Cloned (Comment) | Organism |
---|---|
full-length cDNA gene is expressed in Escherichia coli DH5alpha | Rattus norvegicus |
Crystallization (Comment) | Organism |
---|---|
vapor diffusion techniques using ammonium sulfate as precipitant | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
C217A | resistant to inactivation by secosteroids, therefore Cys217 is the point of covalent attachment of acetylenic ketones | Rattus norvegicus |
D50E | 1/30th catalytic efficiency of wild type, unlikely to be the general amino acid for catalysis | Rattus norvegicus |
D50N | 1/30th catalytic efficiency of wild type, unlikely to be the general amino acid for catalysis | Rattus norvegicus |
H117A | 1/500th catalytic efficiency of wild type, unlikely to be the general amino acid for catalysis | Rattus norvegicus |
K84M | inactive, unable to bind steroids | Rattus norvegicus |
K84R | inactive, unable to bind steroids | Rattus norvegicus |
additional information | positions of Tyr/Lys pair are conserved across the aldo-keto reductase and short-chain dehydrogenase/reductase family. Tyr retains ability to form ternary complex and acts as general acid | Rattus norvegicus |
W148Y | can catalyse steroid oxidoreduction similar to wild type, plays no role in steroid binding or catalysis | Rattus norvegicus |
W227Y | mainly influenced in steroid binding | Rattus norvegicus |
W86Y | plays role in cofactor and steroid binding | Rattus norvegicus |
Y205F | kinetically indistinguishable from the wild type, no general amino acid for catalysis in 3alpha-hydroxysteroid dehydrogenase | Rattus norvegicus |
Y55F | inactive, unable to perform steroid oxidoreduction, strongest candidate for the general amino acid | Rattus norvegicus |
Y55S | inactive, strongest candidate for the general amino acid | Rattus norvegicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acetylenic ketones | inactivation by forming Michael adducts with enzyme nucleophiles | Rattus norvegicus | |
cacodylate | - |
Rattus norvegicus | |
citrate | - |
Rattus norvegicus | |
D-glucose 6-phosphate | - |
Rattus norvegicus | |
zopolrestat | - |
Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0012 | - |
17beta-hydroxy-5alpha-androstan-3-one | cloned human liver enzyme type I | Rattus norvegicus | |
0.0041 | - |
5alpha-androstan-3,17-dione | recombinant wild type enzyme | Rattus norvegicus | |
0.0042 | - |
5alpha-androstan-3,17-dione | Y205F | Rattus norvegicus | |
0.0192 | - |
17beta-hydroxy-5alpha-androstan-3-one | cloned human liver enzyme type II | Rattus norvegicus | |
0.027 | - |
NADH | recombinant wild type enzyme | Rattus norvegicus | |
0.029 | - |
3alpha-hydroxy-5alpha-androstan-17-one | Y205F | Rattus norvegicus | |
0.03 | - |
NADH | Y205F | Rattus norvegicus | |
0.042 | 0.408 | 3alpha-hydroxy-5alpha-androstan-17-one | - |
Rattus norvegicus | |
0.046 | - |
3alpha-hydroxy-5alpha-androstan-17-one | recombinant wild type enzyme | Rattus norvegicus | |
0.83 | - |
NAD+ | Y205F | Rattus norvegicus | |
0.94 | - |
NAD+ | recombinant wild type enzyme | Rattus norvegicus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Rattus norvegicus | 5829 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | non-metallo enzyme | Rattus norvegicus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
37029 | - |
1 * 37029 | Rattus norvegicus |
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 | regulation of the amount of androgen in prostate, high levels of substrate are required for normal and abnormal growth of prostate | 3alpha,17beta-dihydroxy-5alpha-androstan + NAD(P)+ | - |
r | |
5alpha-pregnan-3,20-dione + NAD(P)H + H+ | Rattus norvegicus | regulation of the amount of allosteric agonists that can bind to the GABA receptor in brain | 5alpha-pregnan-3alpha-ol-20-one + NAD(P)+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
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 first and leaves last | Rattus norvegicus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Rattus norvegicus | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
enzyme contains conserved catalytic tetrad of Asp50, Tyr55, Lys84 and His117 | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
17beta-hydroxy-5alpha-androstan-3-one + NAD(P)H + H+ | regulation of the amount of androgen in prostate, high levels of substrate are required for normal and abnormal growth of prostate | Rattus norvegicus | 3alpha,17beta-dihydroxy-5alpha-androstan + NAD(P)+ | - |
r | |
3alpha-hydroxy-5alpha-androstan-17-one + NAD+ | - |
Rattus norvegicus | 5alpha-androstan-3,17-dione + NADH | - |
r | |
5alpha-pregnan-3,20-dione + NAD(P)H + H+ | regulation of the amount of allosteric agonists that can bind to the GABA receptor in brain | Rattus norvegicus | 5alpha-pregnan-3alpha-ol-20-one + NAD(P)+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 37029 | Rattus norvegicus |
More | (alpha/beta)8-barrel fold | Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.267 | - |
5alpha-androstan-3,17-dione | Y205F | Rattus norvegicus | |
0.283 | - |
NADH | recombinant wild type enzyme | Rattus norvegicus | |
0.283 | - |
5alpha-androstan-3,17-dione | recombinant wild type enzyme | Rattus norvegicus | |
0.367 | - |
NADH | Y205F | Rattus norvegicus | |
1.13 | - |
3alpha-hydroxy-5alpha-androstan-17-one | Y205F | Rattus norvegicus | |
1.13 | - |
NAD+ | recombinant wild type enzyme | Rattus norvegicus | |
1.25 | - |
3alpha-hydroxy-5alpha-androstan-17-one | recombinant wild type enzyme | Rattus norvegicus | |
1.43 | - |
NAD+ | Y205F | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | 4-pro-(R) hydrogen is transferred from the A-face of the cofactor to the B-face of the steroid | Rattus norvegicus | |
NAD+ | - |
Rattus norvegicus | |
NADH | - |
Rattus norvegicus | |
NADP+ | - |
Rattus norvegicus | |
NADPH | preferred | Rattus norvegicus |