Protein Variants | Comment | Organism |
---|---|---|
R276M | site-directed mutagenesis, mutant does no longer form a salt-linkage to the phosphate of 2'-AMP, and does no longer bind tightly to NAD(P)H, the burst phase kinetics for the NADP+-dependent oxidation of 3alpha-diol is eliminated | Rattus norvegicus |
W227A | site-directed mutagenesis, highly reduced activity compared to the wild-type, very slow chemical transformation | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | transient-state and steady-state kinetics | Rattus norvegicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3alpha-androstanediol + NAD+ | Rattus norvegicus | steroid reduction direction is preferred | 5alpha-dihydrotestosterone + NADH + H+ | - |
r | |
5alpha-dihydrotestosterone + NADPH + H+ | Rattus norvegicus | steroid reduction direction is preferred | 3alpha-androstanediol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
isozyme AKR1C9 | - |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | isozyme AKR1C9 | Rattus norvegicus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2Z)-2-(5-hydroxy-4,6-dimethyl-2-oxo-1,2-dihydro-3H-indol-3-ylidene)-N,N-di(prop-2-en-1-yl)hydrazinecarbothioamide + NADPH + H+ | - |
Rattus norvegicus | ? | - |
r | |
3alpha-androstanediol + NAD+ | steroid reduction direction is preferred | Rattus norvegicus | 5alpha-dihydrotestosterone + NADH + H+ | - |
r | |
5alpha-dihydrotestosterone + NADPH + H+ | steroid reduction direction is preferred | Rattus norvegicus | 3alpha-androstanediol + NADP+ | - |
r | |
5alpha-dihydrotestosterone + NADPH + H+ | the chemical oxidation step is rate-limiting, W227 is very important for cataylsis, steroid reduction direction is preferred | Rattus norvegicus | 3alpha-androstanediol + NADP+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
3alpha-HSD | - |
Rattus norvegicus |
3alpha-hydroxysteroid dehydrogenase | - |
Rattus norvegicus |
AKR1C9 | - |
Rattus norvegicus |
More | see also EC 1.1.1.50, 3alpha-hydroxysteroid dehydrogenase (B-specific), for enzymes catalyzing the same reaction, but which are B-specific with respect to hydrogen transfer to/from NAD(P)+/NAD(P)H. Enzymes without information of stereospecificity are summarized under EC 1.1.1.50, but may in fact belong to EC 1.1.1.213 | Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | single turnover, and multiple burst and steady-state in turnover, kinetic isotope effects with 4-pro-R-[2H]-NADPH on recombinant wild-type and mutant enzymes | Rattus norvegicus | |
0.00063 | - |
(2Z)-2-(5-hydroxy-4,6-dimethyl-2-oxo-1,2-dihydro-3H-indol-3-ylidene)-N,N-di(prop-2-en-1-yl)hydrazinecarbothioamide | cofactor NADP+, pH 7.0, 25°C, steroid oxidation, recombinant mutant W227A | Rattus norvegicus | |
0.037 | - |
5alpha-dihydroprogesterone | cofactor NADPH, pH 7.0, 25°C, steroid reduction, recombinant mutant W227A | Rattus norvegicus | |
0.3 | - |
5alpha-dihydroprogesterone | cofactor NADPH, pH 7.0, 25°C, steroid reduction, recombinant wild-type enzyme | Rattus norvegicus | |
0.37 | - |
(2Z)-2-(5-hydroxy-4,6-dimethyl-2-oxo-1,2-dihydro-3H-indol-3-ylidene)-N,N-di(prop-2-en-1-yl)hydrazinecarbothioamide | cofactor NADP+, pH 7.0, 25°C, steroid oxidation, recombinant wild-type enzyme | Rattus norvegicus | |
0.4 | - |
5alpha-dihydroprogesterone | cofactor NADPH, pH 7.0, 25°C, steroid reduction, recombinant mutant R276M | Rattus norvegicus | |
0.43 | - |
5alpha-dihydroprogesterone | cofactor NADH, pH 7.0, 25°C, steroid reduction, recombinant wild-type enzyme | Rattus norvegicus | |
0.8 | - |
(2Z)-2-(5-hydroxy-4,6-dimethyl-2-oxo-1,2-dihydro-3H-indol-3-ylidene)-N,N-di(prop-2-en-1-yl)hydrazinecarbothioamide | cofactor NADP+, pH 7.0, 25°C, steroid oxidation, recombinant mutant R276M | Rattus norvegicus | |
0.85 | - |
(2Z)-2-(5-hydroxy-4,6-dimethyl-2-oxo-1,2-dihydro-3H-indol-3-ylidene)-N,N-di(prop-2-en-1-yl)hydrazinecarbothioamide | cofactor NAD+, pH 7.0, 25°C, steroid oxidation, recombinant wild-type enzyme | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | R276 plays an important role in cofactor binding | Rattus norvegicus | |
NAD+ | A-specific | Rattus norvegicus | |
NADH | A-specific | Rattus norvegicus | |
NADP+ | A-specific | Rattus norvegicus | |
NADPH | A-specific | Rattus norvegicus |