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Literature summary for 1.1.1.213 extracted from

  • Ma, H.; Ratnam, K.; Penning, T.M.
    Mutation of nicotinamide pocket residues in rat liver 3alpha-hydroxysteroid dehydrogenase reveals different modes of cofactor binding (2000), Biochemistry, 39, 102-109.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
N167A site-directed mutagenesis, most impaired enzyme Rattus norvegicus
Q190A site-directed mutagenesis, decreased binding affinity to NADP(H), only binding of cofactor is affected, residue is located at the catalytic cente Rattus norvegicus
R276M elimination of salt bridge between Arg276 and the 2'-phosphate of AMP results in 100fold decrease of the affinity to NADP(H) Rattus norvegicus
S166A site-directed mutagenesis, decreased binding affinity to NADP(H), only binding of cofactor is affected, residue is located at the catalytic center Rattus norvegicus
Y216S site-directed mutagenesis, decreased binding affinity to NADP(H), only binding of cofactor is affected, residue is located at the catalytic cente Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0011 0.0039 5alpha-androstan-3,17-dione NADPH Rattus norvegicus
0.0013 0.0441 3alpha-hydroxy-5alpha-androstan-17-one NAD+ Rattus norvegicus
0.0022 0.0081 NADPH
-
Rattus norvegicus
0.0039 0.0304 5alpha-androstan-3,17-dione NADH Rattus norvegicus
0.004 0.821 NAD+
-
Rattus norvegicus
0.0041 0.024 3alpha-hydroxy-5alpha-androstan-17-one NADP+ Rattus norvegicus
0.0052 0.1515 NADP+
-
Rattus norvegicus
0.0057 0.0312 NADH
-
Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37000
-
1 * 37000, SDS-PAGE 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 biosynthesis and inactivation of steroid hormones, physiological role is to inactivate circulating androgens, progestins and glucocorticoids ?
-
?

Organism

Organism UniProt Comment Textmining
Rattus norvegicus P23457
-
-

Purification (Commentary)

Purification (Comment) Organism
anion exchange and Blue Sepharose column Rattus norvegicus
N167A chromatofocusing and gel filtration introduced between the to others chromatography steps 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
a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ ordered and sequential mechanism, 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
0.08
-
Q190A mutant, androsterone oxidation with NAD+ Rattus norvegicus
0.1
-
S166A mutant, androsterone oxidation with NAD+ Rattus norvegicus
0.81
-
Y216S mutant, androsterone oxidation with NADP+ Rattus norvegicus
1.55
-
recombinant wild type enzyme, androsterone oxidation with NAD+ Rattus norvegicus

Storage Stability

Storage Stability Organism
-80°C 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)+
-
?
5alpha-androstan-3,17-dione + NAD(P)H
-
Rattus norvegicus 3alpha-hydroxy-5alpha-androstan-17-one + NAD(P)+
-
r
additional information biosynthesis and inactivation of steroid hormones, physiological role is to inactivate circulating androgens, progestins and glucocorticoids Rattus norvegicus ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 37000, SDS-PAGE Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
additional information transfer of the 4-pro-R hydrogen from A-face of cofactor to the beta-face of the steroid substrate. Different binding modes for NAD(H) and NADP(H) Rattus norvegicus
NAD+
-
Rattus norvegicus
NADH
-
Rattus norvegicus
NADP+
-
Rattus norvegicus
NADPH
-
Rattus norvegicus