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Literature summary extracted from

  • Vergauwen, B.; Pauwels, F.; Jacquemotte, F.; Meyer, T.E.; Cusanovich, M.A.; Bartsch, R.G.; van Beeumen, J.J.
    Characterization of glutathione amide reductase from Chromatium gracile. Identification of a novel thiol peroxidase (Prx/Grx) fueled by glutathione amide redox cycling (2001), J. Biol. Chem., 276, 20890-20897.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.8.1.16 expression in Escherichia coli Marichromatium gracile
1.11.1.17 expressed in Escherichia coli BL(DE3) Marichromatium gracile

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.8.1.16 0.0132
-
 NADH pH 7.1, 25°C Marichromatium gracile
1.8.1.16 0.097
-
 glutathione amide disulfide pH 7.1, 25°C Marichromatium gracile
1.8.1.16 1.98
-
 NADPH pH 7.1, 25°C Marichromatium gracile
1.8.1.16 6.9
-
 glutathione disulfide pH 7.1, 25°C Marichromatium gracile
1.11.1.17 additional information
-
 additional information kinetic analysis of the glutathione amide reductase, using glutathione amide disulfide (cf. EC 1.8.1.16) and NADH as substrates Marichromatium gracile

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.8.1.16 49028
-
 2 * 49028, calculation from sequence Marichromatium gracile
1.8.1.16 49030
-
 2 * 49030, nanospray ionization mass spectrometry Marichromatium gracile
1.8.1.16 98000
-
 gel filtration Marichromatium gracile
1.11.1.17 27430
-
 calculated from the deduced amino acid sequence Marichromatium gracile
1.11.1.17 27500
-
 chimeric enzyme composed of one N-terminal peroxidoxin-like domain followed by a glutaredoxin-like C-terminus, SDS-PAGE Marichromatium gracile

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.8.1.16 glutathione amide disulfide + NADH + H+ Marichromatium gracile
-
 2 glutathione amide + NAD+
-
 ?
1.8.1.16 glutathione amide disulfide + NADH + H+ Marichromatium gracile DSM 1712
-
 2 glutathione amide + NAD+
-
 ?
1.8.1.16 glutathione amide disulfide + NADPH + H+ Marichromatium gracile
-
 2 glutathione amide + NADP+
-
 ?
1.8.1.16 glutathione amide disulfide + NADPH + H+ Marichromatium gracile DSM 1712
-
 2 glutathione amide + NADP+
-
 ?
1.8.1.16 glutathione disulfide + NADH + H+ Marichromatium gracile
-
 2 glutathione + NAD+
-
 ?
1.8.1.16 glutathione disulfide + NADH + H+ Marichromatium gracile DSM 1712
-
 2 glutathione + NAD+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.8.1.16 Marichromatium gracile
-
-
-
1.11.1.17 Marichromatium gracile
-
-
-
1.11.1.17 Marichromatium gracile DSM 1712
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.8.1.16
-
 Marichromatium gracile
1.11.1.17 partial purification of the Prx/Grx-containing crude extract, centrifugation, ammonium sulfate precipitation, butyl-Sepharose HR 16/10 column, dialysis, ResourceQ column Marichromatium gracile

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.8.1.16 glutathione amide disulfide + NADH + H+
-
 Marichromatium gracile 2 glutathione amide + NAD+
-
 ?
1.8.1.16 glutathione amide disulfide + NADH + H+
-
 Marichromatium gracile DSM 1712 2 glutathione amide + NAD+
-
 ?
1.8.1.16 glutathione amide disulfide + NADPH + H+
-
 Marichromatium gracile 2 glutathione amide + NADP+
-
 ?
1.8.1.16 glutathione amide disulfide + NADPH + H+
-
 Marichromatium gracile DSM 1712 2 glutathione amide + NADP+
-
 ?
1.8.1.16 glutathione disulfide + NADH + H+
-
 Marichromatium gracile 2 glutathione + NAD+
-
 ?
1.8.1.16 glutathione disulfide + NADH + H+
-
 Marichromatium gracile DSM 1712 2 glutathione + NAD+
-
 ?
1.11.1.17 glutathione amide + H2O2 similar acitivity with small alkyl hydroperoxides indicating broad substrate specificity Marichromatium gracile glutathione amide disulfide + H2O
-
 ?
1.11.1.17 glutathione amide + H2O2 similar acitivity with small alkyl hydroperoxides indicating broad substrate specificity Marichromatium gracile DSM 1712 glutathione amide disulfide + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.8.1.16 dimer 2 * 49028, calculation from sequence Marichromatium gracile
1.8.1.16 dimer 2 * 49030, nanospray ionization mass spectrometry Marichromatium gracile

Synonyms

EC Number Synonyms Comment Organism
1.8.1.16 GAR
-
 Marichromatium gracile
1.8.1.16 garB
-
 Marichromatium gracile
1.11.1.17 garB
-
 Marichromatium gracile
1.11.1.17 More glutathione amide reductase, cf. EC 1.8.1.16 Marichromatium gracile
1.11.1.17 Prx-containing peroxidase
-
 Marichromatium gracile
1.11.1.17 Prx/Grx chimeric enzyme peroxidoxin/glutaredoxin chimeric enzyme Marichromatium gracile

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.11.1.17 25
-
 assay at Marichromatium gracile

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.8.1.16 0.14
-
 NADPH pH 7.1, 25°C Marichromatium gracile
1.8.1.16 146
-
 glutathione disulfide pH 7.1, 25°C Marichromatium gracile
1.8.1.16 186.8
-
 NADH pH 7.1, 25°C Marichromatium gracile
1.8.1.16 249.7
-
 glutathione amide disulfide pH 7.1, 25°C Marichromatium gracile
1.11.1.17 additional information
-
 additional information kinetic analysis of the glutathione amide reductase, using glutathione amide disulfide (cf. EC 1.8.1.16) and NADH as substrates Marichromatium gracile

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.11.1.17 7.1
-
 assay at Marichromatium gracile

Cofactor

EC Number Cofactor Comment Organism Structure
1.8.1.16 FAD formation of a charge-transfer complex between one of the newly reduced active site thiols and the oxidized FAD Marichromatium gracile
1.8.1.16 NADH preferred cofactor Marichromatium gracile
1.11.1.17 FAD
-
 Marichromatium gracile

General Information

EC Number General Information Comment Organism
1.11.1.17 physiological function garB encodes the central enzyme in glutathione amide cycling. The enzyme is the first example of a prokaryotic low molecular mass thiol-dependent peroxidase. The hydroperoxide-dependent Prx/Grx-catalyzed oxidation of glutathione amide is established in vitro, and as to be expected, the reaction is fueled by NADH via glutathione amide redox recycling Marichromatium gracile

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.8.1.16 0.07
-
 NADPH pH 7.1, 25°C Marichromatium gracile
1.8.1.16 21.2
-
 glutathione disulfide pH 7.1, 25°C Marichromatium gracile
1.8.1.16 2574
-
 glutathione amide disulfide pH 7.1, 25°C Marichromatium gracile
1.8.1.16 14150
-
 NADH pH 7.1, 25°C Marichromatium gracile
1.11.1.17 additional information
-
 additional information kinetic analysis of the glutathione amide reductase, using glutathione amide disulfide (cf. EC 1.8.1.16) and NADH as substrates Marichromatium gracile