Cloned (Comment) | Organism |
---|---|
overexpression of GST-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00084 | - |
pregn-4-en-3,20-dione | pH 7.4, 37°C, recombinant wild-type enzyme | Homo sapiens | |
0.00213 | - |
pregn-4-en-3,20-dione | pH 7.4, 37°C, recombinant mutant V54L | Homo sapiens | |
0.00218 | - |
5alpha-dihydrotestosterone | pH 7.4, 37°C, recombinant wild-type enzyme | Homo sapiens | |
0.07103 | - |
5alpha-dihydrotestosterone | pH 7.4, 37°C, recombinant mutant V54L | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5alpha-dihydrotestosterone + NADPH + H+ | Homo sapiens | - |
5alpha-androstane-3alpha,17beta-diol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q04828 | - |
- |
Purification (Comment) | Organism |
---|---|
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5alpha-dihydrotestosterone + NADPH + H+ | - |
Homo sapiens | 5alpha-androstane-3alpha,17beta-diol + NADP+ | - |
r | |
pregn-4-en-3,20-dione + NADPH + H+ | - |
Homo sapiens | 3alpha-hydroxypregn-4-en-20-one + NADP+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
3-alpha hydroxysteroid dehydrogenase type 3 | - |
Homo sapiens |
3alpha-HSD3 | - |
Homo sapiens |
AKR1C1 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0035 | - |
pregn-4-en-3,20-dione | pH 7.4, 37°C, recombinant wild-type enzyme | Homo sapiens | |
0.0392 | - |
pregn-4-en-3,20-dione | pH 7.4, 37°C, recombinant mutant V54L | Homo sapiens | |
0.0612 | - |
5alpha-dihydrotestosterone | pH 7.4, 37°C, recombinant wild-type enzyme | Homo sapiens | |
0.1153 | - |
5alpha-dihydrotestosterone | pH 7.4, 37°C, recombinant mutant V54L | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | binding structure and mechanism, overview | Homo sapiens | |
NADPH | - |
Homo sapiens |
General Information | Comment | Organism |
---|---|---|
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 |
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 |
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 Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.623 | - |
5alpha-dihydrotestosterone | pH 7.4, 37°C, recombinant mutant V54L | Homo sapiens | |
4.167 | - |
pregn-4-en-3,20-dione | pH 7.4, 37°C, recombinant wild-type enzyme | Homo sapiens | |
18.4 | - |
pregn-4-en-3,20-dione | pH 7.4, 37°C, recombinant mutant V54L | Homo sapiens | |
28.07 | - |
5alpha-dihydrotestosterone | pH 7.4, 37°C, recombinant wild-type enzyme | Homo sapiens |