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Literature summary for 2.5.1.18 extracted from
- Structural analysis of an epsilon-class glutathione transferase from housefly, Musca domestica (2013), Biochem. Biophys. Res. Commun., 430, 1206-1211.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of His6-tagged isozyme MdGST6B in Escherichia coli strain BL21 CodonPlus (DE3)-RIL | Musca domestica |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant detagged isozyme MdGST6B complexed with reduced glutathione, hanging drop vapor diffusion method, 10 mg/ml protein in 10 mM Tris-HCl, pH 8.0, and 10 mM GSH is mixed in a 1:1 ratio with a reservoir solution containing 0.1 M HEPES-NaOH, pH 7.0, and 1.2 M Na citrate, 20°C, several days, X-ray diffraction structure determination and analysis at 1.8 A resolution | Musca domestica |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| F108G | site-directed mutagenesis, the mutant shows reduced affinity and catalytic activity to both GSH and the electrophilic co-substrate, 1-chloro-2,4-dinitrobenzene, compared to the wild-type enzyme | Musca domestica |
| R113A | site-directed mutagensis, the mutant shows activity slightly reduced affinity and catalytic activity to both GSH and the electrophilic co-substrate, 1-chloro-2,4-dinitrobenzene, compared to the wild-type enzyme | Musca domestica |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0067 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged wild-type enzyme, pH and temperature not specified in the publication | Musca domestica |  |
| 0.0084 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged mutant R113A, pH and temperature not specified in the publication | Musca domestica | |
| 0.0088 | - |
1-chloro-2,4-dinitrobenzene | recombinant detagged wild-type enzyme, pH and temperature not specified in the publication | Musca domestica | |
| 0.0728 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged mutant F108G, pH and temperature not specified in the publication | Musca domestica | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Musca domestica | Q9U795 | two epsilon-class GSTs MdGST6A and MdGST6B | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N-terminally His6-tagged isozyme MdGST6B from Escherichia coli strain BL21 CodonPlus (DE3)-RIL by nickel affinity chromatography | Musca domestica |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1-chloro-2,4-dinitrobenzene + glutathione | - |
Musca domestica | S-(2,4-dinitrophenyl)glutathione + HCl | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | isozyme MdGST6B shows typical GST folding comprised of N-terminal and C-terminal domains | Musca domestica |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| epsilon-class glutathione transferase | - |
Musca domestica |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.99 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged mutant F108G, pH and temperature not specified in the publication | Musca domestica | |
| 1.13 | - |
glutathione | mutant F108G, pH and temperature not specified in the publication | Musca domestica | |
| 8 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged mutant R113A, pH and temperature not specified in the publication | Musca domestica | |
| 11 | - |
glutathione | mutant R113A, pH and temperature not specified in the publication | Musca domestica | |
| 11.7 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged wild-type enzyme, pH and temperature not specified in the publication | Musca domestica | |
| 14 | - |
glutathione | wild-type enzyme, pH and temperature not specified in the publication | Musca domestica | |
| 25.1 | - |
1-chloro-2,4-dinitrobenzene | recombinant detagged wild-type enzyme, pH and temperature not specified in the publication | Musca domestica | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| malfunction | mutation at Phe108, located just below Arg113 in the binding pocket, reduces the affinity and catalytic activity to both GSH and the electrophilic co-substrate, 1-chloro-2,4-dinitrobenzene | Musca domestica |
| additional information | the substrate binding pocket of MdGST6B, involving Arg113 and Phe121 on helix 4, is narrower compared to other delta- and epsilon-class GSTs, structure, overview. The Arg113 hydrogen bond does not play a crucial role in catalysis | Musca domestica |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.014 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged mutant F108G, pH and temperature not specified in the publication | Musca domestica | |
| 0.95 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged mutant R113A, pH and temperature not specified in the publication | Musca domestica | |
| 1.73 | - |
1-chloro-2,4-dinitrobenzene | recombinant His-tagged wild-type enzyme, pH and temperature not specified in the publication | Musca domestica | |
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