestatic liver injury, endotoxemia and drug-resistant tumor cells, detailed overview. Decrease in the activity of GS alone without a change in glutathione cysteine ligase, GCL EC 6.3.2.2, and a fall in muscle GSH levels occur after surgical trauma in human skeletal muscle
-
-
-
additional information
?
-
-
glutathione depletion is critical in cardiac dysfunction, antioxidant therapy shows positive effects on the redox state and in the disease, e.g. in metallothionein overexpressing transgenic mice or by application of buthionine sulfoximine, overview
-
-
-
additional information
?
-
-
the anti-oxidant response element promotes the expression of protective proteins including those required for glutathione synthesis, i.e. the xCT cystine antiporter, gamma-glutamylcysteine synthetase and glutathione synthase, overview
-
-
-
additional information
?
-
-
the consequences of GSH depletion include increased oxidative damage to proteins, lipids, and DNA and subsequent cytotoxic effects. GSH is also an important modulator of cellular copper homeostasis and altered Cu metabolism is central to the pathology of several neurodegenerative diseases. Both neurons and fibroblasts revealed increased expression and activation of p53 after depletion of GSH
-
-
-
additional information
?
-
Q3IFA2
the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
acetate
-
anions that favor activity in decreasing order: acetate, HCOO-, Cl-, at 50 mM K+
Cd2+
-
transgenic plants overexpressing the enzyme from Escherichia coli show enhanced tolerance against cadmium, 3fold in the shoot
Cl-
-
anions that favor activity in decreasing order: acetate, HCOO-, Cl-, at 50 mM K+
Co2+
-
divalent metal ion required, Mg2+ is most effective, Co2+ can replace Mg2+
Co2+
-
77% of the efficiency relative to Mg2+
HCOO-
-
anions that favor activity in decreasing order: acetate, HCOO-, Cl-, at 50 mM K+
K+
-
5fold stimulation at 40 mM
K+
-
no requirement, but stimulates at 8-10 mM
K+
-
strict requirement for monovalent cation, maximal activity with K+
K+
-
no requirement for exchange reaction; strict requirement for monovalent cation, maximal activity with K+
K+
-
maximal activation with 50 mM; strict requirement for monovalent cation, maximal activity with K+
Li+
-
can replace K+
Li+
-
can replace K+
Mg2+
-
activates
Mg2+
-
essential for enzyme activity, optimal concentration: 10-30 mM
Mg2+
-
divalent metal ion required, Mg2+ is most effective
Mg2+
-
divalent metal ion required, Mg2+ is most effective
Mg2+
-
maximal activity at 20 mM
Mg2+
-
2 ions bound per enzyme molecule, binding structure and kinetics
Mg2+
Q08220
required, binding structure
Mg2+
-
required
Mg2+
-
-
Mg2+
-
required
Mg2+
P73493
required
Mg2+
Q3IFA2
a divalent cation is absolutely required for the activity, whereas monovalent cations are dispensable. Mg2+ is significantly more effective than Mn2+
Mg2+
P35669
required
Mn2+
-
40% of the activation rate of Mg2+, in the optimal range: 2-4 mM
Mn2+
-
can replace Mg2+
Mn2+
-
can replace Mg2+
Mn2+
-
73% of the efficiency relative to Mg2+
Mn2+
-
can partially replace Mg2+ in activation
Mn2+
-
required
NH4+
-
can replace K+
NH4+
-
can replace K+
Rb+
-
can replace K+
Zn2+
-
can partially replace Mg2+
Mn2+
Q3IFA2
a divalent cation is absolutely required for the activity, whereas monovalent cations are dispensable. Mg2+ is significantly more effective than Mn2+
additional information
-
transgenic plants overexpressing the enzyme from Escherichia coli show enhanced salt tolerance
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5,5'-dithiobis(2-nitrobenzoate)
-
inhibition of ADP-ATP exchange reaction, no effect on tripeptide synthesis
5,5'-dithiobis(2-nitrobenzoate)
-
-
5,5'-dithiobis(2-nitrobenzoate)
-
-
5,5'-dithiobis(2-nitrobenzoate)
-
Cys289 is the only amino acid residue reactive with DTNB. Modification of Cys289 with DTNB results in complete loss of the catalytic activity
7,8-dihydrofolate
-
-
adenosine
-
-
adenosine 5'-phosphosulfate
-
slight
adenosine-5'-tetraphospho-5'-pyridoxal
-
when incubated with an adenosine-5'-polyphospho-5'-pyridoxal or pyridoxal phosphate in the presence of Mg2+ and then reduced with sodium borohydride, it is most rapidly inactivated by adenosine-5'-tetraphospho-5'-pyridoxal, addition of either ATP or gamma-glutamylcysteine protects from inactivation
-
ADP
-
strong
ADP
-
product inhibition
ADP
P73493
product inhibition, competitive versus ATP, noncompetitive versus gamma-L-glutamyl-L-cysteine and glycine
Al3+
-
40% residual activity
AMP
-
slight
ATP
-
substrate inhibition above 15 mM
ATP
-
high concentrations inhibit the wild-type enzyme, while both nicked and loopless enzyme are not inhibited
Ba2+
-
27% residual activity
buthionine sulfoximine
-
reduces the GSH level and the GSH/GSSG ratio without altering GSSG levels in hearts, livers, and kidneys
buthionine sulfoximine
-
-
c-jun
AF333982
negative regulation of enzyme expression by blocking the binding of AP-1 to the promotor
-
Ca2+
-
2 mM, strong
Ca2+
-
in presence of Mg2+
Cd2+
-
in presence of Mg2+
Cd2+
-
wild-type cells are not able to grow on cadmium chloride containing substrate, but the recombinant strain is able to survive at 1 mM cadmium chloride concentration, the wild-type can grow on 1 mM Cd2+ in presence of 20 mM GSH
Cd2+
-
complete inhibition
Chloroquine
-
0.1 mM, 24% inhibition
Co2+
-
complete inhibition
Cu2+
-
CuSO4
Cu2+
-
in presence of Mg2+
Cu2+
-
15% residual activity
Fe2+
-
2 mM, strong
Fe3+
-
24% residual activity
gamma-(alpha-aminomethyl)Glu-2-aminobutyrate
-
-
gamma-Glu-Cys
-
above 2 mM
glutathione
-
product inhibition
glutathione
-
noncompetitive
glutathione
P73493
product inhibition, noncompetitive versus ATP, gamma-L-glutamyl-L-cysteine, and glycine
GSSG
-
no inhibition by reduced glutathione
GSSG
-
no inhibition by reduced glutathione
GSSG
Q3IFA2
the oxidised form of glutathione acts as an irreversible inhibitor of recombinant GshB
Hg2+
-
wild-type cells are not able to grow on mercuric chloride containing substrate, but the recombinant strain is able to survive at 0.01 mM mercuric chloride concentration, the wild-type can grow on 0.01 mM Hg2+ in presence of 20 mM GSH
Hg2+
-
complete inhibition
iodoacetate
-
-
iodoacetate
-
-
iodoacetate
-
-
L-buthionine-(S,R)-sulfoximine
-
induction of enzyme expression at 0.01 mM
L-cystine
-
1 mM, 11% inhibition
mefloquine
-
0.1 mM, 21% inhibition
menadione
-
wild-type cells are not able to grow on menadione containing substrate, but the recombinant strain is able to survive at 0.1 mM menadione, induction of enzyme expression by superoxide generation
NADPH
-
weak
NEM
-
inhibition of ADP-ATP exchange reaction, no effect on tripeptide synthesis
NEM
-
-
NEM
-
IC50: 9 mM, almost complete inhibition at 20 mM
Ni2+
-
complete inhibition
p-hydroxymercuribenzoate
-
-
PCMB
-
-
phosphate
-
product inhibition
phosphate
P73493
product inhibition, noncompetitive versus ATP, gamma-L-glutamyl-L-cysteine, and glycine
tert-butylhydroquinone
AF333982
regulatory role, activator protein 1 AP-1-mediated induction of enzyme expression, 35fold increase in activity in recombinant H4IIE cells, induction of c-jun formation, which is a negative regulatory protein blocking the binding of activator protein 1 to the promotor
trimethoprim
-
-
Zn2+
-
ZnSO4
Zn2+
-
in presence of Mg2+
methotrexate
-
-
additional information
-
enzyme expression in not affected by insulin and hydrocortisone treatment or by ethanol-feeding
-
additional information
-
no inhibition by acivicin
-
additional information
-
no induction of enzyme expression by thioacetamide
-
additional information
-
not inhibited by glutathione
-
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
-
additional information
D4N891
insensitive to feedback inhibition caused by GSH even at 20 mM
-
additional information
-
no significant inhibition: amoxycillin, gentamycin, streptomycin, kanamycin, tetracycline, penicillin, artesunate, aablaquine and primaquine
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
activator protein 1
AF333982
i.e. AP-1, mediates the induction of enzyme expression by tert-butylhydroquinone, binds to the enzyme promotor, antagonisted by c-jun, a dominant negative which is also produced in enhanced levels after treatment with tert-butylhydroquinone
-
buthionine sulfoximine
-
increase in enzyme expression in hepatocytes by 2.55fold after 18 h
diethyl maleate
-
increase in enzyme expression in hepatocytes 1.6fold
tert-butylhydroquinone
-
increase in enzyme expression in hepatocytes 1.7fold
tert-butylhydroquinone
AF333982
regulatory role, activator protein 1 AP-1-mediated induction of enzyme expression, 35fold increase in activity in recombinant H4IIE cells, induction of c-jun formation, which is a negative regulatory protein blocking the binding of activatot protein 1 to the promotor
Thioacetamide
-
increases the enzyme expression and activity in Chang cells by 50% at 6.66 mM
Thioacetamide
-
increases the enzyme expression and activity in hepatocyte cell culture and in liver 5fold in vivo
monocrotaline
-
treatment of rats with monocrotaline, a pyrrolizidine alkaloid, increases the activity of GS
additional information
-
enzyme expression in not affected by insulin and hydrocortisone treatment or by ethanol-feeding, a partial hepatectomy lead to increase in enzyme expression 2.7fold after 12 h
-
additional information
-
folate and vitamin E depletion leads to 18% increased enzyme expression and activity, mice mutants lacking apolipoprotein E show even higher activation rates of 31% on minimal diet compared to the wild-type on normal diet
-
additional information
-
(-)epicatechin upregulates the anti-oxidant response element, ARE, via an phosphatidylinositol 3-kinase-dependent pathway, ARE promotes the expression of protective proteins including those required for glutathione synthesis, i.e. the xCT cystine antiporter, gamma-glutamylcysteine synthetase and glutathione synthase, in cortical astrocytes, but not in neurons, overview, cyclic AMP response element and 3'-O-methyl-epicatechin play no significant role in enzyme upregulation
-
additional information
-
oxidative stress induces the enzyme, key transcription factors include Nrf2/Nrf1 via the antioxidant response element, ARE, activator protein-1, AP-1, and nuclear factor kappa B, NFkappaB. Nrf1 and Nrf2 overexpression induces the human GS promoter activity
-
additional information
-
oxidative stress induces the enzyme, key transcription factors include Nrf2/Nrf1 via the antioxidant response element, ARE, activator protein-1, AP-1, and nuclear factor kappa B, NFkappaB. Nrf1 is required for basal expression of GS in the mouse
-
additional information
-
oxidative stress induces the enzyme, key transcription factors include Nrf2/Nrf1 via the antioxidant response element, ARE, activator protein-1, AP-1, and nuclear factor kappa B, NFkappaB. Basal expression requires AP-1 and NFkappaB
-
additional information
P46413
tienilic acid induces the glutathione synthetase and all enzymes of the glutathione biosynthesis pathway, oxidative stress response, and phase II drug metabolism as do oxidative and/or electrophilic stresses, e.g. caused by tienilic acid, regulatory effects, overview
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.012
ATP
-
wild-type, 37C, pH not specified in the publication
0.02
ATP
-
mutant enzyme Y270H
0.033
ATP
-
-
0.037
ATP
-
recombinant enzyme, pH 8.2, 37C
0.04
ATP
-
mutant enzyme D219G
0.05
ATP
-
mutant enzyme P314L; mutant enzyme Y270C
0.057
ATP
-
25C, pH 7.5
0.057
ATP
-
wild type enzyme, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.07
ATP
-
wild-type enzyme, pH 8.2, 37C
0.07
ATP
-
wild-type enzyme
0.083
ATP
-
reaction with Glu-alpha-aminobutyrate + hydroxylamine
0.083
ATP
D7P1H2
-
0.089
ATP
-
mutant enzyme Q226A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.09
ATP
-
mutant enzyme R283C
0.116
ATP
-
mutant enzyme S153A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.12
ATP
-
mutant G371V, 37C, pH not specified in the publication
0.125
ATP
-
mutant enzyme R132K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.136
ATP
P73493
pH 7.5, 25C, wild-type enzyme
0.17
ATP
-
-
0.17
ATP
Q3IFA2
pH 7.8, 10C, recombinant enzyme, ATPase activity
0.171
ATP
-
mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.175
ATP
-
mutant enzyme R274K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.18
ATP
-
mutant enzyme E220A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.18
ATP
Q3IFA2
pH 7.8, 20C, recombinant enzyme, ATPase activity; pH 7.8, 30C, recombinant enzyme, ATPase activity
0.189
ATP
-
mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.19
ATP
-
mutant enzyme E220Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.194
ATP
-
mutant enzyme Q226N, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.2 - 1
ATP
Q3IFA2
pH 7.8, 28C, recombinant enzyme, ATPase activity
0.24
ATP
-
gamma-Glu-Cys, wild-type enzyme
0.248
ATP
-
pH 8.2, 37C
0.25
ATP
-
-
0.26
ATP
Q3IFA2
pH 7.8, 15C, recombinant enzyme, ATPase activity; pH 7.8, 30C, recombinant enzyme, with gamma-L-glutamyl-L-cysteine and glycine
0.28
ATP
Q3IFA2
pH 7.8, 25C, recombinant enzyme, ATPase activity
0.33
ATP
-
pH 8.2, temperature not specified in the publication
0.346
ATP
-
mutant enzyme E371Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.37
ATP
-
mutant enzyme R454A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.42
ATP
-
inTris-HCl buffer (200 mM, pH 8.2)
0.42
ATP
-
wild type enzyme
0.44
ATP
-
mutant G369V, 37C, pH not specified in the publication
0.48
ATP
-
-
0.53
ATP
-
pH 8.2, 25C
0.6
ATP
-
mutant with a nicked loop
0.624
ATP
-
mutant enzyme K526A
0.695
ATP
-
mutant enzyme K456M, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.73
ATP
-
mutant enzyme L188P
0.785
ATP
-
mutant enzyme N376A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.83
ATP
-
mutant K305A, pH 8.2, 37C
0.95
ATP
-
37C
1.05
ATP
-
mutant enzyme D219A
1.54
ATP
-
loopless mutant
1.8
ATP
-
-
1.85
ATP
-
-
2.66
ATP
-
mutant K305E, pH 8.2, 37C
4.9
ATP
-
mutant enzyme D448A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
13.4
ATP
-
mutant enzyme K489A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
0.32
beta-Ala
Q9M426
pH 8.0, 30C
0.592
beta-Ala
Q7XB40
pH 8.0, 30C
14
beta-Ala
-
-
170
beta-Ala
Q7X9G0, Q7X9G1
pH 8.0, 30C
80
gamma-(alpha-aminomethyl)Glu-2-aminobutyrate
-
-
0.22
gamma-Glu-2-aminobutyrate
-
-
0.5
gamma-Glu-2-aminobutyrate
-
ATP
0.5
gamma-Glu-2-aminobutyrate
-
-
0.2
gamma-Glu-Cys
-
-
0.27
gamma-Glu-Cys
-
-
0.3
gamma-Glu-Cys
-
-
0.35
gamma-Glu-Cys
-
-
0.63
gamma-Glu-Cys
-
-
0.7
gamma-Glu-Cys
-
loopless mutant
1.87
gamma-Glu-Cys
-
mutant with a nicked loop
2.6
gamma-Glu-Cys
-
-
3.33
gamma-Glu-Cys
-
-
0.091
gamma-Glu-L-2-aminobutyrate
-
-
0.03
gamma-Glu-L-Cys
Q7XB40
pH 8.0, 30C, with Gly as cosubstrate
0.035
gamma-Glu-L-Cys
-
mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.039
gamma-Glu-L-Cys
-
25C, pH 7.5
0.039
gamma-Glu-L-Cys
-
wild type enzyme, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.06
gamma-Glu-L-Cys
Q9M426
pH 8.0, 30C, with beta-Ala as cosubstrate
0.1
gamma-Glu-L-Cys
Q7X9G0, Q7X9G1
pH 8.0, 30C, with Gly as cosubstrate
0.129
gamma-Glu-L-Cys
-
mutant enzyme R132K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.143
gamma-Glu-L-Cys
-
mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.146
gamma-Glu-L-Cys
-
mutant enzyme R454A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.166
gamma-Glu-L-Cys
-
mutant enzyme S153A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.18
gamma-Glu-L-Cys
Q7X9G0, Q7X9G1
pH 8.0, 30C
0.271
gamma-Glu-L-Cys
-
mutant enzyme N376A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.273
gamma-Glu-L-Cys
-
mutant enzyme E371Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.425
gamma-Glu-L-Cys
-
mutant enzyme K456M, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.954
gamma-Glu-L-Cys
-
mutant enzyme E220Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.94
gamma-Glu-L-Cys
-
mutant enzyme E220A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.34
gamma-Glu-L-Cys
-
mutant enzyme Q226N, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.6
gamma-Glu-L-Cys
-
mutant enzyme Q226A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
5.9
gamma-Glu-L-Cys
-
inTris-HCl buffer (200 mM, pH 8.2)
5.9
gamma-Glu-L-Cys
-
wild type enzyme, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
7.03
gamma-Glu-L-Cys
-
mutant enzyme R274K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
7.5
gamma-Glu-L-Cys
-
mutant enzyme D448A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA; mutant enzyme K526A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
31
gamma-Glu-L-Cys
-
mutant enzyme K489A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
81
gamma-Glu-L-Cys
-
pH 8.2, 25C
0.28
gamma-Glu-S-methylcysteine
-
-
0.063
gamma-glutamyl-alpha-aminobutyrate
-
pH 8.2, 37C
0.1
gamma-L-Glu-aminobutanoate
-
mutant G371V, 37C, pH not specified in the publication
0.32
gamma-L-Glu-aminobutanoate
-
mutant V45W, pH 8.2, 37C
0.89
gamma-L-Glu-aminobutanoate
-
mutant G369V, 37C, pH not specified in the publication
0.95
gamma-L-Glu-aminobutanoate
-
mutant V44W, pH 8.2, 37C
1.26
gamma-L-Glu-aminobutanoate
-
wild-type, 37C, pH not specified in the publication
1.42
gamma-L-Glu-aminobutanoate
-
mutant V44A/V45A, pH 8.2, 37C; wild-type, pH 8.2, 37C
1.54
gamma-L-Glu-aminobutanoate
-
mutant V45A, pH 8.2, 37C
1.6
gamma-L-Glu-aminobutanoate
-
mutant V44A, pH 8.2, 37C
0.042
gamma-L-Glu-L-alpha-aminobutyrate
-
recombinant enzyme, pH 8.2, 37C
0.34
gamma-L-Glu-L-Cys
-
mutant K305A, pH 8.2, 37C
0.4
gamma-L-Glu-L-Cys
-
mutant K305E, pH 8.2, 37C
0.66
gamma-L-Glu-L-Cys
-
wild-type enzyme, pH 8.2, 37C
5.9
gamma-L-Glu-L-Cys
-
-
8.3
gamma-L-Glu-L-Cys
-
pH 8.2, 37C
0.099
gamma-L-glutamyl-L-cysteine
P73493
pH 7.5, 25C, wild-type enzyme
0.1 - 2
gamma-L-glutamyl-L-cysteine
-
pH 8.2, 37C, D458N
0.25
gamma-L-glutamyl-L-cysteine
-
pH 8.2, 37C, D458A
0.25
gamma-L-glutamyl-L-cysteine
Q3IFA2
pH 7.8, 30C, recombinant enzyme
0.29
gamma-L-glutamyl-L-cysteine
-
pH 8.2, 37C, D458R
0.762
gamma-L-glutamyl-L-cysteine
D7P1H2
-
1.41
gamma-L-glutamyl-L-cysteine
-
pH 8.2, 37C, wild-type enzyme
0.09
Gly
-
pH 8.2, 25C
0.18
Gly
Q7XB40
pH 8.0, 30C
0.19
Gly
-
-
0.36
Gly
-
-
0.4
Gly
-
-
0.41
Gly
-
37C
0.43
Gly
-
pH 8.2, 37C
0.53
Gly
-
wild-type, 37C, pH not specified in the publication
0.8
Gly
-
-
0.8
Gly
-
hydroxylamine
0.91
Gly
-
wild-type enzyme
1.02
Gly
-
mutant enzyme R283C
1.18
Gly
-
mutant enzyme E371Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.24
Gly
-
mutant enzyme Y270H
1.43
Gly
-
mutant enzyme Y270C
1.51
Gly
-
mutant enzyme D219G
1.51
Gly
-
25C, pH 7.5
1.51
Gly
-
wild type enzyme, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.64
Gly
-
mutant enzyme P314L
1.67
Gly
-
mutant enzyme E220Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.69
Gly
-
mutant enzyme Q226N, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.75
Gly
-
wild-type enzyme
2.07
Gly
-
mutant enzyme D219A
2.16
Gly
-
mutant enzyme R274K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
2.25
Gly
-
mutant enzyme L188P
2.25
Gly
-
mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
2.4
Gly
-
mutant enzyme K456M, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
2.52
Gly
-
mutant G371V, 37C, pH not specified in the publication
3.41
Gly
-
mutant enzyme S153A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.45
Gly
-
mutant enzyme Q226A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.53
Gly
-
-
4.04
Gly
-
mutant enzyme E220A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
4.17
Gly
-
mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
6.3
Gly
-
inTris-HCl buffer (200 mM, pH 8.2)
6.3
Gly
-
wild type enzyme, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
6.59
Gly
-
mutant enzyme N376A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
7.57
Gly
-
mutant enzyme R132K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
11.8
Gly
-
mutant enzyme K526A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
16.1
Gly
-
mutant enzyme R454A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
23.3
Gly
-
mutant with a nicked loop
23.8
Gly
-
mutant enzyme D448A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
28
Gly
-
mutant enzyme K489A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA
29.8
Gly
-
loopless mutant
0.07
glycine
Q7X9G0, Q7X9G1
pH 8.0, 30C
0.16
glycine
Q7X9G0, Q7X9G1
pH 8.0, 30C
0.25
glycine
-
mutant K305A, pH 8.2, 37C
0.407
glycine
P73493
pH 7.5, 25C, wild-type enzyme
0.452
glycine
-
pH 8.2, 37C
0.58
glycine
-
pH 8.2, 37C, wild-type enzyme
0.75
glycine
Q3IFA2
pH 7.8, 30C, recombinant enzyme
0.913
glycine
-
recombinant enzyme, pH 8.2, 37C
1.75
glycine
-
wild-type enzyme, pH 8.2, 37C
1.96
glycine
-
mutant K305E, pH 8.2, 37C
3 - 5
glycine
-
pH 8.2, 37C, D458A
4.33
glycine
D7P1H2
-
16
glycine
-
pH 8.2, 37C, D458N
19
glycine
Q9M426
pH 8.0, 30C
60
N-acetyl-L-2-aminobutyrate
-
-
25
N-acetyl-L-Cys
-
-
51
glycine
-
pH 8.2, 37C, D458R
additional information
additional information
-
-
-
additional information
additional information
-
Km-values of wild-type and mutant enzymes
-
additional information
additional information
-
Km-values for Gly and gamma-Glu-Cys are sensitive to the mutation and drastically increased
-
additional information
additional information
-
loopless mutant with increased Km-values, especially for Gly
-
additional information
additional information
-
kinetic measurements
-
additional information
additional information
-
kinetics
-
additional information
additional information
-
kinetics of cooperative substrate binding
-
additional information
additional information
-
kinetics, substrate and ligand binding, wild-type enzyme and mutants
-
additional information
additional information
-
-
-
additional information
additional information
-
the KM-value for gamma-L-Glu-L-Cys gives a distinctly nonlinear double-reciprocal plot
-
additional information
additional information
P73493
steady-state kinetics and kinetic mechanism, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.071
ATP
-
mutant enzyme R132K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.079
ATP
-
mutant enzyme R454A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.15
ATP
-
mutant enzyme K456M, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.17
ATP
-
mutant enzyme E371Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.45
ATP
-
mutant enzyme R274K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.61
ATP
-
mutant enzyme N376A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.72
ATP
-
mutant enzyme Q226A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.87
ATP
Q3IFA2
pH 7.8, 10C, recombinant enzyme, ATPase activity
1.46
ATP
-
mutant enzyme E220A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.593
ATP
-
mutant enzyme Q226N, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.85
ATP
Q3IFA2
pH 7.8, 15C, recombinant enzyme, ATPase activity
2.183
ATP
-
mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
2.42
ATP
Q3IFA2
pH 7.8, 20C, recombinant enzyme, ATPase activity
2.94
ATP
-
mutant enzyme Q226N, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase; mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3 - 6
ATP
-
mutant enzyme E220Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.183
ATP
-
mutant enzyme E220Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.3
ATP
-
mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
4.72
ATP
Q3IFA2
pH 7.8, 25C, recombinant enzyme, ATPase activity
4.99
ATP
Q3IFA2
pH 7.8, 28C, recombinant enzyme, ATPase activity
5.083
ATP
-
mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
5.32
ATP
Q3IFA2
pH 7.8, 30C, recombinant enzyme, ATPase activity
5.4
ATP
P73493
pH 7.5, 25C, wild-type enzyme
5.7
ATP
-
mutant enzyme S153A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
12.1
ATP
-
25C, pH 7.5
12.1
ATP
-
wild type enzyme, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
13
ATP
-
recombinant enzyme, pH 8.2, 37C
25.5
ATP
-
pH 8.2, 25C
55
ATP
-
inTris-HCl buffer (200 mM, pH 8.2)
0.011
gamma-Glu-L-Cys
-
mutant enzyme K456M, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.059
gamma-Glu-L-Cys
-
mutant enzyme R132K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.077
gamma-Glu-L-Cys
-
mutant enzyme R454A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.158
gamma-Glu-L-Cys
-
mutant enzyme E371Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.383
gamma-Glu-L-Cys
-
mutant enzyme N376A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.842
gamma-Glu-L-Cys
-
mutant enzyme R274K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.973
gamma-Glu-L-Cys
-
mutant enzyme Q226A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.485
gamma-Glu-L-Cys
-
mutant enzyme Q226N, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.51
gamma-Glu-L-Cys
-
mutant enzyme E220A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.13
gamma-Glu-L-Cys
-
mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase; mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.25
gamma-Glu-L-Cys
-
mutant enzyme E220Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.3
gamma-Glu-L-Cys
-
mutant enzyme S153A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.517
gamma-Glu-L-Cys
-
mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.817
gamma-Glu-L-Cys
-
mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
5.317
gamma-Glu-L-Cys
-
mutant enzyme S153A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
6.08
gamma-Glu-L-Cys
-
mutant enzyme Q226A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase; mutant enzyme Q226N, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
12.2
gamma-Glu-L-Cys
-
25C, pH 7.5
12.2
gamma-Glu-L-Cys
-
wild type enzyme, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
26.9
gamma-Glu-L-Cys
-
pH 8.2, 25C
55
gamma-Glu-L-Cys
-
inTris-HCl buffer (200 mM, pH 8.2)
13.6
gamma-L-Glu-aminobutanoate
-
mutant V45W, pH 8.2, 37C
16.4
gamma-L-Glu-aminobutanoate
-
mutant V44W, pH 8.2, 37C
17.4
gamma-L-Glu-aminobutanoate
-
mutant V45A, pH 8.2, 37C
18.9
gamma-L-Glu-aminobutanoate
-
mutant V44A, pH 8.2, 37C
19.5
gamma-L-Glu-aminobutanoate
-
wild-type, pH 8.2, 37C
21.7
gamma-L-Glu-aminobutanoate
-
mutant V44A/V45A, pH 8.2, 37C
13
gamma-L-Glu-L-alpha-aminobutyrate
-
recombinant enzyme, pH 8.2, 37C
0.003
gamma-L-Glu-L-Cys
-
mutant E144K and mutant N146D, pH 8.2, 37C
0.007
gamma-L-Glu-L-Cys
-
mutant N146A, pH 8.2, 37C
0.008
gamma-L-Glu-L-Cys
-
mutant K364A, pH 8.2, 37C
0.015
gamma-L-Glu-L-Cys
-
mutant K364E, pH 8.2, 37C
0.336
gamma-L-Glu-L-Cys
-
mutant K305E, pH 8.2, 37C
0.423
gamma-L-Glu-L-Cys
-
mutant K305A, pH 8.2, 37C
6.5
gamma-L-Glu-L-Cys
-
wild-type enzyme, pH 8.2, 37C
1.17
gamma-L-glutamyl-L-cysteine
-
pH 8.2, 37C, D458R
1.6
gamma-L-glutamyl-L-cysteine
-
pH 8.2, 37C, D458A
2.31
gamma-L-glutamyl-L-cysteine
-
pH 8.2, 37C, D458N
4.62
gamma-L-glutamyl-L-cysteine
P73493
pH 7.5, 25C, wild-type enzyme
15.68
gamma-L-glutamyl-L-cysteine
-
pH 8.2, 37C, wild-type enzyme
0.052
Gly
-
mutant G370V, 37C, pH not specified in the publication
0.062
Gly
-
mutant enzyme R132K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.08
Gly
-
mutant enzyme R454A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.133
Gly
-
mutant G369V, 37C, pH not specified in the publication
0.157
Gly
-
mutant enzyme E371Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.22
Gly
-
mutant enzyme R274K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.29
Gly
-
mutant enzyme K456M, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
0.37
Gly
-
mutant enzyme Q226A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.03
Gly
-
mutant enzyme N376A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.08
Gly
-
mutant enzyme E220A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
1.33
Gly
-
mutant enzyme Q226N, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
2.36
Gly
-
mutant G371V, 37C, pH not specified in the publication
2.467
Gly
-
mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
2.583
Gly
-
mutant enzyme E220Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
2.94
Gly
-
mutant enzyme R454K, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3 - 6
Gly
-
mutant enzyme E220Q, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
3.3
Gly
-
mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
5.183
Gly
-
mutant enzyme S155A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
7.2
Gly
-
mutant enzyme S153A, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
12.6
Gly
-
25C, pH 7.5
12.6
Gly
-
wild type enzyme, at 25C in 100 mM HEPES (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 2.5 mM gamma-glutamylcysteine, 10 mM glycine, 2.5 mM disodium ATP, 2 mM sodium phosphoenolpyruvate, 0.2 mM NADH, 5 units of type III rabbit muscle pyruvate kinase, and 10 units of type II rabbit muscle lactate dehydrogenase
18.8
Gly
-
wild-type, 37C, pH not specified in the publication
36.3
Gly
-
pH 8.2, 25C
55
Gly
-
inTris-HCl buffer (200 mM, pH 8.2)
1.17
glycine
-
pH 8.2, 37C, D458R
1.6
glycine
-
pH 8.2, 37C, D458A
2.31
glycine
-
pH 8.2, 37C, D458N
4.22
glycine
P73493
pH 7.5, 25C, wild-type enzyme
13
glycine
-
recombinant enzyme, pH 8.2, 37C
15.68
glycine
-
pH 8.2, 37C, wild-type enzyme
additional information
additional information
-
turnover-numbers of wild-type and mutant enzymes
-
additional information
additional information
-
turnover number is fatally reduced in the P227V/G240V mutant
-
additional information
additional information
-
loopless mutant with a 930fold decrease in turnover number
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5.1
ATP
Q3IFA2
pH 7.8, 10C, recombinant enzyme, ATPase activity
4
7.1
ATP
Q3IFA2
pH 7.8, 15C, recombinant enzyme, ATPase activity
4
13.4
ATP
Q3IFA2
pH 7.8, 20C, recombinant enzyme, ATPase activity
4
16.9
ATP
Q3IFA2
pH 7.8, 25C, recombinant enzyme, ATPase activity
4
23.8
ATP
Q3IFA2
pH 7.8, 28C, recombinant enzyme, ATPase activity
4
29.6
ATP
Q3IFA2
pH 7.8, 30C, recombinant enzyme, ATPase activity
4
39.71
ATP
P73493
pH 7.5, 25C, wild-type enzyme
4
46.63
gamma-L-glutamyl-L-cysteine
P73493
pH 7.5, 25C, wild-type enzyme
4246
10.36
glycine
P73493
pH 7.5, 25C, wild-type enzyme
72
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.77
ADP
-
25C, pH 7.5, with glycine as varied substrate
1.12
ADP
P73493
pH 7.5, 25C, versus ATP
1.41
ADP
P73493
pH 7.5, 25C, versus gamma-L-glutamyl-L-cysteine
1.89
ADP
-
25C, pH 7.5, with ATP as varied substrate
2.19
ADP
P73493
pH 7.5, 25C, versus glycine
2.67
ADP
-
25C, pH 7.5, with gamma-Glu-L-Cys as varied substrate
16.6
glutathione
-
25C, pH 7.5, with gamma-Glu-L-Cys as varied substrate
22.7
glutathione
-
25C, pH 7.5, with glycine as varied substrate
35.4
glutathione
-
25C, pH 7.5, with ATP as varied substrate
42.8
glutathione
P73493
pH 7.5, 25C, versus gamma-L-glutamyl-L-cysteine
54
glutathione
P73493
pH 7.5, 25C, versus ATP
68.2
glutathione
P73493
pH 7.5, 25C, versus glycine
10.7
GSSG
Q3IFA2
pH 7.8, 30C, recombinant enzyme
25.1
phosphate
-
25C, pH 7.5, with ATP as varied substrate
27.3
phosphate
P73493
pH 7.5, 25C, versus gamma-L-glutamyl-L-cysteine
29.9
phosphate
P73493
pH 7.5, 25C, versus ATP
38.6
phosphate
P73493
pH 7.5, 25C, versus glycine
47.3
phosphate
-
25C, pH 7.5, with glycine as varied substrate
69.1
phosphate
-
25C, pH 7.5, with gamma-Glu-L-Cys as varied substrate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
9
NEM
-
IC50: 9 mM, almost complete inhibition at 20 mM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0006
-
crude extract and cytosolic fraction of leaves
0.0008
-
mitochondrial fraction of leaves
0.0039
-
cell line OC/CDE22, GSH depleted
0.0047
-
cell line M22
0.0064
-
cell line OC/CDE22
0.0078
-
cell line M22, GSH depleted
0.0094
Q9FV26
bacteroid isolate
0.0127
-
bacteroid isolate
0.033
-
-
3 - 8
-
about, wild-type mice on minimal diet
11
-
purified recombinant enzyme
17.2
-
purified recombinant wild-type enzyme after complete proteoltic cleavage
17.3
-
purified recombinant isolated subfragments of 24 and 36 kDa
19.3
-
purified recombinant deletion mutant lacking 15 amino acids
20.1
-
purified recombinant wild-type enzyme
21.9
-
purified recombinant enzyme mutant with eliminated cleavage site
29.95
-
pH 8.2, 37C
32
-
about, ApoE-deficient mice on normal diet
33.5
-
about, wild-type mice on normal diet
43
-
about, ApoE-deficient mice on minimal diet
additional information
-
-
additional information
-
-
additional information
-
-
additional information
Q94GA0
-
additional information
Q9FV26
-
additional information
Q9FV27, Q9FVA2
-
additional information
-
-
additional information
-
assay development, activity under different reaction conditions in rat liver
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 7.5
-
-
7 - 8.5
-
-
7
-
assay at
7.4 - 8
-
-
7.5
P73493
assay at
8
-
assay at
8
-
assay at
8.2
-
assay at
8.5 - 9
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 9.7
-
about 50% of maximal activity at pH 7.0 and 9.7
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
P73493
assay at
37
-
assay at
37
-
assay at
37
-
assay at
37
-
assay at
37
-
assay at
45
-
-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10 - 30
Q3IFA2
the recombinant enzyme displays a great temperature-dependent increase in its activity with an unusually high value of energy of activation of 75kJ/mol for a psychrophilic enzyme
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.5
Q9FV27, Q9FVA2
isozyme GSHS2, isoelectric focusing
5.6 - 5.9
Q9FV26
isozyme GSHS2 precursor, isoelectric focusing
6.5
Q9FV27, Q9FVA2
isozyme GSHS1 precursor, isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
primary cortical astrocytes
Manually annotated by BRENDA team
-
ATCC CCL-13, cell culture
Manually annotated by BRENDA team
Q8W4W2
under optimal conditions found in bundle sheath and mesophyll cells, chilling stress does not affect total leaf GSH-S transcripts
Manually annotated by BRENDA team
Pigeon
-
-
Manually annotated by BRENDA team
P46413
expression level in presence or absence of tienilic acid, overview
Manually annotated by BRENDA team
-
primary cortical neurons
Manually annotated by BRENDA team
Q9FV27, Q9FVA2
-
Manually annotated by BRENDA team
Q9FV26
bacteroid
Manually annotated by BRENDA team
-
a non-tumorigenic cell line of oval cells established from livers of rats undergoing carcinogenesis by the choline deficient/ethionine-supplemented diet for 12 weeks
Manually annotated by BRENDA team
-
light-grown
Manually annotated by BRENDA team
-
tumorigenic cell line derived from OC22 by reiterated treatment with N-methyl-N-nitro-N-nitrosoguanidine
Manually annotated by BRENDA team
additional information
Q9FV27, Q9FVA2
isozyme GSHS2 is not expressed in leaves
Manually annotated by BRENDA team
additional information
Q9FV26
isozyme GSHS2 is not expressed in leaves
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
47-69% of the total glutathione synthetase
Manually annotated by BRENDA team
Q9FV27, Q9FVA2
isozyme GSHS2
Manually annotated by BRENDA team
Q9FV26
isozyme GSHS2
Manually annotated by BRENDA team
-
of nodule bacteroid and leaf
Manually annotated by BRENDA team
Q9FV27, Q9FVA2
isozyme GSHS1
Manually annotated by BRENDA team
Q9FV26
isozyme GSHS2
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Trypanosoma brucei brucei (strain 927/4 GUTat10.1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
70000
-
bifunctional enzyme accounts for gamma-glutamylcysteine synthetase and glutathione synthetase activities, gel filtration
662339
70000
-
gel filtration
716424
74000
Q3IFA2
recombinant His-tagged enzyme, gel filtration
728286
80000
D7P1H2
SDS-PAGE
706151
85000
-
gel filtration
1185
85000
-
SDS-PAGE
671784
85860
-
calculated from amino acid sequence
662457
118000
-
gel filtration
1194
120000
-
gel filtration
1188
120000
-
gel filtration
1207, 652466
122000 - 123000
-
gel filtration; meniscus depletion method
1197
122000 - 123000
-
meniscus depletion method
1199
122000
-
gel filtration
1198
150000
P73493
recombinant enzyme, gel filtration
726912
152000
-
gel filtration
1192
191000
-
bifunctional enzyme gamma-glutamate-cysteine ligase-glutathione synthetase, gel filtration
662457
220000
-
about 220000 Da, gel filtration
662457
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 38000, SDS-PAGE
?
-
x * 52344, calculation from nucleotide sequence
?
-
x * 36000, recombinant enzyme, SDS-PAGE
?
P35669
x * 56000, about, recombinant His-tagged wild-type and mutant enzymes, SDS-PAGE
dimer
-
-
dimer
-
2 * 56000, SDS-PAGE
dimer
-
2 * 85000, SDS-PAGE
dimer
-
2 * 60000, SDS-PAGE
dimer
-
2 * 59000, SDS-PAGE
dimer
-
2 * 59000, SDS-PAGE
dimer
-
2 * 52352, calculation from nucleotide sequence
dimer
-
2 * 61000, SDS-PAGE
dimer
Q08220
homodimer, crystal structure
dimer
-
2 * 85000, gel filtration
dimer
-
2 * 88000, gel filtration
homodimer
-
one active site for each subunit
homodimer
Q3IFA2
2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 35888, sequence calculation
homodimer
-
2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 35888, sequence calculation
-
monomer
-
x * 70000, SDS-PAGE
tetramer
-
2 * 33000 (alpha) + 2 * 26000 (beta), SDS-PAGE
tetramer
P73493
4 * 38000, recombinant enzyme, SDS-PAGE
monomer
-
bifunctional enzyme accounts for gamma-glutamylcysteine synthetase and glutathione synthetase activities
additional information
-
active site residues are Glu144, Asn146, Lys305, and Lys364, interaction of these residues with the ligands is essential for enzyme activity, especially with Glu144, but they are not essential for the overall enzyme structure
additional information
-
identification of the protease cleavage site of the 56 kDa subunit, which is cleaved into a 24 kDa and a 36 kDa fragment, also on storage at -20C, the formed heterotetramer is fully active
additional information
P35669
structure comparisons and homology structure molecular modelling of the GSH2 wild-type and mutant enzymes, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
Q9FV27, Q9FVA2
isozyme GSHS1 precursor contains a signal peptide of about 20 amino acid length, cleavage site is FFSKH-/-IPST
glycoprotein
-
contains about 2% carbohydrate
glycoprotein
-
Gal-GlyNAc located at the terminal of the oligosaccharide chain, contains fucose and mannose or glucose
proteolytic modification
Q9FV26
isozyme GSHS2 precursor contains a signal peptide of about 59 amino acid length, cleavage site is NSAPL-/-AEPD
additional information
Q9FV27, Q9FVA2
isozyme GSHS2 contains no signal peptide
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
at 2.0 A resolution
-
mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region
-
structure of Escherichia coli B glutathione synthetase complexed with ADP, glutathione, and sulfate at 2.0 A resolution
P04425
the crystal structure of the loopless mutant, in which the loop is replaced by 3 Gly residues, is identical with that of the wild-type enzyme
-
under optimal catalytic condition pH 7.5
-
computational structure modeling of wild-type and mutant enzymes using crystal structure data at 2.1 A resolution, interactions of actie site residues Glu144, Asn146, Lys305, and Lys364, and ligands ADP2-, SO42-, and Mg2+
-
purified recombinant enzyme, 20 mg/ml protein by hanging-drop method, from reservoir solution containing containing 10% w/v PEG 20K, 5% v/v tacsimate and 0.1 M HEPES, pH 7.5, 2-5 days, X-ray diffraction structure determination and analysis at 3.5 A resolution. Better crystals by vapor diffusion experiments to the without-oil microbatch method, method development, mixing of 40 mg/ml protein with an equal volume of solution containing 14% w/v PEG 20000, 0.2 M HEPES pH 7.5, 10% v/v tacsimate, X-ray diffraction structure determination and analysis at 2.34 A resolution, molecular replacement
-
crystallization by vapour-diffusion hanging-drop method of the free enzyme or the enzyme liganded to substrate gamma-glutamylcysteine and non-hydrolyzable ATP-substrate-analogue AMP-PNP, 17 mg/ml enzyme in 10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM DTT, plus equal volume of reservoir solution: for crystals of free enzyme with 1.97 M ammonium sulfate, 0.1 M Tris-HCl, pH 8.0, 2% PEG 400 at 22C, or for the liganded enzyme with 3 mM AMP-PNP, 10 mM MgCl2, 3 mM gamma-glutamylcysteine against a well solution of 2.2 M ammonium sulfate, 0.1 M Tris-HCl, pH 8.0, 2% PEG 400, 5 mM TCEP, 40 mM MgCl2, at 22C, X-ray diffraction structure determination and analysis at 2.3 A and 1.8 A, respectively
Q08220
structure modeling
-
hanging drop vapour diffusion method
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 7.5
-
stable
1192
7 - 9
-
maximal stability after 24 h
1188
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
-78
-
expression of the enzyme is absent after cryopreservation using increasing concentrations of DMSO at -78C
677199
0 - 30
Q3IFA2
purified recombinant enzyme, pH 7.8, 10 min, completely stable
728286
37
-
stable below
1192
41.3
-
melting temperature, mutant V45W
714064
42.8
-
melting temperature, mutant V44A/V45A
714064
47.8
-
melting temperature, mutant V45A
714064
50
-
pH 8.3, 40% loss of activity after 1 h in 0.15 M Tris-HCl buffer
1188
50.5
Q3IFA2
purified recombinant enzyme, pH 7.8, 10 min, 50% inactivation
728286
51.1
-
melting temperature, mutant V44A
714064
57
-
melting temperature, mutant V44W
714064
58
-
5 min, stable in absence of glutathione or other thiols
1194
58
-
relatively stable in presence of 25 mM glutathione
1197
60
-
15 min, 50% loss of activity
1192
60
-
10 min, no residual activity
716424
60
Q3IFA2
purified recombinant enzyme, pH 7.8, 10 min, inactivation
728286
60.4
-
melting temperature, wild-type
714064
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
proteolysis by arginyl endopeptidase or trypsin causes a time-dependent decrease in activity. Only one peptide bond between Arg233 and Gly234 in the loop is cleaved
-
relatively stable at all stages of purification
-
several successive freezings and thawings of the cell-free extract have no effect on enzyme activity
-
the remaining activity after arginyl-endopeptidase treatment is higher in the presence of ATP and/or gamma-Glu-Cys than in their absence
-
glycerol is required for stabilization of the enzyme during freezing
-
loss of activity by exposure to Mn2+ for long periods
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
DMSO
-
expression of the enzyme is absent after cryopreservation using increasing concentrations of DMSO at -78C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C or frozen, in presence of glutathione and 5 mM DTT, stable for 1 month
-
-18C, stable for at least 1 year
-
-20C, 20% loss of activity after 2 months
-
-20C, stable for weeks
-
-80C, 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, can be stored for up to 6 months without significant loss of activity
-
-80C, in 50 mM Tris-HCl (pH 7.4) containing100 mM NaCl, up to 6 months, without significant loss of activity
-
-20C, purified enzyme, 30% glycerol, at least 2 years stable
-
stable for more than 2 months
-
4C, -20C or -80C, the isolated enzyme loses activity quickly (about 50% in 24 h), with or without glycerol
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni2+ affinity chromatography and gel filtration
-
recombinant enzyme
-
recombinant enzyme expressed in Escherichia coli, to homogeneity
-
recombinant His-tagged wild-type enzyme from Escherichia coli, to homogeneity
-
partial from brain
-
Q-Sepharose FF HR 16/10 column chromatography, butyl-Sepharose FF HR 16/10 column chromatography, HiPrep 26/10 desalting column chromatography, Source Q anion-exchange resin chromatography, and Superdex G-200 (16/60) gel filtration; recombinant enzyme
-
from Plasmodium berghei infected mouse erythrocytes
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
from kidney to homogeneity, partially from liver
-
recombinant from Escherichia coli BL21(DE3), to homogeneity
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain DH5alpha by nickel affinity chromatography, gel filtration, and Cibacron blue 3GA affinity chromatography
P35669
recombinant His-tagged wild-type and mutant enzymes from Schizosaccharomyces pombe mutant strain, to homogeneity
-
His6-gamma-GCS-GS, bifunctional enzyme accounts for gamma-glutamylcysteine synthetase and glutathione synthetase activities
-
Ni2+-NTA resin chromatography
-
Ni2+ affinity chromatography
D7P1H2
recombinant His-tagged wild-type and mutants from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
P73493
nickel affinity chromatography and gel filtration chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning and complementation of a gsh2 mutant in fission yeast
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expression in Schizosaccharomyces pombe
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wild-type and mutant enzymes, expression in Escherichia coli
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expressed in Escherichia coli strain C41(DE3)
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gene gsh2, physical mapping and genotyping, expression analysis, overview
P46416
overexpression in an Escherichia coli expression system
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expressed in Escherichia coli BL-21DELTA(DE3) cells
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GS gene, located at chromosome 16F1, mapping of an X-linked arsenite-tolerance component to subdivision 16, overview
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cloning and amplification of a gene for glutathione synthetase
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Escherichia coli B enzyme overproduced in Escherichia coli JM109
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Escherichia coli C600 cells transformed by a recombinant plasmid for the glutathione synthetase gene of Escherichia coli B
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mutants Cys122Ala, Cys195Ala, Cys222Ala and Cys289Ala show no critical loss of activity. Multiple replacement of Cys residues, however, decreases enzymatic activity to 45-26% of the activity of the wild-type enzyme
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overexpression in Brassica juncea plants via infection with Agrobacterium tumefaciens, under control of the 35S cauliflower mosaic virus promotor
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the gene for glutathione synthetase is polymerized and cloned onto the vector plasmid pBR325
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the gene for glutathione synthetase of Escherichia coli B is cloned onto vector plasmid pBR325. Escherichia coli C600 cells transformed by a recombinant plasmid pBR325
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Q9M426
cloning of the enzyme promoter and molecular mechanisms of GS transcriptional regulation, overview. Nrf1 and Nrf2 overexpression induces the human GS promoter activity. Human GS promoter contains two regions with homology to the nuclear factor erythroid 2, NFE2, motif that are required for basal activity as mutation of these sites reduces the human GS promoter activity by 66%
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expression in Escherichia coli
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expression in Escherichia coli BL21(DE3)
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expression of N-terminally His-tagged wild-type enzyme in Escherichia coli BL21(DE3)
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gene gshB, expression of His-tagged wild-type and mutants
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seven naturally occurring missense mutations (L188P, D219A, D219G, Y270C, Y270H, R283C and P314L) are expressed using a His-tagged, Escherichia coli-based expression system
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gene gshs1, construction of cDNA library, DNA and amino acid sequence determination and analysis, expression in enzyme-deficient Escherichia coli strain 830, phylogenetic analysis reveals that gshs1 and gshs2, encoding homoglutathione synthetase, are a result of gene duplication, genomic organization
Q94GA0
Nrf1 is required for basal expression of GS in the mouse
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expressed in Escherichia coli strain K12; expression in Escherichia coli
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DNA and amino acid sequence determination
Q9FV26
DNA and amino acid sequence determination
Q9FV27, Q9FVA2
gene gshB, located on chromosome I, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
Q3IFA2
gene gshB, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
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cloning of the 2.2 kb 5'-flanking region, DNA sequence determination and analysis, promotor determination, determination of positive and negative regulation regions, e.g. NF1 repressor for induction of enzyme expression by tert-butylhydroquinone, expression in H4IIE cells, ATCC CRL-1548, via transformation by recombinant adenovirus vector
AF333982
cloning of the enzyme promoter and molecular mechanisms of GS transcriptional regulation. The rat GS promoter contains functional AP-1 sites, some of which act as enhancers. It also contains a functional NF1 site that acts as a repressor, and basal expression requires AP-1 and NFkappaB, overview
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expression in Escherichia coli BL21(DE3)
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expressed in Escherichia coli
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cloning and sequencing of the large subunit
Saccharomyces pombe
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DNA sequence analysis, overexpression of gene gshs2 in an enzyme-deficient Schizosaccharomyces pombe strain, expression of wild-type enzyme, isolated enzyme subunit fragments and mutants as C-terminally His-tagged proteins, subcloning in Escherichia coli DH5alpha
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expression in Schizosaccharomyces pombe, results in 1.4fold higher glutathione content and 1.9fold ncreased enzyme activity, regulation by the Atf1-Spc-1-Wis1 signal pathway
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sequence comparison, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain DH5alpha
P35669
expressed in Escherichia coli BL-21DELTA(DE3) cells
Q8DXM9
expression in Escherichia coli
D4N891
expression in Escherichia coli, transformation into tobacco plants
D7P1H2
gene gshB, expression of His-tagged wild-type enzyme in Escherichia coli strain BL21(DE3)
P73493
;
Q7X9G0, Q7X9G1
expression in Escherichia coli rosetta
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expression in Escherichia coli
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cloning of the large subunit
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
novel alternative splicing variant
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increased expression in response to progressive drought stress
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
DELTA67-200
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deletion mutants DELTA67-71 and DELTA67-200 are inactive
DELTA67-71
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deletion mutants DELTA67-71 and DELTA67-200 are inactive
G374V
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mutant Lys367/Pro368 to Asn/Ser and mutant Gly374 to Val are inactive
K367N/P368S
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mutant Lys367/Pro368 to Asn/Ser and mutant Gly374 to Val are inactive
E148A
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inactive enzyme
E148Q
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inactive enzyme
E220A
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decreased activity
E220Q
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decreased activity
E371A
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inactive enzyme
E371Q
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decreased activity
E429A
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inactive enzyme, decreased activity
E429Q
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inactive enzyme, decreased activity
K313M
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inactive enzyme
K367M
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inactive enzyme
N150A
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inactive enzyme
N150D
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inactive enzyme
N376A
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decreased activity
Q226A
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decreased activity
Q226N
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decreased activity
R132A
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inactive enzyme
R132K
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decreased activity
R274A
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decreased activity
R274K
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decreased activity
R454A
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decreased activity
R454K
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decreased activity
S153A
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decreased activity
P227V
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mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region
P227V/G240V
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mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region
C294A
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mutant enzymes Cys294Ala and Cys409Ala retain significant residual activity. Substantial decreases in activity are detected with mutant Cys522Ala and Cys-free mutant Cys294/Cys409/Cys422 to Ala294/Ala409/Ala422
C409A
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mutant enzymes Cys294Ala and Cys409Ala retain significant residual activity. Substantial decreases in activity are detected with mutant Cys522Ala and Cys-free mutant Cys294/Cys409/Cys422 to Ala294/Ala409/Ala422
C522A
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mutant enzymes Cys294Ala and Cys409Ala retain significant residual activity. Substantial decreases in activity are detected with mutant Cys522Ala and Cys-free mutant Cys294/Cys409/Cys422 to Ala294/Ala409/Ala422
D219A
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naturally occurring missense mutation expressed using a His-tagged, Escherichia coli-based expression system, decreases Vmax to 4% of the wild-type activity
D219G
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naturally occurring missense mutation expressed using a His-tagged, Escherichia coli-based expression system, decreases Vmax to 27% of the wild-type activity. Negative cooperativity for L-gamma-glutamyl-L-alpha-aminobutyric acid is changed to positive
D458A
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site-directed mutagenesis, the mutant shows 10% activity compared to the wild-type enzyme
D458N
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site-directed mutagenesis, the mutant shows 15% activity compared to the wild-type enzyme
D458R
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site-directed mutagenesis, the mutant shows 7% activity compared to the wild-type enzyme
E144A
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site-directed mutagenesis, 0.05% activity compared to the wild-type enzyme, unaltered tertiary structure
E144K
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site-directed mutagenesis, inactive mutant, unaltered tertiary structure
G369V
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mutation in G-loop glycine triad, about 0.7% of wild-type activity. Mutation decreases ligand binding and prevent active site closure and protection
G370V
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mutation in G-loop glycine triad, about 0.3% of wild-type activity. Mutation decreases ligand binding and prevent active site closure and protection
G371V
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mutation in G-loop glycine triad, about 13% of wild-type activity
K305A
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site-directed mutagenesis, 6.5% activity compared to the wild-type enzyme, 7fold increased Km for glycine, loss of negative cooperativity, 105fold increased Km for ATP, unaltered tertiary structure
K305E
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site-directed mutagenesis, 5% activity compared to the wild-type enzyme, loss of negative cooperativity, 40fold increased Km for ATP, unaltered tertiary structure
K364A
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site-directed mutagenesis, 0.1% activity compared to the wild-type enzyme, unaltered tertiary structure
K364E
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site-directed mutagenesis, 0.2% activity compared to the wild-type enzyme, unaltered tertiary structure
N146A
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site-directed mutagenesis, 0.1% activity compared to the wild-type enzyme, unaltered tertiary structure
N146D
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site-directed mutagenesis, 0.05% activity compared to the wild-type enzyme, unaltered tertiary structure
N146K
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site-directed mutagenesis, inactive mutant, unaltered tertiary structure
P314L
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naturally occurring neutral mutation
R283C
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naturally occurring missense mutation expressed using a His-tagged, Escherichia coli-based expression system, decreases Vmax to 13% of the wild-type activity
V44A
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decrease in melting temperature, slight decrease in activity
V44A/V45A
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decrease in melting temperature, initial activity similar to wild-type
V44W
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decrease in melting temperature, 16% decrease in activity
V45A
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decrease in melting temperature, slight decrease in activity
V45W
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decrease in melting temperature, 30% decrease in activity
Y208C
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naturally occurring missense mutation expressed using a His-tagged, Escherichia coli-based expression system, decreases Vmax to 2% of the wild-type activity
A485L/T486P
P35669
site-directed mutagenesis, the mutant shows 70% reduced activity compared to the wild-type activity, and a shift of substrate specificity with increased affinity of GSH2 for Ser as a substrate, while affinity to Gly is preserved. This provides another biosynthetic pathway for hydroxymethyl glutathione, which is known to be synthesized from glutathione and Ser in a reaction catalysed by carboxypeptidase Y
I471M/C472M/A485L/T486P
P35669
site-directed mutagenesis, the mutant shows 38% reduced activity compared to the wild-type activity, and a shift of substrate specificity with 1.2fold increased affinity of GSH2 for beta-Ala and lowered affinity for Gly, which is a characteristic of the enzyme homoglutathione synthetase found in plants, EC 6.3.2.23
D448A
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low activity
H144A
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higher activity than the wild type enzyme
K485A
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very low activity
K489A
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low activity
K526A
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very low activity
K456M
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decreased activity
additional information
P46416
construction of gsh2 insertion mutants, the mutants have a seedling lethal phenotype in contrast to the embryo lethal phenotype of gamma-glutamate cysteine ligase, gsh1, EC 6.3.2.2, null mutants, overview. The mutants show hyperaccumulation of gamma-glutamylcysteine to levels 5000-fold that in the wild-type and 200fold wild-type levels of GSH, phenotype, overview. Complementation of gsh2 mutants with the cytosol-specific GSH2 give rise to phenotypically wild-type transgenic plants
S155A
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decreased activity
additional information
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construction of transgenic plants by overexpression of the Escherichia coli enzyme, encoded by gene gshII, in the cytosol of transgenic plants, the plants show 3fold increased ability in the shoot compared to the wild-type to accumulate and tolerate heavy metals, e.g. cadmium
additional information
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creation of targeted X chromosomal deficiency lines. Relative arsenite sensitivity arises when the dose of the glutathione synthetase gene expression is reduced by half. Knockdown of GS expression by RNAi in cultured S2 cells leads to enhanced arsenite sensitivity, while GS RNAi applied to intact organisms dramatically reduces the concentration of food-borne arsenite compatible with successful growth and development, phenotypes, overview
G240V
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mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region
additional information
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deletion mutant of the loop and mutant with a nicked multifunctional loop. Cleavage of the loop results in a drastic increase in activity, which is similar to the results for the loop deletion. High concentrations of ATP inhibit the wild-type enzyme, while both nicked and loopless enzyme are not inhibited
additional information
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the mutant enzymes Arg86 and Asn283 are altered in their kinetic parameters, especially the Michaelis constant of gamma-Glu-Cys
additional information
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the crystal structure of the loopless mutant, in which the loop is replaced by 3 Gly residues, is identical with that of the wild-type enzyme. Replacement of the loop increases the Km-values, especially for glycine, and a 930fold decrease in turnover number. The loopless mutant shows gamma-Glu-L-Cys-dependent ATP hydrolase activity to almost the same extent as its glutathione synthetase activity
additional information
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construction of transgenic plants by overexpression of the enzyme in the cytosol of Brassica juncea, i.e. Indian mustard, transgenic plants show 3fold increased ability in the shoot compared to the wild-type to accumulate and tolerate heavy metals, e.g. cadmium, for use in detoxification
L188P
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naturally occurring missense mutation expressed using a His-tagged, Escherichia coli-based expression system, decreases Vmax to 9% of the wild-type activity
additional information
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all Asp458 mutants display a change in cooperativity from negative cooperativity to non-cooperative. All mutants show similar stability as compared to wild-type enzyme, differential scanning calorimetry
Y270H
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naturally occurring missense mutation expressed using a His-tagged, Escherichia coli-based expression system, decreases Vmax to 6% of the wild-type activity
additional information
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ApoE-deficient mice show reduced enzyme expression and activity
additional information
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decreased hepatic GSH levels occur, which correlated with a fall in GS activity, in Tat transgenic mice. GCLC and GS are coordinately regulated but GCLM is unchanged in liver-specific retinoid X receptor alpha knockout mice
I471M/C472V
P35669
site-directed mutagenesis, the mutant shows showed much lower affinity towards Gly and 78% reduced activity compared to the wild-type activity, but no other differences
additional information
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elimination of the protease cleavage site in the 56 kDa subunit by site-directed mutagenesis results in expression of a stable and functional enzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
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overexpression of the enzyme in transgenic plants offers a promising strategy for the production of plants with superior heavy-metal phytoremediation capacity
analysis
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assay of glutathione synthetase in erythrocytes by HPLC with fluorimetric detection, useful for rapid screening of erythrocytes glutathione synthetase activity in various pathological conditions
medicine
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the enzyme is a target for therapy of disease like diabetes mellitus, pulmonary fibrosis, cholestatic liver injury, endotoxemia and drug-resistant tumor cells, since manipulation of the GSH synthetic capacity is beneficial in treatment of many of these disorders
synthesis
D4N891
at elevated concentrations of the precursor amino acids and ATP, Escherichia coli JM109 pTrc99A-gshF produces 36 mM GSH with a molar yield of 0.9 mol/mol based on added cysteine and of 0.45 mol/mol based on added ATP. When ATP is replaced with glucose, the strain produces 7 mM in 3 h. In the presence of glucose and the pmr1 mutant of Saccharomyces cerevisiae BY4742 for ATP generation, Escherichia coli JM109 pTrc99A-gshF produces 33.9 mM GSH in 12 h with a yield of 0.85 mol/mol based on added l-cysteine
synthesis
Streptococcus thermophilus SIIM B218
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at elevated concentrations of the precursor amino acids and ATP, Escherichia coli JM109 pTrc99A-gshF produces 36 mM GSH with a molar yield of 0.9 mol/mol based on added cysteine and of 0.45 mol/mol based on added ATP. When ATP is replaced with glucose, the strain produces 7 mM in 3 h. In the presence of glucose and the pmr1 mutant of Saccharomyces cerevisiae BY4742 for ATP generation, Escherichia coli JM109 pTrc99A-gshF produces 33.9 mM GSH in 12 h with a yield of 0.85 mol/mol based on added l-cysteine
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