Information on EC 1.8.99.B1 - protein-disulfide reductase (acceptor)

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

EC NUMBER
COMMENTARY hide
1.8.99.B1
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
protein-disulfide reductase (acceptor)
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
protein-dithiol + oxidized acceptor = protein-disulfide + reduced acceptor
show the reaction diagram
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Q9YDZ4
UniProt
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
dithiol peptide NRCSQGSCWN + acceptor
disulfide peptide NRCSQGSCWN + reduced acceptor
show the reaction diagram
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?
insulin disulfide + reduced thioredoxin
insulin dithiol + pxodized thioredoxin
show the reaction diagram
protein-dithiol + oxidized acceptor
protein-disulfide + reduced acceptor
show the reaction diagram
[insulin]-disulfide + reduced dithiothreitol
[insulin]-dithiol + oxidized dithiothreitol
show the reaction diagram
[ribonuclease A]-dithiol + oxidized acceptor
[ribonuclease A]-disulfide + reduced acceptor
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
protein-dithiol + oxidized acceptor
protein-disulfide + reduced acceptor
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 30
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assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.7
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calculated from sequence
5.3
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calculated from sequence
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6200
x * 6200, reducing SDS-PAGE
19930
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1 * 19930, calculated from sequence
20995
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1 * 20995, light scattering and electrospray mass spectroscopy analyses
21000
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light scattering and electrospray mass spectroscopy analyses
25650
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wild-type enzyme electrospray mass spectroscopy
26032
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x * 26032, calculated from sequence
26700
gel-filtration, electrospray mass spectroscopy
27100
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gel filtration, electrospray mass spectrometry
27198
Q9YDZ4
1 * 27198, calculated from sequence, gel filtration, electrospray mass spectroscopy
27200
Q9YDZ4
gel filtration, electrospray mass spectroscopy
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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the protein consists of two homologous structural units with low sequence identity. Each unit contains a thioredoxin fold with a distinct CXXC active site motif
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystals are grown in the presence of 2 M ammonium sulfate, 2% (v/v) PEG 400, 0.1 M Hepes (pH 8). X-ray diffraction data are collected to 1.93 A resolution
Q9YDZ4
crystal structure at 2.4 A resolution
purified recombinant enzyme, hanging-drop vapor-diffusion method, mixing of 0.001 ml of 5.5 mg/ml protein solution with 0.001 ml of reservoir solution containing 13% PEG 20000, and 0.1 M MES buffer, pH 6.0, method optimization, X-ray diffraction structure determination and analysis at 1.80 A resolution, molecular modelling
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the X-ray crystallographic structure is solved at 1.80 A resolution
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 90
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stable up to 85°C, enzyme melting point, circular dichroism spectroscopy thermal denaturation study
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
guanidine-HCl
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant C-terminally His-tagged enzyme from Escherichia coli strain BL21-Codon Plus (DE3)RIL by ultracentrifugation, heat treatment at 70°C for 20 min, metal chelating affinity chromatography, dialysis, anion exchange chromatography, and again dialysis
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wild-type enzyme and recombinant mutant enzymes C34S, C148S and C34S/C148S
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
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expression in Escherichia coli
expression of wild-type and mutant enzymes C35S, C146S and C35S/C146S in Escherichia coli
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gene Sso1120, recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21-Codon Plus (DE3)RIL
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mutant enzymes C34S, C148S and C34S/C148S, are overexpressed in Escherichia coli
wild-type enzyme and mutant enzymes C155S/C158S, C41S/C44S and C173S/C178S are expressed in Escherichi coli
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
when Sulfolobus solfataricus cells are incubated with H2O2, paraquat and tert-butyl hydroperoxide, the Sso0192 mRNA level increases. Specifically, a two-fold increase in transcriptional levels is observed within 30 min of the addition of tert-butyl hydroperoxide, while a slight induction is observed 30 min after paraquat and H2O2 treatment
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C148S
mutant enzyme exhibits a lower activity compared to wild-type enzyme
C34S
mutant enzyme retains nearly wild-type activity
C34S/C148S
mutant enzyme shows no activity
C146S
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not active in the insulin reductase assay, oxidation of the dithiol peptide NRCSQGSCWN is reduced
C35S
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active in the insulin reductase assay, oxidation of the dithiol peptide NRCSQGSCWN is comparable to wild-type enzyme
C35S/C146S
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not active in the insulin reductase assay, no oxidation of the dithiol peptide NRCSQGSCWN
C155S/C158S
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with insulin as substrate the enzyme is completely inactive
C173S/C178S
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with insulin as substrate the enzyme shows lower activity than wild-type enzyme
C41S/C44S
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with insulin as substrate the enzyme shows lower activity than wild-type enzyme
C155S/C158S
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with insulin as substrate the enzyme is completely inactive
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C173S/C178S
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with insulin as substrate the enzyme shows lower activity than wild-type enzyme
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C41S/C44S
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with insulin as substrate the enzyme shows lower activity than wild-type enzyme
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