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Literature summary 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

EC Number Protein Variants Comment Organism
1.1.1.213 N167A site-directed mutagenesis, most impaired enzyme Rattus norvegicus
1.1.1.213 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
1.1.1.213 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
1.1.1.213 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
1.1.1.213 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.213 0.0011 0.0039 5alpha-androstan-3,17-dione NADPH Rattus norvegicus
1.1.1.213 0.0013 0.0441 3alpha-hydroxy-5alpha-androstan-17-one NAD+ Rattus norvegicus
1.1.1.213 0.0022 0.0081 NADPH
-
 Rattus norvegicus
1.1.1.213 0.0039 0.0304 5alpha-androstan-3,17-dione NADH Rattus norvegicus
1.1.1.213 0.004 0.821 NAD+
-
 Rattus norvegicus
1.1.1.213 0.0041 0.024 3alpha-hydroxy-5alpha-androstan-17-one NADP+ Rattus norvegicus
1.1.1.213 0.0052 0.1515 NADP+
-
 Rattus norvegicus
1.1.1.213 0.0057 0.0312 NADH
-
 Rattus norvegicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.213 37000
-
 1 * 37000, SDS-PAGE Rattus norvegicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.213 17beta-hydroxy-5alpha-androstan-3-one + NAD(P)H + H+ Rattus norvegicus
-
 3alpha,17beta-dihydroxy-5alpha-androstan + NAD(P)+
-
 ?
1.1.1.213 additional information Rattus norvegicus biosynthesis and inactivation of steroid hormones, physiological role is to inactivate circulating androgens, progestins and glucocorticoids ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.213 Rattus norvegicus P23457
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.213 anion exchange and Blue Sepharose column Rattus norvegicus
1.1.1.213 N167A chromatofocusing and gel filtration introduced between the to others chromatography steps Rattus norvegicus

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.213 a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ aldoketoreductase superfamily Rattus norvegicus
a
1.1.1.213 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
1.1.1.213 a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ ordered and sequential mechanism, nucleotide binds first and leaves last Rattus norvegicus
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.213 liver
-
 Rattus norvegicus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.213 0.08
-
 Q190A mutant, androsterone oxidation with NAD+ Rattus norvegicus
1.1.1.213 0.1
-
 S166A mutant, androsterone oxidation with NAD+ Rattus norvegicus
1.1.1.213 0.81
-
 Y216S mutant, androsterone oxidation with NADP+ Rattus norvegicus
1.1.1.213 1.55
-
 recombinant wild type enzyme, androsterone oxidation with NAD+ Rattus norvegicus

Storage Stability

EC Number Storage Stability Organism
1.1.1.213 -80°C Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.213 17beta-hydroxy-5alpha-androstan-3-one + NAD(P)H + H+
-
 Rattus norvegicus 3alpha,17beta-dihydroxy-5alpha-androstan + NAD(P)+
-
 ?
1.1.1.213 5alpha-androstan-3,17-dione + NAD(P)H
-
 Rattus norvegicus 3alpha-hydroxy-5alpha-androstan-17-one + NAD(P)+
-
 r
1.1.1.213 additional information biosynthesis and inactivation of steroid hormones, physiological role is to inactivate circulating androgens, progestins and glucocorticoids Rattus norvegicus ?
-
 ?

Subunits

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.213 25
-
 assay at Rattus norvegicus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.213 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
1.1.1.213 NAD+
-
 Rattus norvegicus
1.1.1.213 NADH
-
 Rattus norvegicus
1.1.1.213 NADP+
-
 Rattus norvegicus
1.1.1.213 NADPH
-
 Rattus norvegicus