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Literature summary for 1.1.1.270 extracted from

  • Endo, S.; Matsunaga, T.; Kumada, S.; Fujimoto, A.; Ohno, S.; El-Kabbani, O.; Hu, D.; Toyooka, N.; Mano, J.; Tajima, K.; Hara, A.
    Characterization of rabbit aldose reductase-like protein with 3beta-hydroxysteroid dehydrogenase activity (2012), Arch. Biochem. Biophys., 527, 23-30.
    View publication on PubMed

Cloned(Commentary)

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

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

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 [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

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Oryctolagus cuniculus 5829
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
native enzyme, gel filtration Oryctolagus cuniculus

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Oryctolagus cuniculus
-
-
-

Purification (Commentary)

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

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 and Products (Substrate)

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

Synonyms Comment Organism
reductive 3beta-HSD
-
Oryctolagus cuniculus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Oryctolagus cuniculus

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.2 6.4
-
Oryctolagus cuniculus

Cofactor

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 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

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