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Literature summary for 2.5.1.18 extracted from

  • Lerksuthirat, T.; Ketterman, A.J.
    Characterization of putative hydrophobic substrate binding site residues of a Delta class glutathione transferase from Anopheles dirus (2008), Arch. Biochem. Biophys., 479, 97-103.
    View publication on PubMed
Search for more literature for 2.5.1.18

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)pLysS cells Anopheles dirus

Protein Variants

Protein Variants Comment Organism
F212A decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
F212E decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
F212H decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
F212L decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
F212W decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
F212Y decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
F216A decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
Q112A slightly decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
R214A decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
T174A decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
Y215A decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
Y215E decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
Y215F increased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
Y215H decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus
Y215S decreased activity towards 1-chloro-2,4-dinitrobenzene compared to the wild type enzyme Anopheles dirus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.01
-
 Ethacrynic acid mutant enzyme Y215S, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.032
-
 Ethacrynic acid mutant enzyme Y215A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.037
-
 Ethacrynic acid mutant enzyme Q112A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.065
-
 Ethacrynic acid wild type enzyme, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.1
-
 Ethacrynic acid mutant enzyme F212L, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.33
-
 glutathione mutant enzyme T174A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.35
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215F, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.38
-
 glutathione mutant enzyme F212E, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.44
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212L, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.45
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Q112A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.48
-
 glutathione mutant enzyme F212Y, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.52
-
 1-chloro-2,4-dinitrobenzene mutant enzyme R214A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.53
-
 glutathione mutant enzyme F212H, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.53
-
 1-chloro-2,4-dinitrobenzene wild type enzyme, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.6
-
 glutathione mutant enzyme Q112A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.62
-
 glutathione mutant enzyme R214A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.64
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215H, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.65
-
 glutathione mutant enzyme F212A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.67
-
 glutathione mutant enzyme F212W, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.7
-
 glutathione wild type enzyme, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.73
-
 glutathione mutant enzyme Y215A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.76
-
 glutathione mutant enzyme Y215E, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.78
-
 glutathione mutant enzyme Y215S, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.85
-
 glutathione mutant enzyme F216A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.85
-
 glutathione mutant enzyme Y215F, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.88
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215E, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.93
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.99
-
 glutathione mutant enzyme Y215H, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
1.01
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215S, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
1.77
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212Y, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
1.78
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212W, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
1.92
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F216A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
2.03
-
 glutathione mutant enzyme F212L, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
2.35
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212H, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
2.37
-
 1-chloro-2,4-dinitrobenzene mutant enzyme T174A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
3.06
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
3.69
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212E, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus

Organism

Organism UniProt Comment Textmining
Anopheles dirus Q9GN60
-
-

Purification (Commentary)

Purification (Comment) Organism
GSTrap affinity column chromatography Anopheles dirus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.005
-
 mutant enzyme F212H, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.005
-
 mutant enzyme Y215E, using 4-nitrophenethyl bromide as a substrate Anopheles dirus
0.006
-
 mutant enzyme T174A, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.006
-
 mutant enzyme Y215A, using 4-nitrophenethyl bromide as a substrate Anopheles dirus
0.009
-
 mutant enzyme F212Y, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.01
-
 mutant enzyme F212H, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.012
-
 mutant enzyme F212Y, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.017
-
 mutant enzyme F212W, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.017
-
 mutant enzyme R214A, using 4-nitrophenethyl bromide as a substrate Anopheles dirus
0.018
-
 mutant enzyme Q112A, using 4-nitrophenethyl bromide as a substrate Anopheles dirus
0.019
-
 mutant enzyme Y215H, using 4-nitrophenethyl bromide as a substrate Anopheles dirus
0.021
-
 mutant enzyme F212W, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.025
-
 mutant enzyme Y215A, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.026
-
 mutant enzyme Y215S, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.029
-
 mutant enzyme Y215E, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.029
-
 mutant enzyme Y215S, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.03
-
 mutant enzyme Y215A, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.03
-
 wild type enzyme, using 4-nitrophenethyl bromide as a substrate Anopheles dirus
0.034
-
 mutant enzyme F212A, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.035
-
 mutant enzyme Y215E, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.036
-
 mutant enzyme Q112A, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.036
-
 mutant enzyme Y215A, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.04
-
 mutant enzyme Y215H, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.041
-
 mutant enzyme F212A, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.043
-
 mutant enzyme Y215F, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.044
-
 mutant enzyme Q112A, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.048
-
 mutant enzyme R214A, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.049
-
 mutant enzyme Y215A, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.052
-
 mutant enzyme R214A, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.053
-
 mutant enzyme Y215H, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.059
-
 mutant enzyme F212L, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.068
-
 wild type enzyme, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.069
-
 mutant enzyme Y215F, using 4-nitrophenethyl bromide as a substrate Anopheles dirus
0.073
-
 mutant enzyme T174A, using ethacrynic acid as a substrate Anopheles dirus
0.079
-
 wild type enzyme, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.085
-
 mutant enzyme F212A, using ethacrynic acid as a substrate Anopheles dirus
0.086
-
 mutant enzyme F212E, using ethacrynic acid as a substrate Anopheles dirus
0.086
-
 mutant enzyme Y215F, using 4-nitrobenzyl chloride as a substrate Anopheles dirus
0.087
-
 mutant enzyme F212L, using 1,2-dichloro-4-nitrobenzene as a substrate Anopheles dirus
0.12
-
 mutant enzyme F212H, using ethacrynic acid as a substrate Anopheles dirus
0.17
-
 mutant enzyme Y215H, using ethacrynic acid as a substrate Anopheles dirus
0.29
-
 mutant enzyme Y215F, using ethacrynic acid as a substrate Anopheles dirus
0.3
-
 mutant enzyme Y215E, using ethacrynic acid as a substrate Anopheles dirus
0.31
-
 mutant enzyme F212Y, using ethacrynic acid as a substrate Anopheles dirus
0.34
-
 mutant enzyme F212L, using ethacrynic acid as a substrate Anopheles dirus
0.37
-
 mutant enzyme F212W, using ethacrynic acid as a substrate Anopheles dirus
0.47
-
 mutant enzyme R214A, using ethacrynic acid as a substrate Anopheles dirus
0.47
-
 mutant enzyme Y215A, using ethacrynic acid as a substrate Anopheles dirus
0.51
-
 mutant enzyme F212E, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
0.57
-
 wild type enzyme, using ethacrynic acid as a substrate Anopheles dirus
0.61
-
 mutant enzyme Q112A, using ethacrynic acid as a substrate Anopheles dirus
0.64
-
 mutant enzyme Y215S, using ethacrynic acid as a substrate Anopheles dirus
1.03
-
 mutant enzyme Y215A, using ethacrynic acid as a substrate Anopheles dirus
3.03
-
 mutant enzyme T174A, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
4.28
-
 mutant enzyme F212H, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
8.03
-
 mutant enzyme F212Y, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
12.2
-
 mutant enzyme F212A, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
12.3
-
 mutant enzyme F212W, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
22.9
-
 mutant enzyme Y215A, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
25.2
-
 mutant enzyme Y215A, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
26.1
-
 mutant enzyme Y215E, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
27
-
 mutant enzyme Y215S, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
32.2
-
 mutant enzyme F212L, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
36.2
-
 mutant enzyme Y215H, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
44.3
-
 mutant enzyme R214A, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
52.4
-
 mutant enzyme Q112A, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
58.6
-
 wild type enzyme, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus
74.3
-
 mutant enzyme Y215F, using 1-chloro-2,4-dinitrobenzene as a substrate Anopheles dirus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glutathione + 1,2-dichloro-4-nitrobenzene weak substrate Anopheles dirus ?
-
 ?
glutathione + 1-chloro-2,4-dinitrobenzene best substrate Anopheles dirus S-(2,4-dinitrophenyl)-glutathione + HCl
-
 ?
glutathione + 4-nitrobenzyl chloride weak substrate Anopheles dirus ?
-
 ?
glutathione + 4-nitrophenethyl bromide weak substrate Anopheles dirus ?
-
 ?
glutathione + ethacrynic acid weak substrate Anopheles dirus ?
-
 ?

Synonyms

Synonyms Comment Organism
GSTD4-4
-
 Anopheles dirus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 the wild type enzyme shows a half-life of 13.3 min at 45°C Anopheles dirus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.2
-
 Ethacrynic acid mutant enzyme F212L, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.28
-
 Ethacrynic acid wild type enzyme, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.36
-
 Ethacrynic acid mutant enzyme Q112A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.38
-
 Ethacrynic acid mutant enzyme Y215S, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.4
-
 Ethacrynic acid mutant enzyme Y215A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
0.54
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212E, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
2.14
-
 1-chloro-2,4-dinitrobenzene mutant enzyme T174A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
3.55
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212H, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
6.44
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212Y, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
8.85
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212W, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
12.6
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215S, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
12.8
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F216A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
13.9
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
14.9
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
17
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215E, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
18.7
-
 1-chloro-2,4-dinitrobenzene mutant enzyme F212L, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
20.5
-
 1-chloro-2,4-dinitrobenzene mutant enzyme R214A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
20.7
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215H, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
26.4
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Q112A, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
26.5
-
 1-chloro-2,4-dinitrobenzene wild type enzyme, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus
35.7
-
 1-chloro-2,4-dinitrobenzene mutant enzyme Y215F, in 0.1 M phosphate buffer pH 6.5 Anopheles dirus