Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type enzyme complexed with inhibitors phenylacetic acid, 2-hydroxyphenylacetic acid, 2,6-dichlorophenylacetic acid, hexanoic acid, and lipoic acid, hanging drop vapor diffusion method, 4°C, 23-25 mg/ml protein in 0.10 M sodium phosphate buffer, pH 7.0, mixed with well solution containing 20% w/v PEG 6000 and 50 mM sodium citrate, pH 5.0, microseeding, 1 month, crystal stabilization by 0.1 to 1.0% glutaraldehyde, X-ray diffraction structure determination and analysis at 1.7-2.0 A resolution | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C298A/W219Y | site-directed mutagenesis, altered kinetics and thermodynamics in phenylacetic acid derivative inhibitor binding compared to the wild-type enzyme | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2,6-dichlorophenylacetic acid | competitive | Homo sapiens | |
2-bromophenylacetic acid | competitive | Homo sapiens | |
2-Chlorophenylacetic acid | competitive | Homo sapiens | |
2-hydroxyphenylacetic acid | competitive | Homo sapiens | |
2-naphthylacetic acid | competitive | Homo sapiens | |
DL-dihydrolipoic acid | binds very tight to the enzyme, best inhibitor of the investigated, competitive | Homo sapiens | |
DL-lipoamide | binds three-fold more weakly than D,L-lipoic acid, competitive | Homo sapiens | |
DL-lipoic acid | is effective in the treatment of diabetic complications, binds tightly to the enzyme, competitive | Homo sapiens | |
hexanoic acid | competitive | Homo sapiens | |
Mandelic acid | competitive | Homo sapiens | |
additional information | enzyme-inhibitor complex structures, the hydrophobic nature of the enzyme active site makes significant contributions to inhibitor binding rather than a reliance on pi-pi orbital interactions with Trp 20, overview | Homo sapiens | |
phenylacetic acid | competitive | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
35727 | - |
x * 35727, sequence calculation | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glyceraldehyde + NADPH + H+ | - |
Homo sapiens | glycerol + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 35727, sequence calculation | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
AKR1B1 | - |
Homo sapiens |
aldose reductase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition kinetics and thermodynamic studies of inhibitors binding to the enzyme, overview | Homo sapiens |