Cloned (Comment) | Organism |
---|---|
expression of His-tagged enzyme in Escherichia coli | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant hGSTk in apo-form and in complex with S-hexylglutathione, hanging drop vapour diffusion method, apo-form of hGSTk in 20 mM NaH2PO4, pH 7.4, 20 mM NaCl, 1 mM EDTA and 7.2 mM 2-mercaptoethanol, hGSTk in complex in 20 mM HEPES, pH 7.0, 50 mM NaCl, 1 mM EDTA and 1 mM DTT, supplemented with S-hexylglutathione at a molar ratio of 1:2, is mixed with an equal volume of reservoir solution containing 0.2 M NaSCN and 20% PEG3350, 20°C, X-ray diffraction structure determination and analysis at 1.8-1.9 A resolution | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D201A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
D69A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
D69T | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
F83A | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
F83W | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
F87A | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
F87W | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
I44A | site-directed mutagenesis, H-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
K62A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
K62E | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
L15A | site-directed mutagenesis, H-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
L183A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
L58A | site-directed mutagenesis, H-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
L88A | site-directed mutagenesis, H-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
L92A | site-directed mutagenesis, H-site mutation, inactive mutant | Homo sapiens |
M66A | site-directed mutagenesis, H-site mutation, inactive mutant | Homo sapiens |
M91A | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
M91K | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
N53A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
P17A | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
P55A | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
P56A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
P56A | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
R202A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
S16A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
S19A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
S200A | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
W126A | site-directed mutagenesis, H-site mutation, the mutant shows increased activity compared to the wild-type enzyme | Homo sapiens |
Y18A | site-directed mutagenesis, G-site mutation, inactive mutant | Homo sapiens |
Y18F | site-directed mutagenesis, G-site mutation, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
Y18H | site-directed mutagenesis, G-site mutation, inactive mutant | Homo sapiens |
Y18L | site-directed mutagenesis, G-site mutation, inactive mutant | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | recombinant hGSTk, steady-state kinetics, overview | Homo sapiens | |
0.17 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant W126A | Homo sapiens | |
0.19 | - |
glutathione | pH 7.4, 25°C, recombinant wild-type GSTk | Homo sapiens | |
0.34 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P17A | Homo sapiens | |
0.62 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant M91A | Homo sapiens | |
0.63 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant M91K | Homo sapiens | |
0.64 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F83W | Homo sapiens | |
0.74 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P56A | Homo sapiens | |
0.91 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F87A | Homo sapiens | |
1.03 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S200A | Homo sapiens | |
1.08 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P55A | Homo sapiens | |
1.1 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant wild-type GSTk | Homo sapiens | |
1.25 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant K62A | Homo sapiens | |
1.3 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F83A | Homo sapiens | |
1.38 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F87W | Homo sapiens | |
1.45 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L58A | Homo sapiens | |
1.61 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S19A | Homo sapiens | |
1.68 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant P56A | Homo sapiens | |
2.01 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant I44A | Homo sapiens | |
2.59 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L15A | Homo sapiens | |
4.44 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L88A | Homo sapiens | |
4.53 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant N53A | Homo sapiens | |
4.74 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D201A | Homo sapiens | |
9.81 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant L183A | Homo sapiens | |
17.34 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S16A | Homo sapiens | |
20.45 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant Y18F | Homo sapiens | |
25.26 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant K62E | Homo sapiens | |
60.89 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D69T | Homo sapiens | |
105.4 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D69A | Homo sapiens | |
123.6 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant R202A | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrial matrix | - |
Homo sapiens | 5759 | - |
peroxisome | matrix | Homo sapiens | 5777 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and dialysis | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
RX + glutathione = HX + R-S-glutathione | catalytic mechanism of kappa class GST, overview. Substrate binding induces a conformational change of the active site from an open conformation in the apo-form to a closed conformation in the S-hexylglutathione-bound complex, facilitating formations of the G site (GSH-binding site) and the H site (hydrophobic substrate-binding site). The conserved Ser16 at the G site functions as the catalytic residue in the deprotonation of the thiol group and the conserved Asp69, Ser200,Asp201 andArg202 form a network of interactions with gamma -glutamyl carboxylate to stabilize the thiolate anion. The H site is a large hydrophobic pocket with conformational flexibility to allow the binding of different hydrophobic substrates. The kinetic mechanism of hGSTk conforms to a rapid equilibrium random sequential Bi Bi model | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-chloro-2,4-dinitrobenzene + glutathione | - |
Homo sapiens | 2,4-dinitrophenyl-glutathione + HCl | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | crystal structure | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
GSTk | - |
Homo sapiens |
kappa class glutathione transferase | - |
Homo sapiens |
Kappa class GST | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S16A | Homo sapiens | |
0.32 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D69A | Homo sapiens | |
0.47 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D201A | Homo sapiens | |
0.47 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant L183A | Homo sapiens | |
0.67 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant R202A | Homo sapiens | |
0.75 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant K62E | Homo sapiens | |
0.89 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant K62A | Homo sapiens | |
0.99 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S200A | Homo sapiens | |
1.18 | - |
glutathione | pH 7.4, 25°C, recombinant wild-type GSTk | Homo sapiens | |
1.19 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L15A | Homo sapiens | |
1.33 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D69T | Homo sapiens | |
1.56 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant Y18F | Homo sapiens | |
1.75 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant N53A | Homo sapiens | |
2.29 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant P56A | Homo sapiens | |
2.38 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P17A | Homo sapiens | |
2.4 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L58A | Homo sapiens | |
2.94 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant W126A | Homo sapiens | |
3.18 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F83W | Homo sapiens | |
3.28 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant I44A | Homo sapiens | |
3.74 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S19A | Homo sapiens | |
3.94 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant wild-type GSTk | Homo sapiens | |
4.18 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F87A | Homo sapiens | |
4.5 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P55A | Homo sapiens | |
5 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L88A | Homo sapiens | |
5.11 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant M91K | Homo sapiens | |
5.5 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant M91A | Homo sapiens | |
5.78 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F87W | Homo sapiens | |
6.24 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F83A | Homo sapiens | |
7.83 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P56A | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
evolution | kappa class GSTs are a distinct class of the GST superfamily or Trx (thioredoxin)-fold superfamily because of their unique cellular localization, function and structure. GSTks do not contain a conserved CXXC motif at the active site | Homo sapiens |
additional information | G-site and H-site structure analysis and modeling, overview | Homo sapiens |
physiological function | glutathione transferases are a family of enzymes that primarily catalyse nucleophilic addition of the thiol of GSH to a variety of hydrophobic electrophiles in the cellular detoxification of cytotoxic and genotoxic compounds | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00304 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D69A | Homo sapiens | |
0.00542 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant R202A | Homo sapiens | |
0.0098 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S16A | Homo sapiens | |
0.0218 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D69T | Homo sapiens | |
0.0297 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant K62E | Homo sapiens | |
0.0479 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant L183A | Homo sapiens | |
0.0763 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant Y18F | Homo sapiens | |
0.0992 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant D201A | Homo sapiens | |
0.232 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S19A | Homo sapiens | |
0.386 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant N53A | Homo sapiens | |
0.712 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant K62A | Homo sapiens | |
0.737 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L15A | Homo sapiens | |
0.961 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant S200A | Homo sapiens | |
1.13 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L88A | Homo sapiens | |
1.36 | - |
glutathione | pH 7.4, 25°C, recombinant GSTk mutant P56A | Homo sapiens | |
1.63 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant I44A | Homo sapiens | |
1.66 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant L58A | Homo sapiens | |
3.58 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant wild-type GSTk | Homo sapiens | |
4.17 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P55A | Homo sapiens | |
4.19 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F87W | Homo sapiens | |
4.59 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F87A | Homo sapiens | |
4.8 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F83A | Homo sapiens | |
4.97 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant F83W | Homo sapiens | |
6.21 | - |
glutathione | pH 7.4, 25°C, recombinant wild-type GSTk | Homo sapiens | |
7 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P17A | Homo sapiens | |
8.11 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant M91K | Homo sapiens | |
8.87 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant M91A | Homo sapiens | |
10.6 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant P56A | Homo sapiens | |
17.3 | - |
1-chloro-2,4-dinitrobenzene | pH 7.4, 25°C, recombinant GSTk mutant W126A | Homo sapiens |