Information on EC 1.8.1.B1 - thioredoxin glutathione reductase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.8.1.B1
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
thioredoxin glutathione reductase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
glutathione disulfide + NADPH + H+ = 2 glutathione + NADP+
show the reaction diagram
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
no activity in Caenorhabditis elegans
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Manually annotated by BRENDA team
no activity in Canis familiaris
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Manually annotated by BRENDA team
no activity in Drosophila melanogaster
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Manually annotated by BRENDA team
no activity in Sus scrofa
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-hydroxyethyl disulfide + NADPH + H+
?
show the reaction diagram
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-
-
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?
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH
2-nitro-5-thiobenzoate + NADP+
show the reaction diagram
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-
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?
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH
?
show the reaction diagram
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-
-
-
?
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH + H+
2-nitro-5-thiobenzoate + NADP+
show the reaction diagram
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH + H+
?
show the reaction diagram
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-
-
-
?
glutaredoxin + NADPH + H+
?
show the reaction diagram
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-
-
-
?
glutathione disulfide + NADPH + H+
glutathione + NADP+
show the reaction diagram
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-
-
-
?
GSH + NADP+
GSSG + NADPH + H+
show the reaction diagram
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-
-
-
?
GSSG + NADPH + H+
glutathione + NADP+
show the reaction diagram
GSSG + NADPH + H+
GSH + NADP+
show the reaction diagram
hydroxyethyl disulfide + NADPH + H+
?
show the reaction diagram
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-
-
-
?
thioredoxin + NADP+
thioredoxin disulfide + NADPH
show the reaction diagram
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-
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?
thioredoxin + NADP+
thioredoxin disulfide + NADPH + H+
show the reaction diagram
thioredoxin disulfide + NADPH
thioredoxin + NADP+
show the reaction diagram
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absolute specificity for NADPH as electron donor
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-
?
thioredoxin disulfide + NADPH + H+
thioredoxin + NADP+
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
GSH + NADP+
GSSG + NADPH + H+
show the reaction diagram
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?
GSSG + NADPH + H+
GSH + NADP+
show the reaction diagram
thioredoxin + NADP+
thioredoxin disulfide + NADPH + H+
show the reaction diagram
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?
thioredoxin disulfide + NADPH + H+
thioredoxin + NADP+
show the reaction diagram
additional information
?
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the parasite-specific enzyme thioredoxin glutathione reductase is a component of the defense system
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
selenium
additional information
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selenoprotein
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2-oxido-1,2,5-oxadiazole-3,4-diyl)bis(2-furylmethanone)
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(2-oxido-1,2,5-oxadiazole-3,4-diyl)bis(2-thienylmethanone)
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(2-oxido-1,2,5-oxadiazole-3,4-diyl)bis[(1,3-dimethyl-1H-pyrazol-4-yl)methanone]
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(2-oxido-1,2,5-oxadiazole-3,4-diyl)bis[(4-methoxyphenyl)methanone]
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2-(4-chlorophenoxy)-6-methyl-2,3-dihydro-1,3,2-diazaphosphinin-4(1H)-one 2-oxide
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2-(4-ethoxyphenoxy)-6-methyl-2,3-dihydro-1,3,2-diazaphosphinin-4(1H)-one 2-oxide
-
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3-bromo-5-butoxy-6H-anthra[1,9-cd]isoxazol-6-one
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3-DAP
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0.05 mM, complete inhibition
4-phenyl-1,2,5-oxadiazole-3-carbonitrile 2-oxide
-
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6-methyl-2-phenoxy-2,3-dihydro-1,3,2-diazaphosphinin-4(1H)-one 2-oxide
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auranofin
aurothioglucose
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0.05 mM, complete inhibition
aurothiomalate
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0.05 mM, complete inhibition
CDE16-2
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0.05 mM, 98% inhibition
CDE4
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0.05 mM, 99% inhibition
diethyl (3-bromo-6-oxo-6H-anthra[1,9-cd]isoxazol-5-yl)propanedioate
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dimethyl (3-bromo-6-oxo-6H-anthra[1,9-cd]isoxazol-5-yl)propanedioate
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furoxan
JD159
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0.05 mM, complete inhibition
LS826
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0.05 mM, 95% inhibition
naphthazarin
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0.05 mM, complete inhibition
NOC-7
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i.e. 1-hydroxy-2-oxo-3-(N-3-methyl-aminopropyl)-3-methyl-1-triazene, complete inhibition at 0.1 mM
OPZ
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0.05 mM, 87% inhibition
P-1,3-benzothiazol-2-yl-N-(2-fluorophenyl)-P-phenylphosphinic amide
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P-1,3-benzothiazol-2-yl-N-(5-chloropyridin-2-yl)-P-phenylphosphinic amide
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P-1,3-benzothiazol-2-yl-P-phenyl-N-1,3-thiazol-2-ylphosphinic amide
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PAT
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0.05 mM, complete inhibition
potassium antimony tartrate
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RMA35
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0.05 mM, 98% inhibition
additional information
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the recombinant plasmid DNA vaccine pVAX1/SjTGR lowers enzymatic activity in vivo and in vitro
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.792
2-hydroxyethyl disulfide
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at 25°C in 0.1 M potassium phosphate (pH 7.4)
0.0345 - 0.71
5,5'-dithiobis(2-nitrobenzoic acid)
0.034
cytosolic thioredoxin
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pH 7.0, 25°C, wild-type enzyme
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1.698
GSH
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at 25°C in 0.1 M potassium phosphate (pH 7.4)
0.013 - 0.205
GSSG
0.0095 - 0.012
mitochondial thioredoxin
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0.003 - 0.08324
NADPH
0.017
thioredoxin
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pH 7.8, 25°C
0.00087 - 0.022
thioredoxin disulfide
additional information
additional information
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kinetic properties of the Grx domain of TGR
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8.65
2-hydroxyethyl disulfide
Schistosoma japonicum
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at 25°C in 0.1 M potassium phosphate (pH 7.4)
0.17 - 118
5,5'-dithiobis(2-nitrobenzoic acid)
197
cytosolic thioredoxin
Echinococcus granulosus
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pH 7.0, 25°C, wild-type enzyme
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15.78
GSH
Schistosoma japonicum
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at 25°C in 0.1 M potassium phosphate (pH 7.4)
0.19 - 25
GSSG
6.4 - 131
mitochondial thioredoxin
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4.87 - 20.1
NADPH
19.2
thioredoxin
Taenia crassiceps
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pH 7.8, 25°C
0.47 - 9
thioredoxin disulfide
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.1
2-hydroxyethyl disulfide
Schistosoma japonicum
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at 25°C in 0.1 M potassium phosphate (pH 7.4)
6426
8.7 - 119
5,5'-dithiobis(2-nitrobenzoic acid)
221
9.3
GSH
Schistosoma japonicum
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at 25°C in 0.1 M potassium phosphate (pH 7.4)
142
22 - 108
GSSG
330
58.5 - 1360
NADPH
5
540 - 1480
thioredoxin disulfide
476
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00001
auranofin
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000025 - 0.000082
(2-oxido-1,2,5-oxadiazole-3,4-diyl)bis(2-furylmethanone)
0.00002 - 0.00051
(2-oxido-1,2,5-oxadiazole-3,4-diyl)bis(2-thienylmethanone)
0.00129 - 0.00408
(2-oxido-1,2,5-oxadiazole-3,4-diyl)bis[(1,3-dimethyl-1H-pyrazol-4-yl)methanone]
0.00006 - 0.00244
(2-oxido-1,2,5-oxadiazole-3,4-diyl)bis[(4-methoxyphenyl)methanone]
0.00055
2-(4-chlorophenoxy)-6-methyl-2,3-dihydro-1,3,2-diazaphosphinin-4(1H)-one 2-oxide
Schistosoma mansoni
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-
0.0006
2-(4-ethoxyphenoxy)-6-methyl-2,3-dihydro-1,3,2-diazaphosphinin-4(1H)-one 2-oxide
Schistosoma mansoni
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-
0.00015
3-bromo-5-butoxy-6H-anthra[1,9-cd]isoxazol-6-one
Schistosoma mansoni
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-
0.0005 - 0.007
3-DAP
0.00032 - 0.0018
4-phenyl-1,2,5-oxadiazole-3-carbonitrile 2-oxide
0.004
6-methyl-2-phenoxy-2,3-dihydro-1,3,2-diazaphosphinin-4(1H)-one 2-oxide
Schistosoma mansoni
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-
0.0000032 - 0.00001
auranofin
0.00007 - 0.003
aurothioglucose
0.00005 - 0.00009
aurothiomalate
0.001 - 0.025
CDE16-2
0.0005 - 0.007
CDE4
0.009
diethyl (3-bromo-6-oxo-6H-anthra[1,9-cd]isoxazol-5-yl)propanedioate
Schistosoma mansoni
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-
0.00008
dimethyl (3-bromo-6-oxo-6H-anthra[1,9-cd]isoxazol-5-yl)propanedioate
Schistosoma mansoni
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-
0.003 - 0.0054
furoxan
0.0005 - 0.001
JD159
0.008
LS826
Schistosoma mansoni
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pH 7.4, 25°C, substrates: 5,5'-dithiobis(2-nitrobenzoic acid) + NADPH; pH 7.4, 25°C, substrates: GSSG + NADPH
0.0025 - 0.01
naphthazarin
0.05 - 0.08
OPZ
0.012 - 0.0203
P-1,3-benzothiazol-2-yl-N-(2-fluorophenyl)-P-phenylphosphinic amide
0.000789 - 0.00154
P-1,3-benzothiazol-2-yl-N-(5-chloropyridin-2-yl)-P-phenylphosphinic amide
0.0000212 - 0.000121
P-1,3-benzothiazol-2-yl-P-phenyl-N-1,3-thiazol-2-ylphosphinic amide
0.000037 - 0.000051
PAT
0.000008 - 0.000045
potassium antimony tartrate
0.004 - 0.006
RMA35
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.19
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with GSSG as substrate, at pH 7.4 and 25°C
3.33
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with thioredoxin disulfide as substrate, at pH 7.4 and 25°C
4.7
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GSSG + NADPH
8.11
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with 5,5'-dithiobis(2-nitrobenzoic acid) as substrate, at pH 7.4 and 25°C
10.3
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5,5'-dithiobis(2-nitrobenzoic acid) + NADPH
11.7
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crude extract, using GSSG as substrate, at pH 7.8 and 25°C
12.1
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with HED as substrate, at pH 7.4 and 25°C
17.5
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thioredoxin disulfide + NADPH
35
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crude extract, using 5,5'-dithiobis(2-nitrobenzoic acid) as substrate, at pH 7.8 and 25°C
128.5
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crude extract, using hydroxyethyl disulfide as substrate, at pH 7.8 and 25°C
229
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after 31fold purification, using GSSG as substrate, at pH 7.8 and 25°C
1091
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after 31fold purification, using 5,5'-dithiobis(2-nitrobenzoic acid) as substrate, at pH 7.8 and 25°C
2243
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after 31fold purification, using hydroxyethyl disulfide as substrate, at pH 7.8 and 25°C
additional information
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1536-well based kinetic absorbance assay for thioredoxin glutathione reductase. The assay uses the catalytic reduction of DTNB by NADPH in the presence of thioredoxin glutathione reductase
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2
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for 5,5'-dithiobis(2-nitrobenzoic acid) reductase activity
7.8
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for glutathione reductase activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28
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for 5,5'-dithiobis(2-nitrobenzoic acid) reductase activity
40
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for glutathione reductase activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.3
calculated from amino acid sequence
6.4
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calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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thioredoxin glutathione reductase appears to be the major, if not the sole enzyme for these activities in adult worms, completely replacing thioredoxin reductase and glutathione reductase
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
64900
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x * 64900, calculated from amino acid sequence
64940
x * 64940, calculated from amino acid sequence
136000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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1 * 66000, gel electrophoresis under denaturing conditions
homodimer
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2 * 65000
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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the glutathione reductase activity of TGR exhibits hysteretic behavior regulated by the [GSSG]/[GSH] ratio. This behavior is associated with glutathionylation by GSSG and abolished by deglutathionylation. TGR is glutathionylated at two Cys residues: Cys88 and Cys354 after 1 min incubation with 1 mM GSSG and NADPH. Absence of NADPH did not prevent glutathionylation
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure at 2.2 A resolution of TGR, deleted in the last two residues
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
continuous freezing and thawing results in loss of the enzyme activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, purified enzyme, 15 days, remains stable
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2',5'-ADP Sepharose column chromatography and Sephadex G-25 gel filtration
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ADP-agarose column chromatography
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ammonium sulfate precipitation, Ni-NTA column chromatography, GSH-Sepharose column chromatography, and 2',5'-ADP-Sepharose column chromatography
DEAE-Sephacel gel filtration, hydroxyapatite chromatography column chromatography, and Cibacron blue chromatography column chromatography
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Ni-NTA column chromatography
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nickel affinity column chromatography
recombinant enzyme
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small amounts of the soluble enzyme are obtained by purifying it from a large quantity of cells, in which protein synthesis is induced slowly for a period of 24 h at a low temperature
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of two trans-spliced mRNA variants that encode thioredoxin glutathione reductases (mitochondrial and cytosolic selenocysteine-containing isoforms) that possess identical glutaredoxin and thioredoxin reductase domains and differ exclusively in their N-termini; cloning of two trans-spliced mRNA variants that encode thioredoxin glutathione reductases (mitochondrial and cytosolic selenocysteine-containing isoforms) that possess identical glutaredoxin and thioredoxin reductase domains and differ exclusively in their N-termini
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expressed in Escherichia coli BL21 (DE3) cells
expressed in Escherichia coli BL21(DE3) cells
expression in Escherichia coli as a six-histidine tagged protein. Recombinant His-SmTGR, which would not have a penultimate selenocysteine when expressed in Escherichia coli, has no measurable activity in the insulin reduction assay
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recombinant enzyme with a fused bacterial-type SECIS element is expressed in Escherichia coli strain BL21(DE3)
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recombinant TGR is mostly insoluble when expressed in Escherichi coli, and neither cloning of the enzyme that contain various tags nor coexpression of TGR with chaperones increases its solubility
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the expression levels in 35-day-old worms and 42-day-old female worms are approximately 7times higher than the 7- and 14-day-old worms, and over 2times higher than 21- and 28-day-old worms and 42-day-old male worms
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H464Q
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mutant enzyme has only 2% of the wild-type activity. The pH dependence of Vmax for wild-type DmTrxR has pKa values of 6.4 and 9.3 on the DmTrxR-DmTrx-2 complex, whereas H464Q DmTrxR only has an observable pKa at 6.4, indicating that the pKa at pH 9.3 is contributed mainly by His464. The pKa at pH 6.4 is assigned to Cys57 and Cys490. The thiolate on Cys490 is the nucleophile in the reduction of Trx. In contrast to wild-type DmTrxR, H464Q DmTrxR does not stabilize a thiolate-FAD charge-transfer complex in the presence of excess NADPH. The rates of steps in both the reductive and the oxidative half-reactions are markedly diminished in H464Q DmTrxR as compared to those of wild-type enzyme, indicating that His464 is involved in both half-reactions
C31S
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the mutant has negligible deglutathionylase and GR activities when compared with wild type enzyme
C34S
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both the deglutathionylase and GR activities of the mutant are only slightly lower when compared with wild type enzyme
C28A
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compared to the wild type enzyme, the mutant shows 38% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 4% activity with GSSG, and 74% activity with thioredoxin disulfide
C28A/C31A
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compared to the wild type enzyme, the mutant shows 50% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 3% activity with GSSG, and 79% activity with thioredoxin disulfide
C31A
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compared to the wild type enzyme, the mutant shows 38% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 18% activity with GSSG, and 100% activity with thioredoxin disulfide
C31S
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compared to the wild type enzyme, the mutant shows 100% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 129% activity with GSSG, and 105% activity with thioredoxin disulfide
C520A
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compared to the wild type enzyme, the mutant shows 31% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 26% activity with GSSG, and 47% activity with thioredoxin disulfide
C520A/C574A
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compared to the wild type enzyme, the mutant shows 38% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 26% activity with GSSG, and 42% activity with thioredoxin disulfide
C574A
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compared to the wild type enzyme, the mutant shows 38% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 37% activity with GSSG, and 42% activity with thioredoxin disulfide
additional information
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analysis of TGR mutants reveales that the glutaredoxin domain is required for the glutathione reductase activity but does not affect the thioredoxin reductase activity. In contrast, both glutathione reductase and thioredoxin reductase activities are dependent on the Sec-containing redox center. The activity loss caused by the Sec-to-Cys mutation can be partially compensated by a Cys-to-Sec mutation of the neighboring residue, indicating that Sec can support catalysis at this alternative position
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine