BRENDA - Enzyme Database show

Arsenate reductase of Staphylococcus aureus plasmid pI258

Ji, G.; Garber, E.A.E.; Armes, L.G.; Chen, C.M.; Fuchs, J.A.; Silver, S.; Biochemistry 33, 7294-7299 (1994)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.20.4.4
overproduced in Escherichia coli
Staphylococcus aureus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.20.4.4
antimonite
SbO2-, pH 7.5, 37°C
Staphylococcus aureus
1.20.4.4
arsenite
pH 7.5, 37°C
Staphylococcus aureus
1.20.4.4
additional information
arsenite, tellurite, and antimonite [Sb(III)] are inhibitors for NADPH oxidation; the substrate, arsenate, is not inhibitory at concentrations up to 40 mM
Staphylococcus aureus
1.20.4.4
tellurite
pH 7.5, 37°C
Staphylococcus aureus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.20.4.4
additional information
-
arsenate
pH 7.5, 37°C, at low substrate concentrations the Km-value for arsenate is 0.0008 mM. Above 1 mM arsenate, a second increase in rate with increasing substrate is observed, with an apparent Km of 2 mM arsenate
Staphylococcus aureus
1.20.4.4
additional information
-
additional information
NADPH oxidation shows Michaelis-Menten kinetics with a Km of 1 microM AsO43- and an apparent Vmax of 200 nmol/min per mg of protein. At high substrate concentration (above 1 mM AsO43-), a secondary rise in the reaction rate is observed, with a Km of 2 mM and an apparent Vmax of 450 nmol/min per mg of protein
Staphylococcus aureus
1.20.4.4
0.0008
-
arsenate
high affinity
Staphylococcus aureus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.20.4.4
14436
-
x * 14436, protein with a loss of the first three amino acid residues from part of the arsenate reductase may have occurred intracellularly or extracellularly during the purification process, mass spectral analysis
Staphylococcus aureus
1.20.4.4
14440
-
calculated from amino acid sequence
Staphylococcus aureus
1.20.4.4
14500
-
electrospray mass spectrometry shows two molecular masses of 14810.5 and 14436.0 Da, suggesting that 70% of the purified protein lacks the N-terminal three amino acids
Staphylococcus aureus
1.20.4.4
14810
-
x * 14810, full length enzyme, mass spectral analysis
Staphylococcus aureus
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.20.4.4
arsenate + thioredoxin
Staphylococcus aureus
-
arsenite + thioredoxin disulfide + H2O
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.20.4.4
Staphylococcus aureus
P0A006
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.20.4.4
; gel filtration, SDS-PAGE
Staphylococcus aureus
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.20.4.4
0.2
-
pH 7.5, 37°C
Staphylococcus aureus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.20.4.4
arsenate + thioredoxin
-
639368
Staphylococcus aureus
arsenite + thioredoxin disulfide + H2O
-
-
-
?
1.20.4.4
arsenate + thioredoxin
glutaredoxin and reduced glutathione does not stimulate arsenate reduction
639368
Staphylococcus aureus
arsenite + thioredoxin disulfide + H2O
-
-
-
?
1.20.4.4
additional information
selenate is a poor substrate
639368
Staphylococcus aureus
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.20.4.4
?
x * 14436, protein with a loss of the first three amino acid residues from part of the arsenate reductase may have occurred intracellularly or extracellularly during the purification process, mass spectral analysis; x * 14810, full length enzyme, mass spectral analysis
Staphylococcus aureus
1.20.4.4
monomer
-
Staphylococcus aureus
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.20.4.4
37
-
assay at
Staphylococcus aureus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.20.4.4
7.5
-
assay at
Staphylococcus aureus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.20.4.4
additional information
purified enzyme reduces radioactive arsenate to arsenite when coupled to thioredoxin, thioredoxin reductase, and NADPH. All three protein components, arsenate reductase, thioredoxin, and thioredoxin reductase, are required for arsenate reduction. Glutaredoxin and reduced glutathione do not stimulate arsenate reduction
Staphylococcus aureus
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.20.4.4
0.0005
-
tellurite
pH 7.5, 37°C
Staphylococcus aureus
1.20.4.4
0.01
-
antimonite
pH 7.5, 37°C
Staphylococcus aureus
1.20.4.4
0.5
-
arsenite
pH 7.5, 37°C
Staphylococcus aureus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.20.4.4
overproduced in Escherichia coli
Staphylococcus aureus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.20.4.4
additional information
purified enzyme reduces radioactive arsenate to arsenite when coupled to thioredoxin, thioredoxin reductase, and NADPH. All three protein components, arsenate reductase, thioredoxin, and thioredoxin reductase, are required for arsenate reduction. Glutaredoxin and reduced glutathione do not stimulate arsenate reduction
Staphylococcus aureus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.20.4.4
antimonite
SbO2-, pH 7.5, 37°C
Staphylococcus aureus
1.20.4.4
arsenite
pH 7.5, 37°C
Staphylococcus aureus
1.20.4.4
additional information
arsenite, tellurite, and antimonite [Sb(III)] are inhibitors for NADPH oxidation; the substrate, arsenate, is not inhibitory at concentrations up to 40 mM
Staphylococcus aureus
1.20.4.4
tellurite
pH 7.5, 37°C
Staphylococcus aureus
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.20.4.4
0.0005
-
tellurite
pH 7.5, 37°C
Staphylococcus aureus
1.20.4.4
0.01
-
antimonite
pH 7.5, 37°C
Staphylococcus aureus
1.20.4.4
0.5
-
arsenite
pH 7.5, 37°C
Staphylococcus aureus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.20.4.4
additional information
-
arsenate
pH 7.5, 37°C, at low substrate concentrations the Km-value for arsenate is 0.0008 mM. Above 1 mM arsenate, a second increase in rate with increasing substrate is observed, with an apparent Km of 2 mM arsenate
Staphylococcus aureus
1.20.4.4
additional information
-
additional information
NADPH oxidation shows Michaelis-Menten kinetics with a Km of 1 microM AsO43- and an apparent Vmax of 200 nmol/min per mg of protein. At high substrate concentration (above 1 mM AsO43-), a secondary rise in the reaction rate is observed, with a Km of 2 mM and an apparent Vmax of 450 nmol/min per mg of protein
Staphylococcus aureus
1.20.4.4
0.0008
-
arsenate
high affinity
Staphylococcus aureus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.20.4.4
14436
-
x * 14436, protein with a loss of the first three amino acid residues from part of the arsenate reductase may have occurred intracellularly or extracellularly during the purification process, mass spectral analysis
Staphylococcus aureus
1.20.4.4
14440
-
calculated from amino acid sequence
Staphylococcus aureus
1.20.4.4
14500
-
electrospray mass spectrometry shows two molecular masses of 14810.5 and 14436.0 Da, suggesting that 70% of the purified protein lacks the N-terminal three amino acids
Staphylococcus aureus
1.20.4.4
14810
-
x * 14810, full length enzyme, mass spectral analysis
Staphylococcus aureus
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.20.4.4
arsenate + thioredoxin
Staphylococcus aureus
-
arsenite + thioredoxin disulfide + H2O
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.20.4.4
; gel filtration, SDS-PAGE
Staphylococcus aureus
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.20.4.4
0.2
-
pH 7.5, 37°C
Staphylococcus aureus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.20.4.4
arsenate + thioredoxin
-
639368
Staphylococcus aureus
arsenite + thioredoxin disulfide + H2O
-
-
-
?
1.20.4.4
arsenate + thioredoxin
glutaredoxin and reduced glutathione does not stimulate arsenate reduction
639368
Staphylococcus aureus
arsenite + thioredoxin disulfide + H2O
-
-
-
?
1.20.4.4
additional information
selenate is a poor substrate
639368
Staphylococcus aureus
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.20.4.4
?
x * 14436, protein with a loss of the first three amino acid residues from part of the arsenate reductase may have occurred intracellularly or extracellularly during the purification process, mass spectral analysis; x * 14810, full length enzyme, mass spectral analysis
Staphylococcus aureus
1.20.4.4
monomer
-
Staphylococcus aureus
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.20.4.4
37
-
assay at
Staphylococcus aureus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.20.4.4
7.5
-
assay at
Staphylococcus aureus