Literature summary for 1.1.1.213 extracted from

Couture, J.F.; de Jesus-Tran, K.P.; Roy, A.M.; Cantin, L.; Cote, P.L.; Legrand, P.; Luu-The, V.; Labrie, F.; Breton, R.
Comparison of crystal structures of human type 3 3alpha-hydroxysteroid dehydrogenase reveals an 'induced-fit' mechanism and a conserved basic motif involved in the binding of androgen (2005), Protein Sci., 14, 1485-1497.

Search for more literature for 1.1.1.213

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of GST-fusion wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Homo sapiens

Crystallization (Commentary)

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

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

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 [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
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

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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Homo sapiens	P52895	-	-

Purification (Commentary)

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

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	a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	-	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

Subunits	Comment	Organism
dimer	asymmetric heterodimer, crystal structure analysis	Homo sapiens

Synonyms

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 [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
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

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	cofactor binding site structure	Homo sapiens
NADPH	cofactor binding site structure	Homo sapiens