Information on EC 1.20.4.1 - arsenate reductase (glutaredoxin)

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

EC NUMBER
COMMENTARY
1.20.4.1
-
RECOMMENDED NAME
GeneOntology No.
arsenate reductase (glutaredoxin)
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
arsenate + glutaredoxin = arsenite + glutaredoxin disulfide + H2O
show the reaction diagram
-
-
-
-
arsenate + glutaredoxin = arsenite + glutaredoxin disulfide + H2O
show the reaction diagram
reaction mechanism
-
arsenate + glutaredoxin = arsenite + glutaredoxin disulfide + H2O
show the reaction diagram
arsenate reduction by glyceraldehyde-3-phosphate dehydrogenase may take place during or as a consequence of, the arsenolytic cleavage of the thioester bond formed between the enzymes C149 and the 3-phosphoglycerol moiety of the substrate
-
arsenate + glutaredoxin = arsenite + glutaredoxin disulfide + H2O
show the reaction diagram
first reaction step is a nucleophilic displacement reaction by C10 on dianionic arsenate. Second step is a preferential nucleophilic attack of C82 on the monoanionic C10-arsenate intermediate stabilized by S17. Thiolate form of C82 is stabilized by an eight-residue alpha helix flanked by C82 and C89 and a hydrogen bond with T11. during the final step, C89 is activated as a nucleophile by structural alterations of the redox helix
P0A006
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
arsenate detoxification I (glutaredoxin)
-
arsenate detoxification II (glutaredoxin)
-
conversions
BRENDA
BRENDA
BRENDA
SYSTEMATIC NAME
IUBMB Comments
glutharedoxin:arsenate oxidoreductase
A molybdoenzyme. The glutaredoxins catalyse glutathione-disulfide oxidoreductions and have a redox-active disulfide/dithiol in the active site (-Cys-Pro-Tyr-Cys-) that forms a disulfide bond in the oxidized form [2, 10]. Glutaredoxins have a binding site for glutathione, which is required to reduce them to the dithiol form [3,6]. Thioredoxins reduced by NADPH and thioredoxin reductase can act as alternative substrates. The enzyme [1, 4, 7, 9] is part of a system for detoxifying arsenate. Although the arsenite formed is more toxic than arsenate, it can be extruded from some bacteria by EC 3.6.3.16, arsenite-transporting ATPase; in other organisms, arsenite can be methylated by EC 2.1.1.137, arsenite methyltransferase, in a pathway to non-toxic organoarsenical compounds.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Acr2p
-
-
-
-
ACR3-ArsC
-
gene name, protein comprises two domains an aquaglyceroporin-derived N-terminal channel-like part fused to a C-terminal enzyme domain with similarity to ArsC arsenate reductase
All0195
Q8Z0A3
-
All0195
Nostoc sp. PCC7120
Q8Z0A3
-
-
ArsC
-
-
-
-
ArsC
P74313
-
arsenate reductase
-
-
arsenate reductase
-
-
arsenate reductase
-
-
-
arsenate reductase
-
-
arsenate reductase
-
-
EC 1.97.1.5
-
-
formerly
-
gene arsC proteins
-
-
-
-
glutaredoxin
Q1A5X4
-
glutathione:arsenate oxidoreductase
-
-
-
-
Grx
Q1A5X4
-
PcAcr2
Q1A5X4
-
proteins (specific proteins and subclasses), gene arsC
-
-
-
-
PvGRX5
Q1A5X4
-
reductase, arsenate
-
-
-
-
slr0946
-
gene name
CAS REGISTRY NUMBER
COMMENTARY
146907-46-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
glyceraldehyde-3-phosphate dehydrogenase working as arsenate reductase
-
-
Manually annotated by BRENDA team
Nostoc sp. PCC7120
-
UniProt
Manually annotated by BRENDA team
5fold increase in enzyme activity after 9 days treatment with 2 mM arsenate; Chinese brake fern, an arsenic hyperaccumulator
-
-
Manually annotated by BRENDA team
glyceraldehyde-3-phosphate dehydrogenase working as arsenate reductase
-
-
Manually annotated by BRENDA team
ArsC protein from cyanobacterium Synechocystis sp. strain PCC 6803 (SynArsC) is related to the thioredoxin-dependent ArsC family, but uses the glutathione/glutaredoxin system for arsenate reduction (thioredoxin/glutaredoxin hybrid arsenate reductase)
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
as a dual functional protein (arsenite channel and arsenate reductase) ACR3-ArsC rescues a yeast strain that is highly sensitive to arsenate due to deletion of the ACR2 reductase and all transport proteins for arsenite, ACR3, Fps1, and the vacuolar ABC transporter Ycf1 for arsenite-thiol conjugates
physiological function
Q8Z0A3
enzyme complements the arsenate-sensitive phenotype of Escherichia coli
physiological function
Nostoc sp. PCC7120
-
enzyme complements the arsenate-sensitive phenotype of Escherichia coli
-
physiological function
-
as a dual functional protein (arsenite channel and arsenate reductase) ACR3-ArsC rescues a yeast strain that is highly sensitive to arsenate due to deletion of the ACR2 reductase and all transport proteins for arsenite, ACR3, Fps1, and the vacuolar ABC transporter Ycf1 for arsenite-thiol conjugates
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
arsenate + reduced acceptor
arsenite + acceptor
show the reaction diagram
-
-
-
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
-
-
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
-
-
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
strong specificity for arsenate
-
-
-
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
thioredoxin is unable to support arsenate reduction. The N-terminal Cys residue is essential for arsenate reductase activity. During the catalytic cycle, Acr2p forms a mixed disulfide with GSH before being reduced vby glutaredoxin to regenerate the active Acr2p reductase
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the first step of the reaction is the binding of arsenate, followed by the interaction of the enzyme-arsenate complex with GSH. A reaction scheme is hypothesized in which the enzyme forms a mixed disulfide between the Cys-12 thiolate of ArsC and GSH. Glutaredoxin would then be required to resolve the mixed disulfide, regenerating reduced ArsC
-
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
thioredoxin is not effective as electron donor
-
-
-
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme uses GSH with glutaredoxin as electron donor. Glutaredoxin 2 is the most effective hydrogen donor for the reduction of arsenate. During the catalytic cycle, ArsC forms a mixed disulfide with GSH before being reduced by glutaredoxin to regenerate the active ArsC reductase
-
-
-
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
C12 is located at the active site and is required for catalysis
-
-
-
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
Q8Z0A3
enzyme catalyzes the oxidation of NADPH coupled with reduction of arsenate in the presence of glutathione reductase, glutathione and glutaredoxin
-
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
A7MRG2, -
glutaredoxin functions as the electron donor for arsenate reduction
-
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
Nostoc sp. PCC7120
Q8Z0A3
enzyme catalyzes the oxidation of NADPH coupled with reduction of arsenate in the presence of glutathione reductase, glutathione and glutaredoxin
-
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
A7MRG2, -
glutaredoxin functions as the electron donor for arsenate reduction
-
-
?
arsenate + reduced glutathione
arsenite + glutathione
show the reaction diagram
-
-
-
-
?
arsenate + reduced glutathione
arsenite + glutathione
show the reaction diagram
Q1A5X4
-
-
-
?
arsenate + reduced glutathione
arsenite + glutathione
show the reaction diagram
-
enzyme plays an important role in detoxification of arsenate
-
-
?
arsenate + reduced glutathione + NAD+ + glyceraldehyde-3-phosphate
arsenite + glutathione + ?
show the reaction diagram
-
-
-
-
?
additional information
?
-
Q1A5X4
both glutathione-SH and glutaredoxin are required for activity. No substrate: phosphate
-
-
-
additional information
?
-
-
no substrate: phosphate, nitrate
-
-
-
additional information
?
-
P74313, -
chemical shift assignments of 1H, 13C and 15N atoms for the reduced form the enzyme
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
arsenate + reduced acceptor
arsenite + acceptor
show the reaction diagram
-
-
-
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutaredoxin
arsenite + oxidized glutaredoxin
show the reaction diagram
-
the enzyme is involved in bacterial arsenic resistance
-
?
arsenate + reduced glutathione
arsenite + glutathione
show the reaction diagram
-
enzyme plays an important role in detoxification of arsenate
-
-
?
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
glutaredoxin
-
-
glutaredoxin
-
replacement of the active-site Cys15 by serine completely eliminates the ability of glutatredoxin A to serve as an electron donor. Replacement of either of the two cysteine residues distant from the active site, i.e., Cys 36 and Cys70, has no effect on the electron-donating ability of glutaredoxin
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
arsenite
-
noncompetitive
diethyldicarbonate
-
-
iodoacetate
-
inactivates wild type enzyme and mutant enzyme C106S, in presence of substrate, arsenate, or the competitive inhibitors phosphate or sulfate, the rate of the alkylation is reduced
Koningic acid
-
inhibition of both classical enzymic and arsentate reductase activities of glyceraldehyde-3-phosphate dehydrogenase
phosphate
-
poor
phosphate
-
competitive
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
arsenate
-
10 mM increases enzyme activity by 1.6fold
GSH
-
glutathione is not effective as electron donor
GSH
-
the enzyme uses GSH with glutaredoxin as electron donor
additional information
-
enzyme is arsenate-inducible
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0002
-
arsenate
-
mutant C13A, pH 7.5, temperature not specified in the publication; mutant C35A, pH 7.5, temperature not specified in the publication; wild-type, pH 7.5, temperature not specified in the publication
2.3
-
arsenate
-
pH 6.5, 30C
2.33
-
arsenate
-
pH 6.5, 30C
8
-
arsenate
-
-
28
-
arsenate
-
pH 6.5
0.000002
-
glutaredoxin
-
glutaredoxin 2 from E. coli
0.0001
-
glutaredoxin
-
glutaredoxin 3 from E. coli
0.0006
-
glutaredoxin
-
glutaredoxin 1p from Saccharomyces cerevisiae or glutaredoxin 1 from E. coli
0.0042
-
glutaredoxin 1
-
-
-
0.000003
-
glutaredoxin 2
-
-
-
0.3
-
glutaredoxin 3
-
-
-
0.57
-
reduced glutathione
-
pH 6.5, 30C
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.3
-
glutaredoxin 1
-
-
-
0.14
-
glutaredoxin 2
-
-
-
0.09
-
glutaredoxin 3
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.45
-
-
root
3.84
-
-
shoot
4.8
-
-
isolated roots in arsenate containiing culture medium, basic level
24
-
-
isolated roots in arsenate containiing culture medium, induced for 9 h by arsenate
additional information
-
-
in vivo activity after feeding experiments
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.3
6.8
-
-
6.5
-
-
; broad optimum
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
7.5
-
90% of maximal activity at pH 5.5, 60% at pH 7.5
5.5
-
-
90% of maximum activity
7.5
-
-
60% of maximum activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
50
-
broad optimum
additional information
-
-
reductase activity increases from 10C to 40C
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
50
-
; broad
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8.9
-
-
calculated
8.9
-
Q8Z0A3
calculated
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
enzyme activity is constitutive and unaffected by presence of arsenate
Manually annotated by BRENDA team
additional information
-
no activity in shoot, arsenite is the main storage form in the fern fronds, no enzymic activity in shoots
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
besides glyceraldehyde-3-phosphate dehydrogenase, presence of other enzymes contributing to arsenate reduction
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Synechocystis sp. (strain PCC 6803 / Kazusa)
Synechocystis sp. (strain PCC 6803 / Kazusa)
Synechocystis sp. (strain PCC 6803 / Kazusa)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
34000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 14400, SDS-PAGE
?
-
x * 14500, calculated and SDS-PAGE
?
Q8Z0A3
x * 13624, calculated
?
-
x * 15000, SDS-PAGE, x* 15507, MALDI-TOF, His-tagged recombinant protein
?
Nostoc sp. PCC7120
-
x * 13624, calculated
-
dimer
-
2 * 16822, calculation from nucleotide sequence
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
molecular modeing of structure. Residues Cys12, Arg60, Arg94, and Arg107 are identified as metal binding residues
A7MRG2, -
comparative homology modeling. Along with Cys11, the residues like Ile8, Pro9, Asn10, Gly12, Thr13, Cys14, Lys15, and Phe18 also show coordination with arsenate
Q8Z0A3
in silico modeling of disulfide bonds into the solution NMR structure of the fully-reduced enzyme. C8/C82 and C80C82 disulfides are equally consistent
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
affinity purified
-
using Ni-NTA chromatography, gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli complements phenotypes
-
-
Q8Z0A3
gene Pv5-6 encoding PvGrx5 expressed in Escherichia coli strain XL-1 Blue. PvGrx5 expressed in Escherichia coli mutants in which the arsenic resistance genes of the ars operon are deleted (strain AW3110), a deletion of the gene for the ArsC arsenate reductase (strain WC3110), and a strain in which the ars operon is deleted and the gene for the GlpF aquaglyceroporin is disrupted (strain OSBR1)
-
sequence contains a variation of conserved motif HCX5R where R is replaced by S
-
cloned into the bacterial expression vector and expressed in Escherichia coli as a C-terminal histidine-tagged protein
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A11W
-
same Km for arsenate as the wild type with maximal velocity approximately half that of the wild type enzyme
A11W/C12S
-
catalytic inactive mutation
C106G
-
the maximal velocity is approximately half that of the wild type enzyme
C12S
-
catalytically inactive mutant
Y7W
-
same Km and maximal velocity as the wild type
C67A
-
loss of both GRX activity and arsenic resistance
N13A
P0A006
decrease in activity due to lower local softness, analysis of S-As bond length, Wiberg bond orders, lewaving group energy and nucleofugality
R16A
P0A006
decrease in activity due to lower local softness, analysis of S-As bond length, Wiberg bond orders, lewaving group energy and nucleofugality
S17A
P0A006
decrease in activity due to lower local softness, analysis of S-As bond length, Wiberg bond orders, lewaving group energy and nucleofugality
C13A
-
114% of wild-type vmax
C35A
-
l00% of wild-type vmax
C80A
-
less than 2% of wild-type vmax
C80A/C82A
-
less than 2% of wild-type vmax
C82A
-
less than 2% of wild-type vmax
C8A
-
less than 2% of wild-type vmax
additional information
-
a disruption mutant is inactive
additional information
-
redcution of enzyme expression by RNAi to 2% of wild-type level. Knockdown plant lines are more sensitive to high concentrations of arsenate, but not arsenite and accumulate 10- to 16fold more arsenic in shoots than wild-type when grown on arsenate medium
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
degradation
-
PvGrx5 has a role in regulating intracellular arsenite levels, by either directly or indirectly modulating the aquaglyceroporin