Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) pLysS | Oryctolagus cuniculus |
Protein Variants | Comment | Organism |
---|---|---|
F303Q | site-directed mutagenesis, the enzyme shows reduced 3-ketoreductase activity compared to the wild-type enzyme | Oryctolagus cuniculus |
F303Q/M304S | site-directed mutagenesis, the enzyme shows reduced 3-ketoreductase activity compared to the wild-type enzyme. The double mutation impairs the affinity and catalytic efficiency, although it did not affect the stereospecific reduction of the two 3-ketosteroids into the corresponding 3beta-hydroxysteroids | Oryctolagus cuniculus |
L116F | site-directed mutagenesis | Oryctolagus cuniculus |
M304S | site-directed mutagenesis, the enzyme shows reduced 3-ketoreductase activity compared to the wild-type enzyme. The M304S mutation causes a 4fold increase in the Km value for pyridine-3-aldehyde | Oryctolagus cuniculus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(Z)-2-(4-methoxyphenylimino)-7-hydroxy-N-(pyridin-2-yl)-2H-chromene-3-carboxamide | noncompetitive in the reduction reaction, competitive in the oxidation reaction | Oryctolagus cuniculus | |
3-(4-hydroxy-2-methoxyphenyl)acrylic acid 3-(3-hydroxyphenyl)propyl ester | uncompetitive in the reduction reaction, competitive in the oxidation reaction | Oryctolagus cuniculus | |
AL1567 | - |
Oryctolagus cuniculus | |
bisdemethoxycurcumin | - |
Oryctolagus cuniculus | |
diethylstilbestrol | - |
Oryctolagus cuniculus | |
Diphenic acid | competitive in the oxidation reaction | Oryctolagus cuniculus | |
epalrestat | - |
Oryctolagus cuniculus | |
minalrestat | competitive in the oxidation reaction | Oryctolagus cuniculus | |
oleanolic acid | competitive in the oxidation reaction | Oryctolagus cuniculus | |
quercetin | - |
Oryctolagus cuniculus | |
sorbinil | - |
Oryctolagus cuniculus | |
sulindac | - |
Oryctolagus cuniculus | |
Tolrestat | competitive in the oxidation reaction | Oryctolagus cuniculus | |
zopolrestat | - |
Oryctolagus cuniculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
NADPH | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.0023 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, mutant L116F | Oryctolagus cuniculus | |
0.0024 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.0035 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.0072 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, mutant L116F | Oryctolagus cuniculus | |
0.011 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, wild-type enzyme and mutant F303Q | Oryctolagus cuniculus | |
0.013 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, mutant F303Q | Oryctolagus cuniculus | |
0.017 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, mutant F303Q | Oryctolagus cuniculus | |
0.02 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, mutant M304S | Oryctolagus cuniculus | |
0.022 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, mutant L116F | Oryctolagus cuniculus | |
0.023 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, mutant M304S | Oryctolagus cuniculus | |
0.039 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, mutant F303Q/M304S | Oryctolagus cuniculus | |
0.039 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, mutant M304S | Oryctolagus cuniculus | |
0.048 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, mutant F303Q/M304S | Oryctolagus cuniculus | |
0.05 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, mutant F303Q/M304S | Oryctolagus cuniculus | |
0.18 | - |
methylglyoxal | pH 7.4, 25°C, mutant L116F | Oryctolagus cuniculus | |
0.209 | - |
methylglyoxal | pH 7.4, 25°C, mutant M304S | Oryctolagus cuniculus | |
0.222 | - |
methylglyoxal | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.25 | - |
methylglyoxal | pH 7.4, 25°C, mutant F303Q/M304S | Oryctolagus cuniculus | |
0.253 | - |
methylglyoxal | pH 7.4, 25°C, mutant F303Q | Oryctolagus cuniculus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Oryctolagus cuniculus | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
35000 | - |
native enzyme, gel filtration | Oryctolagus cuniculus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Oryctolagus cuniculus | the enzyme, termed AKR1B19, acts not only as a reductase for reactive carbonyl compounds derived from lipid peroxidation like AR-like proteins of other species, but also as a superior reductive 3beta-HSD for 3-keto-5alpha/beta-dihydro-C19/C21/C24-steroids | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Oryctolagus cuniculus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
native enzyme from lungs by ammonium sufate fractionation, gel filtration, and anion exchange chromatography, recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography and ultrafiltration | Oryctolagus cuniculus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
adrenal gland | - |
Oryctolagus cuniculus | - |
colon | - |
Oryctolagus cuniculus | - |
lung | - |
Oryctolagus cuniculus | - |
additional information | the enzyme expression occurs in all rabbit tissues, although its expression levels in the renal cortex, adrenal gland, small intestine and colon are high | Oryctolagus cuniculus | - |
renal cortex | - |
Oryctolagus cuniculus | - |
small intestine | - |
Oryctolagus cuniculus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5alpha-androstan-3,17-dione + NADPH + H+ | - |
Oryctolagus cuniculus | 5alpha-androstan-3beta-ol-17-one + NADP+ | - |
r | |
5alpha-dihydrotestosterone + NADPH + H+ | - |
Oryctolagus cuniculus | 3beta,17beta-dihydroxy-5alpha-androstane + NADP+ | - |
r | |
5alpha-pregnan-3,20-dione + NADPH + H+ | - |
Oryctolagus cuniculus | 5alpha-pregnane-3beta-ol-20-one + NADP+ | - |
r | |
5alpha-pregnane-21-ol-3,20-dione + NADPH + H+ | - |
Oryctolagus cuniculus | 5alpha-pregnane-3beta,21-diol-20-one + NADP+ | - |
r | |
5beta-androstan-3,17-dione + NADPH + H+ | - |
Oryctolagus cuniculus | 5beta-androstan-3beta-ol-17-one + NADP+ | - |
r | |
5beta-androstan-3beta-ol-17-one + NADP+ | - |
Oryctolagus cuniculus | 5beta-androstan-3,17-dione + NADPH + H+ | - |
r | |
5beta-androstane-3beta,17beta-diol + NADP+ | - |
Oryctolagus cuniculus | 5beta-androstane-17beta-ol-3-one + NADPH + H+ | - |
r | |
5beta-cholanic acid-3,7-dione + NADPH + H+ | - |
Oryctolagus cuniculus | 5beta-cholanic acid-3beta-ol-7-one + NADP+ | - |
r | |
5beta-dihydrocorticosterone + NADPH + H+ | - |
Oryctolagus cuniculus | (3beta,5beta,11beta)-3,11,21-trihydroxypregnan-20-one + NADP+ | - |
r | |
5beta-dihydrocortisone + NADPH + H+ | - |
Oryctolagus cuniculus | ? | - |
r | |
5beta-dihydrotestosterone + NADPH + H+ | - |
Oryctolagus cuniculus | 3beta,17beta-dihydroxy-5beta-androstane + NADP+ | - |
r | |
5beta-hydroxy-5beta cholanic acid + NADP+ | - |
Oryctolagus cuniculus | 3-oxo-5-beta-cholanic acid + NADPH + H+ | - |
r | |
5beta-pregnan-3,20-dione + NADPH + H+ | - |
Oryctolagus cuniculus | 5beta-pregnane-3beta-ol-20-one + NADP+ | - |
r | |
5beta-pregnane-20-ol-3-one + NADPH + H+ | - |
Oryctolagus cuniculus | 5beta-pregnane-3beta,20-diol + NADP+ | - |
r | |
5beta-pregnane-21-ol-3,20-dione + NADPH + H+ | - |
Oryctolagus cuniculus | 5beta-pregnane-3beta,21-diol-20-one + NADP+ | - |
r | |
5beta-pregnane-3beta,20alpha-diol + NADP+ | - |
Oryctolagus cuniculus | 5beta-pregnane-20alpha-ol-3-one + NADPH + H+ | - |
r | |
5beta-pregnane-3beta,20beta-diol + NADP+ | - |
Oryctolagus cuniculus | 5beta-pregnane-20beta-ol-3-one + NADPH + H+ | - |
r | |
5beta-pregnane-3beta,21-diol-20-one + NADP+ | - |
Oryctolagus cuniculus | 5beta-pregnane-21-ol-3,20-dione + NADPH + H+ | - |
r | |
5beta-pregnane-3beta-ol-20-one + NADP+ | - |
Oryctolagus cuniculus | 5beta-pregnan-3,20-dione + NADPH + H+ | - |
r | |
dehydrolithocholic acid + NADPH + H+ | - |
Oryctolagus cuniculus | ? | - |
r | |
farnesol + NADP+ | - |
Oryctolagus cuniculus | farnesal + NADPH + H+ | - |
? | |
geranylgeraniol + NADP+ | - |
Oryctolagus cuniculus | geranylgeranial + NADPH + H+ | - |
? | |
isolithocholic acid + NADP+ | - |
Oryctolagus cuniculus | ? + NADPH + H+ | - |
r | |
methylglyoxal + NADP+ | - |
Oryctolagus cuniculus | ? + NADPH + H+ | - |
? | |
additional information | the enzyme, termed AKR1B19, acts not only as a reductase for reactive carbonyl compounds derived from lipid peroxidation like AR-like proteins of other species, but also as a superior reductive 3beta-HSD for 3-keto-5alpha/beta-dihydro-C19/C21/C24-steroids | Oryctolagus cuniculus | ? | - |
? | |
additional information | substrate specificity of the bifunctional enzyme, for carbonyl compounds and steroids, overview. No reductase activity is observed for 17- and 20-ketosteroids, DELTA4-3-ketosteroids (testosterone, 4-androstene-3,17-dione and progesterone), and prostaglandins (D2, E2, and A1). In the reverse reaction, the enzyme oxidizes 3beta-hydroxy-5alpha/beta-dihydrosteroids, but shows no significant dehydrogenase activity for DELTA5-3beta-hydroxysteroids (dehydroepiandrosterone, pregnenolone, and 5-pregnene-3beta,20alpha-diol) and 3alpha-hydroxysteroids (5alpha/beta-androstan-3alpha-ol-17-ones, 5alpha/beta-androstane-3alpha,17beta-diols, 5alpha/beta-pregnan-3alpha-ol-20-ones and lithocholic acid). The reactivity towards all-trans-retinal of AKR1B19 is low | Oryctolagus cuniculus | ? | - |
? | |
pyridine-3-aldehyde + NADP+ | - |
Oryctolagus cuniculus | pyridin-3-ylmethanol + NADPH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
reductive 3beta-HSD | - |
Oryctolagus cuniculus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Oryctolagus cuniculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, mutant F303Q/M304S | Oryctolagus cuniculus | |
0.023 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, mutant F303Q/M304S | Oryctolagus cuniculus | |
0.042 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, mutant M304S | Oryctolagus cuniculus | |
0.047 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, mutant M304S | Oryctolagus cuniculus | |
0.062 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, mutant F303Q | Oryctolagus cuniculus | |
0.063 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, mutant L116F | Oryctolagus cuniculus | |
0.08 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, mutant F303Q | Oryctolagus cuniculus | |
0.092 | - |
5beta-pregnane-21-ol-3,20-dione | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.107 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, mutant L116F | Oryctolagus cuniculus | |
0.113 | - |
5beta-dihydrotestosterone | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.3 | - |
methylglyoxal | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.32 | - |
methylglyoxal | pH 7.4, 25°C, mutant L116F | Oryctolagus cuniculus | |
0.37 | - |
methylglyoxal | pH 7.4, 25°C, mutant M304S | Oryctolagus cuniculus | |
0.4 | - |
methylglyoxal | pH 7.4, 25°C, mutant F303Q/M304S | Oryctolagus cuniculus | |
0.58 | - |
methylglyoxal | pH 7.4, 25°C, mutant F303Q | Oryctolagus cuniculus | |
0.6 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, mutant L116F | Oryctolagus cuniculus | |
0.72 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.73 | - |
NADPH | pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | |
0.78 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, mutant M304S | Oryctolagus cuniculus | |
0.87 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, mutant F303Q | Oryctolagus cuniculus | |
1.2 | - |
Pyridine-3-aldehyde | pH 7.4, 25°C, mutant F303Q/M304S | Oryctolagus cuniculus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.2 | 6.4 | - |
Oryctolagus cuniculus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | the NADH-linked reductase activity is only 18% of the NADPH-linked activity | Oryctolagus cuniculus | |
NADPH | AKR1B19 shows high coenzyme preference to NADPH | Oryctolagus cuniculus |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.000008 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | (Z)-2-(4-methoxyphenylimino)-7-hydroxy-N-(pyridin-2-yl)-2H-chromene-3-carboxamide | |
0.000043 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | minalrestat | |
0.00006 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | 3-(4-hydroxy-2-methoxyphenyl)acrylic acid 3-(3-hydroxyphenyl)propyl ester | |
0.00016 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | Tolrestat | |
0.0004 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | oleanolic acid | |
0.0006 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | bisdemethoxycurcumin | |
0.00086 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | sulindac | |
0.0012 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | zopolrestat | |
0.0015 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | quercetin | |
0.0015 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | epalrestat | |
0.0017 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | AL1567 | |
0.0028 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | diethylstilbestrol | |
0.0053 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | sorbinil | |
0.26 | - |
pH 7.4, 25°C, wild-type enzyme | Oryctolagus cuniculus | Diphenic acid |
General Information | Comment | Organism |
---|---|---|
evolution | the rabbit aldose reductase-like protein that shars an 86% sequence identity to human aldo-keto reductase (AKR) 1B10 and is assigned as AKR1B19 in the AKR superfamily. It is bifunctional and also acts as a 3-ketoreductase reducing 3-keto-5alpha/beta-dihydro-C19/C21/C24-steroids into the corresponding 3betahydroxysteroids | Oryctolagus cuniculus |
malfunction | single and double mutations, F303Q, M304S and F303Q/M304S, significantly impair the 3-ketoreductase activity, suggesting that the two residues play critical roles in recognition of the steroidal substrate | Oryctolagus cuniculus |
physiological function | the rabbit aldose reductase-like protein, assigned as AKR1B19, is bifunctional, it shows aldose reductase activity for various aldehydes and alpha-dicarbonyl compounds acting as a defense system against cytotoxic carbonyl compounds in rabbit tissues, and it also acts as a 3-ketoreductase reducing 3-keto-5alpha/beta-dihydro-C19/C21/C24-steroids into the corresponding 3beta-hydroxysteroids | Oryctolagus cuniculus |