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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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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