Information on EC 1.8.5.1 - glutathione dehydrogenase (ascorbate)

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The enzyme appears in viruses and cellular organisms

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EC NUMBER
COMMENTARY hide
1.8.5.1
-
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RECOMMENDED NAME
GeneOntology No.
glutathione dehydrogenase (ascorbate)
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2 glutathione + dehydroascorbate = glutathione disulfide + ascorbate
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
ascorbate glutathione cycle
-
-
ascorbate recycling (cytosolic)
-
-
Ascorbate and aldarate metabolism
-
-
Glutathione metabolism
-
-
Metabolic pathways
-
-
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SYSTEMATIC NAME
IUBMB Comments
glutathione:dehydroascorbate oxidoreductase
-
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CAS REGISTRY NUMBER
COMMENTARY hide
9026-38-4
-
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
cultivar Hongyang
-
-
Manually annotated by BRENDA team
broccoli
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
P(SAGI2)-IPT modified plants
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
Lupinus luteus L. cv. Juno
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
alfalfa
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Prunus sp.
hybrids P3605 (Prunus amygdalus L. 'Garfi' x Prunus persica L. 'Nemared'), 8-9 (P. cerasifera L. 'P2980' x 'P3605'), 7-7 (Prunus cerasifera L. 'P2175' x Prunus davidiana L.) and 6-5 (Prunus cerasifera L. 'P2175' x Prunus amygdalus L. 'Garfi')
-
-
Manually annotated by BRENDA team
variant A101
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
-
isoform DHAR3 contributes, at least to some extent, to ascorbate recycling
physiological function
additional information
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 glutathione + dehydroascorbate
glutathione disulfide + ascorbate
show the reaction diagram
glutathione + dehydroascorbate
glutathione disulfide + ascorbate
show the reaction diagram
glutathione + dehydroascorbate
GSSG + ascorbate
show the reaction diagram
GSH + 1,2,3-trioxocyclopentane
GSSG + ?
show the reaction diagram
-
-
-
-
?
GSH + dehydroascorbate
GSSG + ascorbate
show the reaction diagram
GSH + isodehydroascorbate
GSSG + isoascorbate
show the reaction diagram
-
-
-
-
?
L-acetylcysteine + dehydroascorbate
N,N'-diacetyl-L-cystine + ascorbate
show the reaction diagram
-
4% of the activity with GSH
-
-
?
L-Cys + dehydroascorbate
? + ascorbate
show the reaction diagram
-
8% of the activity with GSH
-
-
?
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
2 glutathione + dehydroascorbate
glutathione disulfide + ascorbate
show the reaction diagram
E2RWY6
-
-
-
?
glutathione + dehydroascorbate
glutathione disulfide + ascorbate
show the reaction diagram
glutathione + dehydroascorbate
GSSG + ascorbate
show the reaction diagram
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
flavin
-
enzyme shows an unusual flavin peak, enzyme might form a flavin adduct
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
-
5 mM, 34% loss of activity
Cd2+
-
5 mM CdCl2, complete loss of activity
Cyanate
-
500 mM, 15% inhibition
dehydroascorbate
-
at high concentrations
Fe2+
-
0.1 mM FeSO4, 34% loss of activity
Fe3+
-
1 mM, complete inhibition
HgCl2
imidazole
-
1 mM, 14% loss of activity
iodoacetamide
-
-
iodoacetic acid
Mersalyl
-
-
Mn2+
-
5 mM MnSO4, 82% loss of activity
p-Chloromercuriphenylsulfonate
-
-
p-hydroxymercuribenzoate
-
1 mM, 70% inhibition
Sodium azide
-
1 mM, 32% loss of activity
additional information
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
dithiothreitol
-
1 mM, stimulates about 2.7fold
hydrogen peroxide
-
the activity of DHAR significantly increases from 1 d after treatment with 0.1 mM and thereafter declines gradually to the control level
methyl jasmonate
-
0.2 mM, activity increases after methyl jasmonate exposure and remains higher till the 9 days compared to non-treated roots
methyl viologen
-
the activity of DHAR significantly increases from 1 d after treatment with 0.05 mM and thereafter declines gradually to the control level
additional information
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.5
(25R)-3beta-hydroxycholest-5-en-27-oate
allelic variant D142 of GSTO2-2; allelic variant N142 of GSTO2-2
0.019 - 2.5
dehydroascorbate
0.04167 - 11.8
glutathione
0.69 - 5.2
GSH
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0047 - 21150
dehydroascorbate
0.0058 - 99830
glutathione
210 - 490
GSH
additional information
additional information
-
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11.62 - 44130
dehydroascorbate
39.34 - 26620
glutathione
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00568
pH 6.0
0.02632
pH 7.0
0.13
allelic variant A140/E155/E208 of GSTO1-1, dehydroascorbate as substrate, at 30°C
0.14
allelic variant A140/E155/K208 of GSTO1-1, dehydroascorbate as substrate, at 30°C; allelic variant D140/E155/E208 of GSTO1-1, dehydroascorbate as substrate, at 30°C
0.18
-
isoform DHAR2, at pH 6.5 and 25°C
0.21
allelic variant D140/DELTAE155/K208 of GSTO1-1, dehydroascorbate as substrate, at 30°C
0.22
-
isoform DHAR2, at pH 6.5 and 25°C
0.25
allelic variant A140/DELTAE155/E208 of GSTO1-1, dehydroascorbate as substrate, at 30°C
1.16
-
isoform DHAR1, at pH 6.5 and 25°C
1.77
-
-
3.22
-
isoform DHAR4, at pH 6.5 and 25°C
11.6
allelic variant D142 of GSTO2-2, dehydroascorbate as substrate, at 30°C
13.47
-
isoform DHAR2, at pH 6.5 and 25°C
13.8
allelic variant N142 of GSTO2-2, dehydroascorbate as substrate, at 30°C
18.45
-
isoform DHAR1, at pH 6.5 and 25°C
19.12
-
isoform DHAR3, at pH 6.5 and 25°C
26.15
-
isoform DHAR1, at pH 6.5 and 25°C
52
D2CGD4;
-
53
J9WN12;, J9WNR5;, J9WQY6;
pH not specified in the publication, temperature not specified in the publication; pH not specified in the publication, temperature not specified in the publication
350
-
rice recombinant DHAR
360
-
mitochondrial enzyme
400
-
plastid enzyme
965
recombinant enzyme, at pH 8.0, temperature not specified in the publication
additional information
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7
pH 5.0: about 25% of maximal activity, pH 7.0: about 65% of maximal activity
5.5 - 10
6 - 10
-
isoforms DHAR1 and DHAR2 retain more than 80% of their maximum enzymatic activities at pH values between 6.0 and 10.0, respectively
6 - 8
-
the activity of the enzyme at pH 6 was only 20% of that at pH 8.0
6.5 - 8
more than 50% activity between pH 6.5 and 8.0
6.5 - 8.5
-
pH 6.5: about 25% of maximal activity, pH 8.5: about 60% of maximal activity
6.5
J9WN12;, J9WNR5;, J9WQY6;
41% of maximum activity
6.5 - 8.5
J9WN12;, J9WNR5;, J9WQY6;
more than 66% of maximum activity; more than 66% of maximum activity
8.5
J9WN12;, J9WNR5;, J9WQY6;
86% of maximum activity
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
assay at, activity significantly higher at 30°C
30 - 35
J9WN12;, J9WNR5;, J9WQY6;
;
additional information
-
assay performed at room temperature
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15
J9WN12;, J9WNR5;, J9WQY6;
66% of maximum activity; 66% of maximum activity; 66% of maximum activity
20 - 70
20°C: about 60% of maximal activity, 70°C: about 35% of maximal activity
25 - 50
30 - 50
more than 80% activity between 30 and 50°C
55
J9WN12;, J9WNR5;, J9WQY6;
51% of maximum activity; 51% of maximum activity; 51% of maximum activity
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
calculated from amino acid sequence
8.9
calculated from sequence
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
perivascular
Manually annotated by BRENDA team
-
basal membrane
Manually annotated by BRENDA team
-
myelin sheet
Manually annotated by BRENDA team
-
from striatum
Manually annotated by BRENDA team
ripening, expression analysis, transcript level of DHAR is highest at the intermediate stage of fruit ripening, overview
Manually annotated by BRENDA team
the highest activity of the sesame DHAR is detected in the 4 week cultures of the hairy roots, after which its activity is rapidly decreased to approximately 80%
Manually annotated by BRENDA team
-
activity is significantly increased by paraquat treatment and with the increasing of paraquat concentration, particularly in the modified plants (chimeric gene P(SAGI2)-IPT transformed)
Manually annotated by BRENDA team
-
of striatum and cerebellum
Manually annotated by BRENDA team
-
within the myelin sheets
Manually annotated by BRENDA team
-
DHAR5 mRNA levels are upregulated in Arabidopsis roots and shoots colonized by the beneficial endophyticfungus Piriformospora indica
Manually annotated by BRENDA team
additional information
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
isoform DHAR3
Manually annotated by BRENDA team
additional information
-
two major dehydroascorbate reductases exist in spinach leaves. One form, DHAR-1, originates in chloroplast, the other, DHAR-b, occurs in a subcellular compartment other than chloroplast
-
Manually annotated by BRENDA team
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PDB
SCOP
CATH
UNIPROT
ORGANISM
Ectromelia virus (strain Moscow);
Homo sapiens;
Pennisetum americanum;
B9HM36
Populus trichocarpa;
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8643
x * 8643, calculated from sequence
11000
-
calculated from cDNA
15000
-
SDS-PAGE and electrospray ionization quadrupole-time-of-flight (ESI Q-TOF), His-tagged fusion protein
23000
-
gel filtration
24200
-
x * 24200, SDS-PAGE
24260
calculated from cDNA
28000
-
gel filtration
29000 - 40000
J9WN12;, J9WNR5;, J9WQY6;
gel filtration; gel filtration; gel filtration
29100
J9WN12;, J9WNR5;, J9WQY6;
1 * 29100, calculated, about 2900, MALDI-TOF; 1 * 29100, calculated, about 2900, MALDI-TOF
29690
molecular mass of the recombinant protein including a 6xHis tag determined by SDS-PAGE and MALDI-TOF/MS
29700
J9WN12;, J9WNR5;, J9WQY6;
1 * 29700, calculated, about 2900, MALDI-TOF
31000
-
x * 31000, SDS-PAGE
32000
-
1 * 32000, SDS-PAGE
48700
-
gel filtration
86000
-
gel filtration
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
isoform DHAr2 bound to glutathione, hanging drop vapor diffusion method, using 2.0 M ammonium sulfate, 0.1 M sodium acetate, pH 4.8
-
in complex with acetate or glycerol
-
in complex with ascorbate, hanging drop vapor diffusion method, using
-
crystal structures of isoform GSTO1 in complex with ascoric acid, to 1.7 A resolution. Ascorbic acid binds in the glutathione site, where the glutamyl moiety of GSH binds and stacks against a conserved aromatic residue, F34; crystal structures of isoform GSTO2-2, stabilized through site-directed mutagenesis of cysteine residues to serines and determined at 1.9 A resolution in the presence and absence of glutathione
native, ascorbate-bound, and glutathione-bound enzyme forms, hanging drop vapor diffusion method, using 0.15 M potassium bromide and 30% (w/v) PEG MME 2000
-
sitting drop vapor diffusion method, using 0.15 M potassium bromide, 30% (w/v) PEG MME 2000
-
modeling of structure. Protein has a typical glutathione S-transferase structure containing a smaller thioredoxin-like N-terminal domain and a larger helical C-terminal domain; modeling of structure. Protein has a typical glutathione S-transferase structure containing a smaller thioredoxin-like N-terminal domain and a larger helical C-terminal domain
J9WN12;, J9WNR5;, J9WQY6;
recombinant enzyme produced in Escherichia coli, crystallized by hanging-drop vapour-diffusion method
-
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 11
D2CGD4;
stable
687254
6.5
-
highest stability at, unstable under acidic and highly alkaline conditions
394028
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 60
-
the enzyme retains more than 80% activity between 30 and 60°C for 20 min
35 - 55
-
when preincubated at 35°C for 15 min, the enzyme activity is almost unaffected but begins to decrease when the temperature sets above 40°C. Isoform DHAR1 and DHAR2 still retain 61 and 73% of their initial activity when pre-incubated at 55°C for 15 min
40
-
the activity of DHAR significantly increases from 1 d after incubation at 40°C and thereafter declines gradually to the control level
42
-
pH 7.0-8.0, stable up to
53
at 53°C, both GSTO2-2 isoforms show a similar loss of activity
55
the recombinant PbDHAR is a thermostable enzyme, retaining 77% of its initial activity at 55°C
60
-
10 min, complete inactivation
65
-
complete loss of activity above
75
-
5 min, 90% loss of activity
100
-
the half-life of deactivation of the enzyme at 100°C is 8.5 min, and its thermal inactivation rate constant Kd is 0.0652/min
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
resistant to digestion by trypsin and chymotrypsin even at a high enzyme/substrate (w/w) ratio of 1:10
D2CGD4;
slight decrease in activity with increasing concentration of imidazole to 0.8 M
D2CGD4;
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, stable for at least 1 month
-
4°C, in presence of 2-mercaptoethanol, stable for several days
-
4°C, stable for at least 1 week
-
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
functional TcGrx is purified by Ni2+-nitrilotriacetic acid sepharose
-
Ni Sepharose column chromatography
Ni-agarose affinity chromatography
Ni-NTA column chromatography
Ni-NTA column chromatography and Superdex 200 gel filtration
-
Ni-NTA resin column chromatography
-
Ni2+-chealting NiNTA Superflow column chromatography
-
recombinant enzyme
using affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a fusion DHAR expressed in an Escherichia coli
enzyme form DHAR-a, expression in Escherichia coli
-
expressed in Arabidopsis thaliana
-
expressed in Arabidopsis thaliana ecotype Columbia
-
expressed in Escherichia coli
expressed in Escherichia coli BL21 cells
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3) cells, Arabidopsis thaliana, and Nicotiana benthamiana
-
expressed in Escherichia coli C41(DE3) cells
expressed in Escherichia coli NiCo21 (DE3) cells
-
expressed in Escherichia coli NiCo21(DE3) cells
-
expressed in Escherichia coli Rossetta pLysS cells
expressed in Nicotiana tabacum
expressed in Solanum tuberosum
-
expression in Escherichia coli
D2CGD4;
expression in Escherichia coli; expression in Escherichia coli; expression in Escherichia coli
J9WN12;, J9WNR5;, J9WQY6;
gene encoding DHAr, DNA and amino acid sequence determination and analysis, phylogenetic tree
overexpressed in Escherichia coli BL21 (DE3) strain using the pET-28a(+) expression vector
-
overexpression in transgenic Nicotiana tabacum plants
-
overexpression of cytosolic and plastidic isozymes in transgenic potato plants
-
overproduced in Escherichia coli
recombinant protein is overexpressed in Escherichia coli
the coding region is expressed as a His-tagged fusion protein in Escherichia coli
-
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
DHAR transcript and enzyme activity are significantly up-regulated in the leaves of acerola under cold and salt stress conditions
enzyme expression is up-regulated in response to freezing stress
-
the enzyme expression is down-regulated under salinity stress (250 mM NaCl)
the enzyme expression is up-regulated in response to high temperature (45°C)
under dark conditions, there is a sharp and significant decline in the total and reduced ascorbate contents, accompanied by a decrease in the level of transcripts and enzyme activities of the two genes in acerola leaves
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D70A
-
inactive
H95A
-
inactive
H97A
-
the mutant shows wild type activity
Q114H
-
the mutant shows reduced activity compared to the wild type enzyme
A140D
no alteration in specific activity compared to the wild type enzyme
E155
the deletion causes a 2-3fold increase in the specific activity with each substrate and a significant decrease in the enzyme's heat stability, it is also linked to abnormal arsenic excretion patterns
E208K
no alteration in specific activity compared to the wild type enzyme
N142D
no effect on the specific activity of the enzyme with any substrate
Y34A
complete loss of activity
K47A
-
the mutant shows about wild type activity
K8A
-
the mutation significantly reduces the enzymatic activity
D72A
J9WN12;, J9WNR5;, J9WQY6;
reduction in catalytic efficincy. D72 is a glutathione-site residue
K8A
J9WN12;, J9WNR5;, J9WQY6;
severe reduction in catalytic efficincy. K8 is a glutathione-site residue
S73A
J9WN12;, J9WNR5;, J9WQY6;
reduction in catalytic efficincy. S73 is a glutathione-site residue
C23S
-
mutant enzyme has almost no activity
C26S
-
turnover number is 57% of the wild-type value, KM-value for dehydroascorbate is 2fold lower than the wild-type value, KM-value for GSH is 1.6old lower than the wild-type value
C9S
-
turnover number is 86% of the wild-type value, KM-value for dehydroascorbate is 2.8fold lower than the wild-type value, KM-value for GSH is 2fold lower than the wild-type value
C9S/C26S
-
turnover number is 43% of the wild-type value, KM-value for dehydroascorbate is 1.1fold higher than the wild-type value, KM-value for GSH is identical to the wild-type value
C25S
-
has equivalent specificity constants for dehydroascorbate and GSH, but may have a different catalytic mechanism
additional information
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Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
GSTO2-2, using 8 mM urea
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
medicine
significant association with the age-at-onset of Alzheimer's disease and Parkinson's disease
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LINKS TO OTHER DATABASES (specific for EC-Number 1.8.5.1)
ExplorEnz
ExPASy
KEGG
MetaCyc
SABIO-RK
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
InterPro (database of protein families, domains and functional sites)