Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Yamamoto, K.; Usuda, K.; Kakuta, Y.; Kimura, M.; Higashiura, A.; Nakagawa, A.; Aso, Y.; Suzuki, M.
    Structural basis for catalytic activity of a silkworm Delta-class glutathione transferase (2012), Biochim. Biophys. Acta, 1820, 1469-1474.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.18 wild-type and mutant bmGST expression in Escherichia coli strain Rosetta (DE3)pLysS Bombyx mori

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.18 purified bmGSTD, sitting-drop vapor diffusion method, 10 mg/ml protein in 20 mM Tris-HCl buffer, pH 8.0, and 0.2 M NaCl, is mixed with with 0.2 M sodium acetate trihydrate, 0.1 M sodium cacodylate trihydrate, pH 6.5, and 30% w/v PEG 8000, 20°C, 1 month, three-dimensional structure of bmGSTD by the molecular replacement method to a resolution of 2.0 A resolution Bombyx mori

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.18 H52A site-directed mutagensis, the mutant shows reduced activity compared to the wild-type enzyme Bombyx mori
2.5.1.18 Q51A site-directed mutagensis, the mutant shows reduced activity compared to the wild-type enzyme Bombyx mori
2.5.1.18 R68A site-directed mutagensis, inactive mutant Bombyx mori
2.5.1.18 S11A site-directed mutagensis, the mutant shows reduced activity compared to the wild-type enzyme Bombyx mori
2.5.1.18 S67A site-directed mutagensis, the mutant shows reduced activity compared to the wild-type enzyme Bombyx mori

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.18 0.21
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant mutant S11A Bombyx mori
2.5.1.18 0.48
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant wild-type enzyme Bombyx mori
2.5.1.18 0.52
-
 glutathione pH 6.5, 30°C, recombinant wild-type enzyme Bombyx mori
2.5.1.18 0.6
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant mutant Q51A Bombyx mori
2.5.1.18 0.68
-
 glutathione pH 6.5, 30°C, recombinant mutant H52A Bombyx mori
2.5.1.18 1.19
-
 glutathione pH 6.5, 30°C, recombinant mutant S11A Bombyx mori
2.5.1.18 1.22
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant mutant H52A Bombyx mori
2.5.1.18 1.39
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant mutant S67A Bombyx mori
2.5.1.18 2.33
-
 glutathione pH 6.5, 30°C, recombinant mutant Q51A Bombyx mori
2.5.1.18 2.7
-
 glutathione pH 6.5, 30°C, recombinant mutant S67A Bombyx mori

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.18 Bombyx mori
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.18 recombinant wild-type and mutant bmGST from Escherichia coli strain Rosetta (DE3)pLysS by anion exchange chromatography and gel filtration Bombyx mori

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.18 1-chloro-2,4-dinitrobenzene + glutathione synthetic substrate of GST Bombyx mori S-(2,4-dinitrophenyl)glutathione + HCl
-
 ?

Subunits

EC Number Subunits Comment Organism
2.5.1.18 More bmGSTD contains two distinct domains, an N-terminal domain and a C-terminal domain, connected by a linker. The N-terminal domain binds glutathione Bombyx mori

Synonyms

EC Number Synonyms Comment Organism
2.5.1.18 bmGSTD
-
 Bombyx mori
2.5.1.18 Delta-class glutathione transferase
-
 Bombyx mori

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.18 30
-
 assay at Bombyx mori

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.18 6.5
-
 assay at Bombyx mori

General Information

EC Number General Information Comment Organism
2.5.1.18 additional information residues Ser11, Gln51, His52, Ser67, and Arg68 of the N-terminal domain are important for catalytic enzyme function, active site structure, overview Bombyx mori

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.5.1.18 0.0017
-
 glutathione pH 6.5, 30°C, recombinant mutant S11A Bombyx mori
2.5.1.18 0.007
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant mutant S11A Bombyx mori
2.5.1.18 0.013
-
 glutathione pH 6.5, 30°C, recombinant mutant S67A Bombyx mori
2.5.1.18 0.028
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant mutant S67A Bombyx mori
2.5.1.18 0.095
-
 glutathione pH 6.5, 30°C, recombinant mutant Q51A Bombyx mori
2.5.1.18 0.237
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant mutant Q51A Bombyx mori
2.5.1.18 0.253
-
 glutathione pH 6.5, 30°C, recombinant mutant H52A Bombyx mori
2.5.1.18 0.293
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant mutant H52A Bombyx mori
2.5.1.18 3.3
-
 glutathione pH 6.5, 30°C, recombinant wild-type enzyme Bombyx mori
2.5.1.18 4.47
-
 1-chloro-2,4-dinitrobenzene pH 6.5, 30°C, recombinant wild-type enzyme Bombyx mori