Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain JM105 | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
D97K | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
D97R | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
E100K | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
E100R | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
additional information | construction of the monomeric mutant isozyme M1, GST M1, by introducing point mutations in the electrostatic region of the subunit interface and residues Arg77, Asp97, Glu100, Asn101, the mutation leads to formation of monomers instead of dimers and partially reduced activity, at 3 M KBr GST M1 has a specific activity close to that of GST M1-1, overview | Rattus norvegicus |
N101D | site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme | Rattus norvegicus |
N101K | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
R77E | site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme | Rattus norvegicus |
R77Q | site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme | Rattus norvegicus |
General Stability | Organism |
---|---|
72 h, 10°C, pH 6.5, 0-3 M KBr, purified recombinant wild-type enzyme | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.019 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, recombinant wild-type enzyme | Rattus norvegicus | |
0.02 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, recombinant mutant R77Q | Rattus norvegicus | |
0.045 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, recombinant mutant R77E | Rattus norvegicus | |
0.052 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, recombinant mutant D97R | Rattus norvegicus | |
0.069 | - |
glutathione | pH 6.5, 25°C, recombinant wild-type enzyme | Rattus norvegicus | |
0.077 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, recombinant mutant E100K | Rattus norvegicus | |
0.088 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, recombinant mutant D97K | Rattus norvegicus | |
0.089 | - |
glutathione | pH 6.5, 25°C, recombinant mutant R77Q | Rattus norvegicus | |
0.096 | - |
glutathione | pH 6.5, 25°C, recombinant mutant R77E | Rattus norvegicus | |
0.23 | - |
glutathione | pH 6.5, 25°C, recombinant mutant D97K | Rattus norvegicus | |
0.36 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, recombinant mutant N101D | Rattus norvegicus | |
0.36 | - |
1-chloro-2,4-dinitrobenzene | pH 6.5, 25°C, recombinant mutant N101K | Rattus norvegicus | |
0.39 | - |
glutathione | pH 6.5, 25°C, recombinant mutant N101D | Rattus norvegicus | |
0.56 | - |
glutathione | pH 6.5, 25°C, recombinant mutant D97R | Rattus norvegicus | |
0.78 | - |
glutathione | pH 6.5, 25°C, recombinant mutant E100R | Rattus norvegicus | |
2.3 | - |
glutathione | pH 6.5, 25°C, recombinant mutant E100K | Rattus norvegicus | |
6.6 | - |
glutathione | pH 6.5, 25°C, recombinant mutant N101K | Rattus norvegicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
KBr | KBr greatly influences the monomer-dimer equilibrium of the wildtype isozyme GST-M1-1 | Rattus norvegicus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
additional information | - |
monomer-dimer equilibrium and average molecular weights with and without bound glutathione at different concentrations of KBr, overview | Rattus norvegicus |
50000 | - |
about, recombinant wild-type enzyme, analytical ultracentrifugation | Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
mu class isozyme M1-1, GST-M1-1 | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glutathione + 1-chloro-2,4-dinitrobenzene | i.e. CDNB | Rattus norvegicus | chloride + 2,4-dinitrophenyl-glutathione | - |
? | |
additional information | the wild-type dimeric form is not required for catalytic activity | Rattus norvegicus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | wild-type isozyme GST-M1-1, homodimer, crystal structure determination | Rattus norvegicus |
monomer | mutant isozyme GST-M1, crystal structure determination | Rattus norvegicus |
More | molecular structure modeling, the wild-type dimeric form is not required for catalytic activity, overview | Rattus norvegicus |
Synonyms | Comment | Organism |
---|---|---|
GST | - |
Rattus norvegicus |
mu glutathione transferase | - |
Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Rattus norvegicus |