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Literature summary for 1.1.1.213 extracted from

  • Penning, T.M.; Bennett, M.J.; Smith-Hoog, S.; Schlegel, B.P.; Jez, J.M.; Lewis, M.
    Structure and function of 3alpha-hydroxysteroid dehydrogenase (1997), Steroids, 62, 101-111.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
full-length cDNA gene is expressed in Escherichia coli DH5alpha Rattus norvegicus

Crystallization (Commentary)

Crystallization (Comment) Organism
vapor diffusion techniques using ammonium sulfate as precipitant Rattus norvegicus

Protein Variants

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

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 [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

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Rattus norvegicus 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
additional information non-metallo enzyme Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37029
-
1 * 37029 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 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

Organism UniProt Comment Textmining
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 first and leaves last 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
additional information
-
enzyme contains conserved catalytic tetrad of Asp50, Tyr55, Lys84 and His117 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+ 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

Subunits Comment Organism
monomer 1 * 37029 Rattus norvegicus
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
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

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