Cloned (Comment) | Organism |
---|---|
wild-type and mutant bmGST expression in Escherichia coli strain Rosetta (DE3)pLysS | Bombyx mori |
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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Bombyx mori | - |
- |
- |
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 | 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 | 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 | Comment | Organism |
---|---|---|
bmGSTD | - |
Bombyx mori |
Delta-class glutathione transferase | - |
Bombyx mori |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Bombyx mori |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Bombyx mori |
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 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 |