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Literature summary for 1.1.1.213 extracted from
- Characterization of the substrate binding site in rat liver 3alpha-hydroxysteroid/dihydrodiol dehydrogenase. The roles of tryptophans in ligand binding and protein fluorescence (1996), J. Biol. Chem., 271, 30190-30198.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| wild-type enzyme and mutants are overexpressed in Escherichia coli DH5alpha | Rattus norvegicus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| W148Y | site-directed mutagenesis, kinetically similar to wild type enzyme, no function in ligand binding | Rattus norvegicus |
| W227Y | site-directed mutagenesis, important role in binding steroid hormones, but not small substrates or inhibitors, interacts with the C and/or D-rings of steroid ligand | Rattus norvegicus |
| W86Y | site-directed mutagenesis, important in binding steroids and non-steroidal anti-inflamatory drugs, region in which it resides is part of the substrate/inhibitor binding pocket, near the A-, and B-rings of bound steroid | Rattus norvegicus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| (E/Z)-sulfindac | wild-type, W86Y and W227Y | Rattus norvegicus |  |
| 1,10-phenanthroline | wild-type, W86Y and W227Y | Rattus norvegicus | |
| 1,7-phenanthroline | wild-type, W86Y and W227Y | Rattus norvegicus | |
| Flufenamic acid | wild-type, W86Y and W227Y | Rattus norvegicus | |
| Hexestrol | - |
Rattus norvegicus | |
| indomethacin | wild-type, W86Y and W227Y | Rattus norvegicus | |
| Meclofenamic acid | wild-type, W86Y and W227Y | Rattus norvegicus | |
| Mefenamic acid | wild-type, W86Y and W227Y | Rattus norvegicus | |
| non-steroidal anti-inflamatory drug | - |
Rattus norvegicus | |
| ponalrestat | wild-type, W86Y and W227Y, weak inhibitor | Rattus norvegicus | |
| zopolrestat | wild-type, W86Y and W227Y, weak inhibitor | Rattus norvegicus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00122 | 0.0051 | 9,10-phenanthrenequinone | - |
Rattus norvegicus | |
| 0.042 | 0.408 | 3alpha-hydroxy-5alpha-androstan-17-one | - |
Rattus norvegicus | |
| 0.163 | 0.211 | 4-nitrobenzaldehyde | - |
Rattus norvegicus | |
| 3.54 | 33.7 | 2-decalone | - |
Rattus norvegicus | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 17beta-hydroxy-5alpha-androstan-3-one + NAD(P)H + H+ | Rattus norvegicus | - |
3alpha,17beta-dihydroxy-5alpha-androstan + NAD(P)+ | - |
? | |
| additional information | Rattus norvegicus | inactivating circulating androgens, progestins and glucocorticoids. Regulation of hormone levels in endocrine target tissues and in carcinogen actibation | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Rattus norvegicus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| anion exchange and Blue Sepharose column | Rattus norvegicus |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ | aldoketoreductase superfamily | Rattus norvegicus | |
| a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ | bi bi mechanism, pyridine nucleotide binds before substrate | Rattus norvegicus |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| liver | - |
Rattus norvegicus | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.059 | - |
W86Y mutant | Rattus norvegicus |
| 0.08 | - |
W227Y mutant | Rattus norvegicus |
| 0.674 | - |
W148Y mutant | Rattus norvegicus |
| 1.5 | - |
native enzyme | Rattus norvegicus |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -70°C, 20 mM potassium phosphate buffer, pH 7.0, 1 mM EDTA, 1 mM beta-mercaptoethanol, 30% glycerol | Rattus norvegicus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 17beta-hydroxy-5alpha-androstan-3-one + NAD(P)H + H+ | - |
Rattus norvegicus | 3alpha,17beta-dihydroxy-5alpha-androstan + NAD(P)+ | - |
? | |
| 2-decalone + NADH | pH 6.0 | Rattus norvegicus | ? + NAD+ | - |
? | |
| 3alpha-hydroxy-5alpha-androstan-17-one + NAD+ | - |
Rattus norvegicus | 5alpha-androstan-3,17-dione + NADH | - |
r | |
| 4-nitrobenzaldehyde + NAD(P)H | pH 6.0 | Rattus norvegicus | (4-nitrophenyl)methanol + NAD(P)+ | - |
? | |
| 4-nitrobenzaldehyde + NAD(P)H | NADH as cofactor | Rattus norvegicus | (4-nitrophenyl)methanol + NAD(P)+ | - |
? | |
| 9,10-phenanthrenequinone + NAD(P)H | NADPH | Rattus norvegicus | ? + NAD(P)+ | - |
? | |
| 9,10-phenanthrenequinone + NAD(P)H | pH 6.0 | Rattus norvegicus | ? + NAD(P)+ | - |
? | |
| additional information | inactivating circulating androgens, progestins and glucocorticoids. Regulation of hormone levels in endocrine target tissues and in carcinogen actibation | Rattus norvegicus | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | (alpha/beta)8-barrel fold | Rattus norvegicus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.17 | 3.93 | 9,10-phenanthrenequinone | - |
Rattus norvegicus | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | 4-pro-R-hydride transfer of the nicotinamide cofactor | Rattus norvegicus | |
| NAD+ | - |
Rattus norvegicus | |
| NADH | - |
Rattus norvegicus | |
| NADPH | - |
Rattus norvegicus | |
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