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

BRENDA support

Literature summary for 2.5.1.18 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
Search for more literature for 2.5.1.18

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

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

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Bombyx mori
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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)

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

Subunits

Subunits Comment Organism
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

Synonyms Comment Organism
bmGSTD
-
 Bombyx mori
Delta-class glutathione transferase
-
 Bombyx mori

Temperature Optimum [°C]

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

pH Optimum

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

General Information

General Information Comment Organism
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]

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