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Literature summary for 1.1.1.213 extracted from

  • Heredia, V.V.; Cooper, W.C.; Kruger, R.G.; Jin, Y.; Penning, T.M.
    Alanine scanning mutagenesis of the testosterone binding site of rat 3alpha-hydroxysteroid dehydrogenase demonstrates contact residues influence the rate-determining step (2004), Biochemistry, 43, 5832-5841.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutant enzymes in Escherichia coli C41(DE3) Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
F118A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, effects of mutation on binding constants and kinetics, overview Rattus norvegicus
F129A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, effects of mutation on binding constants and kinetics, overview Rattus norvegicus
L54A site-directed mutagenesis, reduced activity compared to the wild-type enzyme, effects of mutation on binding constants and kinetics, overview Rattus norvegicus
N306A site-directed mutagenesis, reduced activity compared to the wild-type enzyme, effects of mutation on binding constants and kinetics, overview Rattus norvegicus
T226A site-directed mutagenesis, reduced activity compared to the wild-type enzyme, effects of mutation on binding constants and kinetics, overview Rattus norvegicus
T24A site-directed mutagenesis, reduced activity compared to the wild-type enzyme, effects of mutation on binding constants and kinetics, overview Rattus norvegicus
W227A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, 23fold decreased affinity for progesterone, effects of mutation on binding constants and kinetics, overview Rattus norvegicus
Y310A site-directed mutagenesis, reduced activity compared to the wild-type enzyme, effects of mutation on binding constants and kinetics, overview Rattus norvegicus

Inhibitors

Inhibitors Comment Organism Structure
progesterone competitive to testosterone Rattus norvegicus
testosterone competitive to progesterone Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli C41(DE3), to homogeneity Rattus norvegicus

Reaction

Reaction Comment Organism Reaction ID
a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ ordered bi bi reaction mechanism Rattus norvegicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.006
-
purified recombinant mutant W227A, substrate androsterone Rattus norvegicus
0.01
-
purified recombinant mutant F118A, substrate androsterone Rattus norvegicus
0.03
-
purified recombinant mutant F129A, substrate androsterone Rattus norvegicus
0.08
-
purified recombinant mutant L54A, substrate androsterone Rattus norvegicus
0.09
-
purified recombinant mutant N306A, substrate androsterone Rattus norvegicus
0.1
-
purified recombinant mutant Y310A, substrate androsterone Rattus norvegicus
0.86
-
purified recombinant mutant T24A, substrate androsterone Rattus norvegicus
0.96
-
purified recombinant mutant T226A, substrate androsterone Rattus norvegicus
1.6
-
purified recombinant wild-type enzyme, substrate androsterone Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5beta-androstan-3alpha-ol-17-one + NAD(P)+
-
Rattus norvegicus 5beta-androstane-3,17-dione + NAD(P)H
-
r
5beta-pregnan-3alpha-ol-20-one + NAD(P)+
-
Rattus norvegicus 5beta-pregnane-3,20-dione + NAD(P)H
-
r
androsterone + NAD(P)+
-
Rattus norvegicus 5alpha-androstane-3,17-dione + NAD(P)H
-
r
progesterone + NAD(P)H
-
Rattus norvegicus 3alpha-hydroxy-pregn-4-ene-20-one + NAD(P)+
-
r
testosterone + NAD(P)H + H+
-
Rattus norvegicus 4-androsten-3alpha,17beta-diol + NAD(P)+
-
r

Synonyms

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
additional information kcat in steady-state, single turnover, burst in multiple turnover, steady-state in multiple turnover Rattus norvegicus
0.0053
-
androsterone steady-state, pH 8.0, 25°C, recombinant mutant W227A Rattus norvegicus
0.0083
-
androsterone steady-state, pH 8.0, 25°C, recombinant mutant F118A Rattus norvegicus
0.57
-
androsterone steady-state, pH 8.0, 25°C, recombinant mutant T226A Rattus norvegicus
0.83
-
androsterone steady-state, pH 8.0, 25°C, recombinant wild-type enzyme Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
NAD(P)+ A-specific Rattus norvegicus
NAD(P)H A-specific Rattus norvegicus