Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
D407F | site-directed mutagenesis, mutant shows 8% of wild-type activity | Rattus norvegicus |
K399F | site-directed mutagenesis, mutant shows 28% of wild-type activity | Rattus norvegicus |
K399F/R400F/D407F | site-directed mutagenesis, triple mutant shows 10% of wild-type activity | Rattus norvegicus |
K399P/R400P/D407P | site-directed mutagenesis, triple mutant shows 10% of wild-type activity | Rattus norvegicus |
K399W/R400W/D407W | site-directed mutagenesis, inactive mutant | Rattus norvegicus |
additional information | construction of a truncated enzyme mutant | Rattus norvegicus |
R400F | site-directed mutagenesis, mutant shows 24% of wild-type activity | Rattus norvegicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
26-hydroxysqualene | competitive | Rattus norvegicus | |
NB-598 | squalenoid noncompetitive allylamine inhibitor | Rattus norvegicus | |
trisnorsqualene alcohol | squalenoid noncompetitive inhibitor, mechanism-based inactivator | Rattus norvegicus | |
trisnorsqualene alcohol diazoester | i.e. TNSA-Dza, competitive, able to photocovalently modify the native protein | Rattus norvegicus | |
trisnorsqualene cyclopropylamine | squalenoid noncompetitive inhibitor, mechanism-based inactivator | Rattus norvegicus | |
trisnorsqualene hydroxylamine | mechanism-based inactivator | Rattus norvegicus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
50000 | - |
2 * 50000, recombinant truncated mutant enzyme, SDS-PAGE | Rattus norvegicus |
100000 | - |
recombinant truncated mutant enzyme | Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Rattus norvegicus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
squalene + [reduced NADPH-hemoprotein reductase] + O2 = (3S)-2,3-epoxy-2,3-dihydrosqualene + [oxidized NADPH-hemoprotein reductase] + H2O | active site involves the key residues K399, R400, and D407 | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | location of the substrate-binding site by binding studies using the photolabeling inhibitor trisnorsqualene alcohol diazoester, key residues are K399, R400, and D407 | Rattus norvegicus | ? | - |
? | |
squalene + reduced acceptor + O2 | - |
Rattus norvegicus | (S)-squalene-2,3-epoxide + acceptor + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 50000, recombinant truncated mutant enzyme, SDS-PAGE | Rattus norvegicus |
Synonyms | Comment | Organism |
---|---|---|
SE | - |
Rattus norvegicus |
squalene epoxidase | - |
Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Rattus norvegicus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition kinetics | Rattus norvegicus | |
0.0184 | - |
trisnorsqualene alcohol diazoester | recombinant truncated enzyme mutant, pH 7.4, 37°C | Rattus norvegicus |