Cloned (Comment) | Organism |
---|---|
gene gsto1, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis and tree, recombinant overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Nilaparvata lugens |
Protein Variants | Comment | Organism |
---|---|---|
C29A | site-directed mutagenesis, the mutant shows 8.1fold lower activity with dehydroascorbate compared to wild-type | Nilaparvata lugens |
F28A | site-directed mutagenesis, the mutant shows 2.6fold lower activity with dehydroascorbate compared to wild-type | Nilaparvata lugens |
F30L | site-directed mutagenesis, the mutant shows 6.8fold lower activity with dehydroascorbate compared to wild-type | Nilaparvata lugens |
L225A | site-directed mutagenesis, the mutant shows 3.8fold lower activity with dehydroascorbate compared to wild-type | Nilaparvata lugens |
R176A | site-directed mutagenesis, the mutant shows 3.6fold lower activity with dehydroascorbate compared to wild-type | Nilaparvata lugens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
diazinon | about 40% inhibition | Nilaparvata lugens | |
permethrin | about 40% inhibition | Nilaparvata lugens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | the DHA reaction follows Michaelis-Menten kinetics | Nilaparvata lugens | |
0.32 | - |
dehydroascorbate | recombinant mutant L225A, pH 8.0, 30°C | Nilaparvata lugens | |
0.55 | - |
dehydroascorbate | recombinant mutant F28A, pH 8.0, 30°C | Nilaparvata lugens | |
0.6 | - |
dehydroascorbate | recombinant wild-type enzyme, pH 8.0, 30°C | Nilaparvata lugens | |
0.6 | - |
dehydroascorbate | recombinant mutant R176A, pH 8.0, 30°C | Nilaparvata lugens | |
0.67 | - |
dehydroascorbate | recombinant mutant P30A, pH 8.0, 30°C | Nilaparvata lugens | |
0.77 | - |
dehydroascorbate | recombinant mutant C29A, pH 8.0, 30°C | Nilaparvata lugens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 glutathione + dehydroascorbate | Nilaparvata lugens | - |
glutathione disulfide + ascorbate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Nilaparvata lugens | J9Q3Y4 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Nilaparvata lugens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.1 | - |
purified recombinant wild-type enzyme, pH 8.0, 30°C | Nilaparvata lugens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 glutathione + dehydroascorbate | - |
Nilaparvata lugens | glutathione disulfide + ascorbate | - |
? | |
additional information | enzyme nlGSTO shows broad substrate specificity as thiol transferase, see also EC 2.5.1.18 | Nilaparvata lugens | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
dehydroascorbate reductase | - |
Nilaparvata lugens |
DHA reductase | - |
Nilaparvata lugens |
GSTO1 | - |
Nilaparvata lugens |
More | omega-class glutathione S-transferase | Nilaparvata lugens |
nlGSTO | - |
Nilaparvata lugens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Nilaparvata lugens |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
purified recombinant enzyme, stable up to | Nilaparvata lugens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.029 | - |
dehydroascorbate | recombinant mutant L225A, pH 8.0, 30°C | Nilaparvata lugens | |
0.033 | - |
dehydroascorbate | recombinant mutant C29A, pH 8.0, 30°C | Nilaparvata lugens | |
0.035 | - |
dehydroascorbate | recombinant mutant P30A, pH 8.0, 30°C | Nilaparvata lugens | |
0.04 | - |
dehydroascorbate | recombinant mutant R176A, pH 8.0, 30°C | Nilaparvata lugens | |
0.074 | - |
dehydroascorbate | recombinant mutant F28A, pH 8.0, 30°C | Nilaparvata lugens | |
0.21 | - |
dehydroascorbate | recombinant wild-type enzyme, pH 8.0, 30°C | Nilaparvata lugens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Nilaparvata lugens |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | 8 | purified recombinant nlGSTO retains more than 75% of its original activity | Nilaparvata lugens |
General Information | Comment | Organism |
---|---|---|
metabolism | nlGSTO exhibits DHA reductase and thiol transferase, which are responsible for antioxidant reactions. GSH is known to be a redox regulator. Thiol transferase is involved in GSH homeostasis, which is important for antioxidant defense. DHA reductase is responsible for maintaining the balance of ascorbate, which functions in scavenging reactive oxygen species | Nilaparvata lugens |
additional information | putative substrate-binding sites, including Phe28, Cys29, Phe30, Arg176, and Lue225, are important for glutathione transferase and dehydroascorbate reductase activities | Nilaparvata lugens |
physiological function | the omega-class glutathione S-transferase (GST), nlGSTO, of the brown planthopper, Nilaparvata lugens, catalyzes the biotransformation of glutathione with 1-chloro-2,4-dinitrobenzene, a general substrate for GST, as well as with dehydroascorbate to synthesize ascorbate. As ascorbate is a reducing agent, nlGSTO may participate in antioxidant resistance | Nilaparvata lugens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.043 | - |
dehydroascorbate | recombinant mutant C29A, pH 8.0, 30°C | Nilaparvata lugens | |
0.052 | - |
dehydroascorbate | recombinant mutant P30A, pH 8.0, 30°C | Nilaparvata lugens | |
0.067 | - |
dehydroascorbate | recombinant mutant R176A, pH 8.0, 30°C | Nilaparvata lugens | |
0.092 | - |
dehydroascorbate | recombinant mutant L225A, pH 8.0, 30°C | Nilaparvata lugens | |
0.135 | - |
dehydroascorbate | recombinant mutant F28A, pH 8.0, 30°C | Nilaparvata lugens | |
0.35 | - |
dehydroascorbate | recombinant wild-type enzyme, pH 8.0, 30°C | Nilaparvata lugens |