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Literature summary for 2.5.1.18 extracted from

  • Yamamoto, K.; Higashiura, A.; Hossain, M.T.; Yamada, N.; Shiotsuki, T.; Nakagawa, A.
    Structural characterization of the catalytic site of a Nilaparvata lugens delta-class glutathione transferase (2015), Arch. Biochem. Biophys., 566, 36-42.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta (DE3) pLysS cells Nilaparvata lugens

Crystallization (Commentary)

Crystallization (Comment) Organism
in complex with glutathione, sitting drop vapor diffusion method, using 0.1 M Tris-HCl (pH 8.0) containing 1 M LiCl and 30% (w/v) PEG-6000 Nilaparvata lugens

Protein Variants

Protein Variants Comment Organism
E66A the mutant shows reduced activity compared to the wild type enzyme Nilaparvata lugens
F119A the mutant shows reduced activity compared to the wild type enzyme Nilaparvata lugens
H52A the mutant shows reduced activity compared to the wild type enzyme Nilaparvata lugens
R68A the mutant shows reduced activity compared to the wild type enzyme Nilaparvata lugens
S11A the mutant shows reduced activity compared to the wild type enzyme Nilaparvata lugens
S67A the mutant shows reduced activity compared to the wild type enzyme Nilaparvata lugens
V54A the mutant shows reduced activity compared to the wild type enzyme Nilaparvata lugens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
glutathione wild type enzyme, at pH 8.0 and 30°C Nilaparvata lugens
0.19
-
glutathione mutant enzyme S67A, at pH 8.0 and 30°C Nilaparvata lugens
0.26
-
1-chloro-2,4-dinitrobenzene mutant enzyme S11A, at pH 8.0 and 30°C Nilaparvata lugens
0.41
-
glutathione mutant enzyme F119A, at pH 8.0 and 30°C Nilaparvata lugens
0.42
-
1-chloro-2,4-dinitrobenzene mutant enzyme F119A, at pH 8.0 and 30°C Nilaparvata lugens
0.46
-
1-chloro-2,4-dinitrobenzene mutant enzyme V54A, at pH 8.0 and 30°C Nilaparvata lugens
0.73
-
glutathione mutant enzyme H52A, at pH 8.0 and 30°C Nilaparvata lugens
0.74
-
glutathione mutant enzyme S11A, at pH 8.0 and 30°C Nilaparvata lugens
0.84
-
1-chloro-2,4-dinitrobenzene wild type enzyme, at pH 8.0 and 30°C Nilaparvata lugens
0.93
-
1-chloro-2,4-dinitrobenzene mutant enzyme R68A, at pH 8.0 and 30°C Nilaparvata lugens
1
-
1-chloro-2,4-dinitrobenzene mutant enzyme H52A, at pH 8.0 and 30°C Nilaparvata lugens
1
-
1-chloro-2,4-dinitrobenzene mutant enzyme S67A, at pH 8.0 and 30°C Nilaparvata lugens
1.2
-
glutathione mutant enzyme V54A, at pH 8.0 and 30°C Nilaparvata lugens
1.7
-
1-chloro-2,4-dinitrobenzene mutant enzyme E66A, at pH 8.0 and 30°C Nilaparvata lugens
3.3
-
glutathione mutant enzyme R68A, at pH 8.0 and 30°C Nilaparvata lugens
3.4
-
glutathione mutant enzyme E66A, at pH 8.0 and 30°C Nilaparvata lugens

Organism

Organism UniProt Comment Textmining
Nilaparvata lugens Q8N0B3 strain Izumo
-

Purification (Commentary)

Purification (Comment) Organism
DEAE-Sepharose column chromatography and Superdex 200 gel filtration Nilaparvata lugens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-chloro-2,4-dinitrobenzene + glutathione
-
Nilaparvata lugens S-(2,4-dinitrophenyl)glutathione + HCl
-
?
additional information no activity with 4-hydroxynonenal, ethacrynic acid, 4-nitrobenzyl chloride, 1,2-epoxy-3-(4-nitrophenoxy)-propane, 4-nitrophenethyl bromide, and 4-nitrophenyl acetate Nilaparvata lugens ?
-
?

Subunits

Subunits Comment Organism
homodimer x-ray crystallography Nilaparvata lugens

Synonyms

Synonyms Comment Organism
Delta-class glutathione transferase
-
Nilaparvata lugens
GSTD
-
Nilaparvata lugens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
the enzyme is stable at temperatures below 50°C Nilaparvata lugens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.16
-
glutathione mutant enzyme S11A, at pH 8.0 and 30°C Nilaparvata lugens
0.27
-
1-chloro-2,4-dinitrobenzene mutant enzyme S11A, at pH 8.0 and 30°C Nilaparvata lugens
0.33
-
1-chloro-2,4-dinitrobenzene mutant enzyme F119A, at pH 8.0 and 30°C Nilaparvata lugens
0.52
-
glutathione mutant enzyme F119A, at pH 8.0 and 30°C Nilaparvata lugens
0.58
-
glutathione mutant enzyme H52A, at pH 8.0 and 30°C Nilaparvata lugens
0.78
-
glutathione mutant enzyme V54A, at pH 8.0 and 30°C Nilaparvata lugens
0.85
-
glutathione mutant enzyme E66A, at pH 8.0 and 30°C Nilaparvata lugens
1.07
-
1-chloro-2,4-dinitrobenzene mutant enzyme V54A, at pH 8.0 and 30°C Nilaparvata lugens
1.27
-
1-chloro-2,4-dinitrobenzene mutant enzyme H52A, at pH 8.0 and 30°C Nilaparvata lugens
1.62
-
glutathione wild type enzyme, at pH 8.0 and 30°C Nilaparvata lugens
1.83
-
glutathione mutant enzyme S67A, at pH 8.0 and 30°C Nilaparvata lugens
2.67
-
1-chloro-2,4-dinitrobenzene wild type enzyme, at pH 8.0 and 30°C Nilaparvata lugens
2.67
-
1-chloro-2,4-dinitrobenzene mutant enzyme E66A, at pH 8.0 and 30°C Nilaparvata lugens
2.67
-
glutathione mutant enzyme R68A, at pH 8.0 and 30°C Nilaparvata lugens
3
-
1-chloro-2,4-dinitrobenzene mutant enzyme S67A, at pH 8.0 and 30°C Nilaparvata lugens
3.17
-
1-chloro-2,4-dinitrobenzene mutant enzyme R68A, at pH 8.0 and 30°C Nilaparvata lugens

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 12 the enzyme retains more than 80% of its original activity when shifted from pH 6.0 to 12.0 Nilaparvata lugens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0000217
-
glutathione mutant enzyme S11A, at pH 8.0 and 30°C Nilaparvata lugens
0.000025
-
glutathione mutant enzyme E66A, at pH 8.0 and 30°C Nilaparvata lugens
0.000065
-
glutathione mutant enzyme V54A, at pH 8.0 and 30°C Nilaparvata lugens
0.00008
-
1-chloro-2,4-dinitrobenzene mutant enzyme F119A, at pH 8.0 and 30°C Nilaparvata lugens
0.00008
-
glutathione mutant enzyme H52A, at pH 8.0 and 30°C Nilaparvata lugens
0.0000817
-
glutathione mutant enzyme R68A, at pH 8.0 and 30°C Nilaparvata lugens
0.000103
-
1-chloro-2,4-dinitrobenzene mutant enzyme S11A, at pH 8.0 and 30°C Nilaparvata lugens
0.000127
-
glutathione mutant enzyme F119A, at pH 8.0 and 30°C Nilaparvata lugens
0.000127
-
1-chloro-2,4-dinitrobenzene mutant enzyme H52A, at pH 8.0 and 30°C Nilaparvata lugens
0.000153
-
1-chloro-2,4-dinitrobenzene mutant enzyme E66A, at pH 8.0 and 30°C Nilaparvata lugens
0.0023
-
1-chloro-2,4-dinitrobenzene mutant enzyme V54A, at pH 8.0 and 30°C Nilaparvata lugens
0.003
-
1-chloro-2,4-dinitrobenzene mutant enzyme S67A, at pH 8.0 and 30°C Nilaparvata lugens
0.0032
-
1-chloro-2,4-dinitrobenzene wild type enzyme, at pH 8.0 and 30°C Nilaparvata lugens
0.0033
-
1-chloro-2,4-dinitrobenzene mutant enzyme R68A, at pH 8.0 and 30°C Nilaparvata lugens
0.009
-
glutathione wild type enzyme, at pH 8.0 and 30°C Nilaparvata lugens
0.0098
-
glutathione mutant enzyme S67A, at pH 8.0 and 30°C Nilaparvata lugens