Cloned (Comment) | Organism |
---|---|
overexpression of GST-fusion wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type enzyme, hanging drop vapour diffusion method, 4°C, mixing of equal volumes of 14 mg/ml protein solution and mother liquor containing 0.1 M sodium citrate, pH 6.5, 0.1 M ammonium acetate, and 24-30% PEG 4000, cryoprotection of crystals in 20% ethylene glycol in mother liquor, X-ray diffraction structure determination and analysis at 1.9 A resolution, molecular replacement study | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
R301L | site-directed mutagenesis, the mutation greatly affects the 3alpha-hydroxysteroid dehydrogenase activity towards 5alpha-dihydrotestosterone and almost completely abolishes the 17beta-hydroxysteroid dehydrogenase activity of the enzyme | Homo sapiens |
R304L | site-directed mutagenesis, the mutation greatly affects the 3alpha-hydroxysteroid dehydrogenase activity towards 5alpha-dihydrotestosterone and abolishes the 17beta-hydroxysteroid dehydrogenase activity of the enzyme | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0057 | - |
5alpha-dihydrotestosterone | pH 7.5, recombinant wild-type enzyme | Homo sapiens | |
0.0059 | - |
5alpha-dihydrotestosterone | pH 7.5, recombinant mutant R301L | Homo sapiens | |
0.012 | - |
5alpha-dihydrotestosterone | pH 7.5, recombinant mutant R304L | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
citrate | bound to the steroid-binding cavity via Tyr55 and His117, involved in the induced fit mechanism | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5alpha-dihydrotestosterone + NADPH | Homo sapiens | the enzyme plays a crucial role in the regulation of the intracellular concentrations of testosterone and 5alpha-dihydrotestosterone, two steroids directly linked to the etiology and the progression of many prostate diseases and cancer, overview | 5alpha-androstane-3alpha,17beta-diol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P52895 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-fusion wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by glutathione affinity and anion exchange chromatography, and gel filtration to homogeneity | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ | the enzyme reveals an 'induced-fit' mechanism involving a bound citrate ion and a conserved basic motif involved in the binding of androgen, the enzyme also binds many structurally different molecules such as 4-hydroxynonenal, polycyclic aromatic hydrocarbons, and indanone, modeling of substrate binding | Homo sapiens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-indanone + NADPH | - |
Homo sapiens | ? | - |
? | |
4-hydroxynonenal + NADPH | - |
Homo sapiens | ? | - |
? | |
5alpha-dihydrotestosterone + NADPH | the enzyme plays a crucial role in the regulation of the intracellular concentrations of testosterone and 5alpha-dihydrotestosterone, two steroids directly linked to the etiology and the progression of many prostate diseases and cancer, overview | Homo sapiens | 5alpha-androstane-3alpha,17beta-diol + NADP+ | - |
r | |
5alpha-dihydrotestosterone + NADPH | i.e. DHT, substrate and cofactor binding site structure | Homo sapiens | 5alpha-androstane-3alpha,17beta-diol + NADP+ | - |
r | |
additional information | besides 3alpha-hydroxysteroid dehydrogenase activity, the enzyme shows also 17beta-hydroxysteroid dehydrogenase activity towards 5alpha-dihydrotestosterone, EC 1.1.1.239, overview, the enzyme reveals an 'induced-fit' mechanism and a conserved basic motif involved in the binding of androgen, the enzyme also binds many structurally different molecules such as 4-hydroxynonenal, polycyclic aromatic hydrocarbons, and indanone | Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | asymmetric heterodimer, crystal structure analysis | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
3alpha-HSD3 | - |
Homo sapiens |
AKR1C2 | - |
Homo sapiens |
More | cf. EC 1.1.1.239, the enzyme belongs to the AKR1C subfamily | Homo sapiens |
type 3 3alpha-hydroxysteroid dehydrogenase | - |
Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.9 | - |
5alpha-dihydrotestosterone | pH 7.5, recombinant mutant R304L | Homo sapiens | |
22.1 | - |
5alpha-dihydrotestosterone | pH 7.5, recombinant mutant R301L | Homo sapiens | |
1339 | - |
5alpha-dihydrotestosterone | pH 7.5, recombinant wild-type enzyme | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | cofactor binding site structure | Homo sapiens | |
NADPH | cofactor binding site structure | Homo sapiens |