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Literature summary extracted from

  • Messens, J.; Hayburn, G.; Desmyter, A.; Laus, G.; Wyns, L.
    The essential catalytic redox couple in arsenate reductase from Sataphylococcus aureus (1999), Biochemistry, 38, 16857-16865.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.20.4.4 expressed in Escherichia coli Staphylococcus aureus
1.20.4.4 expression in Escherichia coli. Wild-type enzyme and the Cys mutants (C15A, C10A, C82A, C82S, C89A, C10SC15S, C10SC15A) are expressed in Escherichia coli. Wild-type enzyme, mutant enzyme C15A, mutant enzyme C10A, mutant enzyme C82S, mutant enzyme C89A, and mutant enzyme C10SC15A are expressed soluble and with high yields. Mutant enzyme C82A is found in inclusion bodies, and the double mutant C10S/C15S is not expressed Staphylococcus aureus

Protein Variants

EC Number Protein Variants Comment Organism
1.20.4.4 C10A inactive mutant enzyme Staphylococcus aureus
1.20.4.4 C10A site-directed mutagenesis, mutation of Cys 10, 82, and 89 leads to redox-inactive enzymes Staphylococcus aureus
1.20.4.4 C10S/C15A double mutation, no enzymatic activity Staphylococcus aureus
1.20.4.4 C10S/C15S double mutation, no enzymatic activity Staphylococcus aureus
1.20.4.4 C10SC15A inactive mutant enzyme Staphylococcus aureus
1.20.4.4 C15A site-directed mutagenesis, only ArsC wild type and ArsC C15A show enzymatic activity Staphylococcus aureus
1.20.4.4 C15A as compared to wild-type enzyme the affinity is reduced ba a factor of 2 Staphylococcus aureus
1.20.4.4 C82A site-directed mutagenesis, mutation of Cys 10, 82, and 89 leads to redox-inactive enzymes Staphylococcus aureus
1.20.4.4 C82S inactive mutant enzyme Staphylococcus aureus
1.20.4.4 C82S site-directed mutagenesis, mutation of Cys 10, 82, and 89 leads to redox-inactive enzymes Staphylococcus aureus
1.20.4.4 C89A inactive mutant enzyme Staphylococcus aureus
1.20.4.4 C89A site-directed mutagenesis, mutation of Cys 10, 82, and 89 leads to redox-inactive enzymes Staphylococcus aureus
1.20.4.4 additional information essential cysteinyl residues and redox couple in arsenate reductase are identified by a combination of site-specific mutagenesis and endoprotease-digest mass spectroscopy analysis Staphylococcus aureus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.20.4.4 0.000066
-
 arsenate wild-type Staphylococcus aureus
1.20.4.4 0.000066
-
 arsenate pH 7.5, 37°C, wild-type enzyme Staphylococcus aureus
1.20.4.4 0.000134
-
 arsenate C15A mutant Staphylococcus aureus
1.20.4.4 0.000134
-
 arsenate pH 7.5, 37°C, mutant enzyme C15A Staphylococcus aureus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.20.4.4 14400
-
 gel filtration Staphylococcus aureus
1.20.4.4 21000
-
 gel filtration Staphylococcus aureus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.20.4.4 arsenate + thioredoxin Staphylococcus aureus the enzyme encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 reduces intracellular arsenate to the more toxic arsenite, which is subsequently extruded from the cell arsenite + thioredoxin disulfide + H2O
-
 ?
1.20.4.4 additional information Staphylococcus aureus assays are performed with different arsenate concentrations and arsenate reductase concentrations in the presence of 0.42 microM Escherichia coli thioredoxin, 0.14 microM Escherichia coli thioredoxin reductase and 125 microM NADPH ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.20.4.4 Staphylococcus aureus P0A006
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.20.4.4
-
 Staphylococcus aureus
1.20.4.4 wild-type ArsC and the Cys mutants, gel filtration more than 97% pure Staphylococcus aureus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.20.4.4 arsenate + thioredoxin
-
 Staphylococcus aureus arsenite + thioredoxin disulfide + H2O
-
 ?
1.20.4.4 arsenate + thioredoxin the enzyme encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 reduces intracellular arsenate to the more toxic arsenite, which is subsequently extruded from the cell Staphylococcus aureus arsenite + thioredoxin disulfide + H2O
-
 ?
1.20.4.4 arsenate + thioredoxin the enzyme uses an intramolecular thiol pair (Cys82, Cys89) for the reduction of arsenate Staphylococcus aureus arsenite + thioredoxin disulfide + H2O
-
 ?
1.20.4.4 additional information assays are performed with different arsenate concentrations and arsenate reductase concentrations in the presence of 0.42 microM Escherichia coli thioredoxin, 0.14 microM Escherichia coli thioredoxin reductase and 125 microM NADPH Staphylococcus aureus ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.20.4.4 ? x * 14400, SDS-PAGE Staphylococcus aureus

Synonyms

EC Number Synonyms Comment Organism
1.20.4.4 ArsC
-
 Staphylococcus aureus
1.20.4.4 arsenate reductase
-
 Staphylococcus aureus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.20.4.4 37
-
 assay at Staphylococcus aureus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.20.4.4 0.061
-
 arsenate C15A ArsC mutant, at 2 microM AsO43- Staphylococcus aureus
1.20.4.4 0.061
-
 arsenate pH 7.5, 37°C, mutant enzyme C15A, at 0.002 mM arsenate Staphylococcus aureus
1.20.4.4 0.075
-
 arsenate wild-type ArsC, at 2 microM AsO43- Staphylococcus aureus
1.20.4.4 0.075
-
 arsenate pH 7.5, 37°C, wild-type enzyme, at 0.002 mM arsenate Staphylococcus aureus
1.20.4.4 0.08
-
 arsenate C15A ArsC mutant, at 10 mM AsO43- Staphylococcus aureus
1.20.4.4 0.08
-
 arsenate pH 7.5, 37°C, mutant enzyme C15A, at 10 mM arsenate Staphylococcus aureus
1.20.4.4 0.165
-
 arsenate wild-type ArsC, at 10 mM AsO43- Staphylococcus aureus
1.20.4.4 0.165
-
 arsenate pH 7.5, 37°C, wild-type enzyme, at 10 mM arsenate Staphylococcus aureus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.20.4.4 7.5
-
 assay at Staphylococcus aureus

Cofactor

EC Number Cofactor Comment Organism Structure
1.20.4.4 additional information ArsC couples to thioredoxin, thioredoxin reductase, and NADPH to be enzymatically active Staphylococcus aureus

General Information

EC Number General Information Comment Organism
1.20.4.4 physiological function the enzyme encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 reduces intracellular arsenate to the more toxic arsenite, which is subsequently extruded from the cell Staphylococcus aureus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.20.4.4 441
-
 arsenate pH 7.5, 37°C, mutant enzyme C15A, at 0.002 mM arsenate Staphylococcus aureus
1.20.4.4 455
-
 arsenate C15A mutant, 2.6 times lower Staphylococcus aureus
1.20.4.4 1136
-
 arsenate wild-type Staphylococcus aureus
1.20.4.4 1170
-
 arsenate pH 7.5, 37°C, wild-type enzyme, at 0.002 mM arsenate Staphylococcus aureus