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Literature summary extracted from

  • Zhang, B.; Zhu, D.W.; Hu, X.J.; Zhou, M.; Shang, P.; Lin, S.X.
    Human 3-alpha hydroxysteroid dehydrogenase type 3 (3alpha-HSD3) the V54L mutation restricting the steroid alternative binding and enhancing the 20?-HSD activity (2014), J. Steroid Biochem. Mol. Biol., 141, 135-143 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.357 overexpression of GST-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.357 3alpha-HSD3-NADP+-progesterone complex and 3alpha-HSD3 mutant V54L-NADP+-progesterone complex, sitting drop vapor diffusion method, room temperature, the reservoir solution contains 100 mM sodium cacodylate, pH 6.0, 200 mM ammonium sulfate, 24-26% w/v PEG 3350, with 0.8 mM progesterone, 3-14 days, X-ray diffraction structure determination and analysis at 2.0-2.2 A resolution, molecular replacement with search model, PDB ID 1J96. Progesterone adopts two different binding modes to form complexes within the wild-type enzyme, with one binding mode similar to the orientation of a bile acid (ursodeoxycholate) in the reported ternary complex of human 3alpha-HSD3-NADP+-ursodeoxycholate and the other binding mode resembling the orientation of 20alpha-OHProg in the ternary complex of human 20alpha-HSD-NADP+-20alpha-OHProg Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.357 V54L site-directed mutagenesis, the change renders the sequence identical to that of human 20-alpha hydroxysteroid dehydrogenase. The V54L mutation directly restricts the steroid binding modes to a unique one, which resembles the orientation of 20alpha-OHProg within human 20alpha-HSD. The kinetic study shows that the V54L mutation significantly decreases the 3alpha-HSD activity for the reduction of 5alpha-dihydrotestosterone, while this mutation enhances the 20alpha-HSD activity to convert progesterone Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.357 0.00084
-
 pregn-4-en-3,20-dione pH 7.4, 37°C, recombinant wild-type enzyme Homo sapiens
1.1.1.357 0.00213
-
 pregn-4-en-3,20-dione pH 7.4, 37°C, recombinant mutant V54L Homo sapiens
1.1.1.357 0.00218
-
 5alpha-dihydrotestosterone pH 7.4, 37°C, recombinant wild-type enzyme Homo sapiens
1.1.1.357 0.07103
-
 5alpha-dihydrotestosterone pH 7.4, 37°C, recombinant mutant V54L Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.357 5alpha-dihydrotestosterone + NADPH + H+ Homo sapiens
-
 5alpha-androstane-3alpha,17beta-diol + NADP+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.357 Homo sapiens Q04828
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.357 recombinant GST-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by glutathione affinity chromatography, cleavage of the tag by thrombin, and anion exchange chromatography Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.357 5alpha-dihydrotestosterone + NADPH + H+
-
 Homo sapiens 5alpha-androstane-3alpha,17beta-diol + NADP+
-
 r
1.1.1.357 pregn-4-en-3,20-dione + NADPH + H+
-
 Homo sapiens 3alpha-hydroxypregn-4-en-20-one + NADP+
-
 r

Synonyms

EC Number Synonyms Comment Organism
1.1.1.357 3-alpha hydroxysteroid dehydrogenase type 3
-
 Homo sapiens
1.1.1.357 3alpha-HSD3
-
 Homo sapiens
1.1.1.357 AKR1C1
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.357 37
-
 assay at Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.357 0.0035
-
 pregn-4-en-3,20-dione pH 7.4, 37°C, recombinant wild-type enzyme Homo sapiens
1.1.1.357 0.0392
-
 pregn-4-en-3,20-dione pH 7.4, 37°C, recombinant mutant V54L Homo sapiens
1.1.1.357 0.0612
-
 5alpha-dihydrotestosterone pH 7.4, 37°C, recombinant wild-type enzyme Homo sapiens
1.1.1.357 0.1153
-
 5alpha-dihydrotestosterone pH 7.4, 37°C, recombinant mutant V54L Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.357 7.4
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.357 NADP+ binding structure and mechanism, overview Homo sapiens
1.1.1.357 NADPH
-
 Homo sapiens

General Information

EC Number General Information Comment Organism
1.1.1.357 evolution human 3alpha-HSD3 shares 97.8% sequence identity with human 20-alpha hydroxysteroid dehydrogenase (20alpha-HSD) and there is only one amino acid difference (residue 54) that is located in their steroid binding pockets. 20alpha-HSD displays a distinctive ability in transforming progesterone to 20alpha-hydroxy-progesterone Homo sapiens
1.1.1.357 malfunction the V54L mutation significantly decreases the 3alpha-HSD activity for the reduction of 5alpha-dihydrotestosterone, while this mutation enhances the 20alpha-HSD activity to convert progesterone Homo sapiens
1.1.1.357 physiological function 3-alpha hydroxysteroid dehydrogenase type 3 has an essential role in the inactivation of 5alpha-dihydrotestosterone preventing binding and activation of androgen receptor from overflowing androgen Homo sapiens

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.357 1.623
-
 5alpha-dihydrotestosterone pH 7.4, 37°C, recombinant mutant V54L Homo sapiens
1.1.1.357 4.167
-
 pregn-4-en-3,20-dione pH 7.4, 37°C, recombinant wild-type enzyme Homo sapiens
1.1.1.357 18.4
-
 pregn-4-en-3,20-dione pH 7.4, 37°C, recombinant mutant V54L Homo sapiens
1.1.1.357 28.07
-
 5alpha-dihydrotestosterone pH 7.4, 37°C, recombinant wild-type enzyme Homo sapiens