Cloned (Comment) | Organism |
---|---|
gene Atu3701, DNA and amino acid sequence determination and analysis, sequence comparison and phylogenetic analysis, expression of His-tagged wild-type and mutant AtuGSTH1-1s in Escherichia coli strain BL21 (DE3) | Agrobacterium tumefaciens |
Crystallization (Comment) | Organism |
---|---|
AtuGSTH1-1 in complex with inhibitor S-(4-nitrobenzyl)-glutathione, hanging drop vapor diffusion method, mixing of 0.002 ml of 4.85 mg/ml protein in 15 mM Tris-HCl, pH 7.0, and 10 mM S-(4-nitrobenzyl)-glutathione, with 0.002 ml of well solution 1.4 M Na/K phosphate, pH 8.3, equilibration against 0.8 ml well solution, 16°C, X-ray diffraction structure determination and analysis at 1.4 A resolution | Agrobacterium tumefaciens |
Protein Variants | Comment | Organism |
---|---|---|
F22A | site-directed mutagenesis, the mutant shows reduced glutathione transferase activity and increased glutatione peroxidase activity compared to the wild-type GSTH1-1 | Agrobacterium tumefaciens |
R187A | site-directed mutagenesis, the mutant shows reduced glutathione transferase activity and reduced glutatione peroxidase activity compared to the wild-type GSTH1-1 | Agrobacterium tumefaciens |
R34A | site-directed mutagenesis, almost inactive mutant | Agrobacterium tumefaciens |
S25A | site-directed mutagenesis, the mutant shows reduced glutathione transferase activity and increased glutatione peroxidase activity compared to the wild-type GSTH1-1 | Agrobacterium tumefaciens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
S-(p-nitrobenzyl)-glutathione | - |
Agrobacterium tumefaciens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state Michaelis-Menten kinetic analysis of AtuGSTH1-1 | Agrobacterium tumefaciens | |
0.085 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1 mutant F22A, with 1-chloro-2,4-dinitrobenzene | Agrobacterium tumefaciens | |
0.14 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1 mutant S25A, with 1-chloro-2,4-dinitrobenzene | Agrobacterium tumefaciens | |
0.3 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1, with 1-chloro-2,4-dinitrobenzene | Agrobacterium tumefaciens | |
0.34 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1 mutant R187A, with 1-chloro-2,4-dinitrobenzene | Agrobacterium tumefaciens | |
1.5 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 37°C, recombinant AtuGSTH1-1 | Agrobacterium tumefaciens | |
1.7 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1, with 2-hydroxyethyl disulfide | Agrobacterium tumefaciens | |
1.9 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 37°C, recombinant AtuGSTH1-1 mutant S25A | Agrobacterium tumefaciens | |
3.2 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 37°C, recombinant AtuGSTH1-1 mutant F22A | Agrobacterium tumefaciens | |
4.1 | - |
2-hydroxyethyl disulfide | pH 6.5, 37°C, recombinant AtuGSTH1-1 | Agrobacterium tumefaciens | |
7.1 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 37°C, recombinant AtuGSTH1-1 mutant R187A | Agrobacterium tumefaciens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Agrobacterium tumefaciens | 5829 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Agrobacterium tumefaciens | - |
gene Atu3701 encoding an eta class GST, i.e. AtuGSTH1-1 | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | - |
gene Atu3701 encoding an eta class GST, i.e. AtuGSTH1-1 | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant AtuGSTH1-1s from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography | Agrobacterium tumefaciens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
RX + glutathione = HX + R-S-glutathione | catalytic mechanism of isozyme GSTH1-1 | Agrobacterium tumefaciens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-chloro-2,4-dinitrobenzene + glutathione | - |
Agrobacterium tumefaciens | S-(2,4-dinitrophenyl)glutathione + HCl | - |
? | |
1-chloro-2,4-dinitrobenzene + glutathione | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | S-(2,4-dinitrophenyl)glutathione + HCl | - |
? | |
2-hydroxyethyl disulfide + glutathione | - |
Agrobacterium tumefaciens | ? | - |
? | |
2-hydroxyethyl disulfide + glutathione | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | ? | - |
? | |
additional information | AtuGSTH1-1 exhibits significant transferase activity against the common substrates aryl halides, as well as very high peroxidase activity towards organic hydroperoxides | Agrobacterium tumefaciens | ? | - |
? | |
additional information | AtuGSTH1-1 exhibits significant transferase activity against the common substrates aryl halides, as well as very high peroxidase activity towards organic hydroperoxides | Agrobacterium tumefaciens C58 / ATCC 33970 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AtuGSTH1-1 | - |
Agrobacterium tumefaciens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Agrobacterium tumefaciens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
2-hydroxyethyl disulfide | pH 6.5, 37°C, recombinant AtuGSTH1-1 | Agrobacterium tumefaciens | |
0.04 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1, with 2-hydroxyethyl disulfide | Agrobacterium tumefaciens | |
0.53 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 37°C, recombinant AtuGSTH1-1 | Agrobacterium tumefaciens | |
0.53 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1, with 1-chloro-2,4-dinitrobenzene | Agrobacterium tumefaciens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Agrobacterium tumefaciens |
General Information | Comment | Organism |
---|---|---|
additional information | AtuGSTH1-1 lacks the classic catalytic essential residues (Tyr, Ser, Cys) and has instead of the classic catalytic residues, an Arg residue (Arg34), an electron-sharing network, and a bridge of a network of water molecules may form the basis of the catalytic mechanism, Phe22, Ser25, and Arg187 are additional important residues for the enzyme's catalytic efficiency and specificity, overview. GSH binding site, G-site, and electrophilic binding site, H-site, structures, overview. In contrast to other GSTs where the H-site involves C-terminal domain residues, interactions in AtuGSTH1-1 are mainly provided by Arg187 and the long turn between strand b1 andhelix H1, residues 25-33 | Agrobacterium tumefaciens |
physiological function | AtuGSTH1-1 may play an important regulatory role in stress defence mechanism | Agrobacterium tumefaciens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
2-hydroxyethyl disulfide | pH 6.5, 37°C, recombinant AtuGSTH1-1 | Agrobacterium tumefaciens | |
0.01 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1, with 2-hydroxyethyl disulfide | Agrobacterium tumefaciens | |
0.36 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 37°C, recombinant AtuGSTH1-1 | Agrobacterium tumefaciens | |
0.36 | - |
glutathione | pH 6.5, 37°C, recombinant AtuGSTH1-1, with 1-chloro-2,4-dinitrobenzene | Agrobacterium tumefaciens |