Information on EC 1.1.1.357 - 3alpha-hydroxysteroid 3-dehydrogenase and Organism(s) Homo sapiens

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Homo sapiens


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota


The taxonomic range for the selected organisms is: Homo sapiens

EC NUMBER
COMMENTARY hide
1.1.1.357
-
RECOMMENDED NAME
GeneOntology No.
3alpha-hydroxysteroid 3-dehydrogenase
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
brassinosteroid biosynthesis I
-
-
brassinosteroid biosynthesis II
-
-
SYSTEMATIC NAME
IUBMB Comments
3alpha-hydroxysteroid:NAD(P)+ 3-oxidoreductase
The enzyme acts on multiple 3alpha-hydroxysteroids, such as androsterone and 5 alpha-dihydrotestosterone. The mammalian enzymes are involved in inactivation of steroid hormones, while the bacterial enzymes are involved in steroid degradation. This entry stands for enzymes whose stereo-specificity with respect to NAD+ or NADP+ is not known. [cf. EC 1.1.1.50, 3alpha-hydroxysteroid 3-dehydrogenase (Si-specific) and EC 1.1.1.213, 3alpha-hydroxysteroid 3-dehydrogenase (Re-specific)].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-tetralol + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
(S)-1,2,3,4-tetrahydronaphth-1-ol + NADP+
1,2,3,4-tetrahydronaphth-1-one + NADPH + H+
show the reaction diagram
-
-
-
-
?
(S)-indan-1-ol + NADP+
indan-1-one + NADPH + H+
show the reaction diagram
-
-
-
-
?
(S)-tetralol + NADP+
1-tetralone + NADPH + H+
show the reaction diagram
-
-
-
-
?
1-acenaphthenol + NADP+
acenaphthylen-1(2H)-one + NADPH + H+
show the reaction diagram
-
-
-
?
17beta-estradiol + NAD(P)H + H+
estrone + NAD(P)+
show the reaction diagram
-
-
-
-
?
20alpha-hydroxyprogesterone + NAD(P)H + H+
progesterone + NAD(P)+
show the reaction diagram
-
-
-
-
?
3alpha-androstanediol + NAD(P)+
5alpha-dihydrotestosterone + NAD(P)H + H+
show the reaction diagram
-
-
-
-
r
3alpha-androstanediol + NAD+
5alpha-dihydrotestosterone + NADH + H+
show the reaction diagram
-
-
-
-
?
5alpha-androstan-3alpha-ol-17-one + NADP+
(5alpha)-androstan-3,17-dione + NADPH + H+
show the reaction diagram
-
-
-
-
?
5alpha-androstane-3,17-dione + NAD(P)H + H+
androsterone + NAD(P)+
show the reaction diagram
-
-
-
-
?
5alpha-androstane-3alpha,17beta-diol + NADP+
(5alpha)-androstan-17-beta-ol-3-one + NADPH + H+
show the reaction diagram
-
-
-
-
?
5alpha-dihydroprogesterone + NADPH + H+
allopregnanolone + 5alpha,20alpha-tetrahydroprogesterone + NADP+
show the reaction diagram
-
-
-
r
5alpha-dihydrotestosterone + NAD(P)H + H+
3alpha-androstanediol + NAD(P)+
show the reaction diagram
5alpha-dihydrotestosterone + NADPH + H+
3alpha-androstanediol + NADP+
show the reaction diagram
5alpha-dihydrotestosterone + NADPH + H+
5alpha-androstane-3alpha,17beta-diol + NADP+
show the reaction diagram
5beta-androstan-3alpha-ol-17-one + NADP+
(5beta)-androstan-3,17-dione + NADPH + H+
show the reaction diagram
-
-
-
-
?
5beta-androstane-3alpha,17beta-diol + NADP+
5beta-androstan-17beta-ol-3-one + NADPH + H+
show the reaction diagram
-
-
-
-
?
5beta-dihydroprogesterone + NADPH + H+
pregnanolone + NADP+
show the reaction diagram
-
-
-
r
5beta-pregnane-3alpha,20alpha-diol + NADP+
(5beta)-pregnan-20alpha-ol-3-one + NADPH + H+
show the reaction diagram
-
-
-
-
?
7alpha,12alpha-dihydroxy-5beta-cholestan-3-one + NAD(P)+
3alpha,7alpha,12alpha-trihydroxy-5beta-cholestane + NAD(P)H + H+
show the reaction diagram
-
-
-
-
?
9alpha,11beta-prostaglandin F2 + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
a 3-alpha-hydroxysteroid + NAD(P)+
a 3-oxosteroid + NAD(P)H + H+
show the reaction diagram
-
-
-
?
a 3alpha-hydroxysteroid + NAD(P)+
a 3-oxosteroid + NAD(P)H + H+
show the reaction diagram
-
-
-
-
r
allopregnanolone + 5alpha,20alpha-tetrahydroprogesterone + NADP+
5alpha-dihydroprogesterone + NADPH + H+
show the reaction diagram
-
-
-
r
androsterone + NAD(P)+
5alpha-androstane-3,17-dione + NAD(P)H + H+
show the reaction diagram
-
-
-
-
?
androsterone + NAD+
(5alpha)-androstane-3,17-dione + NADH + H+
show the reaction diagram
-
-
-
-
?
androsterone + NADP+
(5alpha)-androstane-3,17-dione + NADPH + H+
show the reaction diagram
-
-
-
-
?
androsterone + NADP+
5alpha-androstane-3,17-dione + NADPH + H+
show the reaction diagram
-
-
-
?
chenodeoxycholic acid + NADP+
(5beta,7alpha,8xi)-7-hydroxy-3-oxocholan-24-oic acid + NADPH + H+
show the reaction diagram
-
-
-
?
chenodeoxycholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
cholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
deoxycholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
dihydrotestosterone + NAD(P)H + H+
5alpha-androstane-3alpha,17beta-diol + NAD(P)+
show the reaction diagram
-
-
-
-
?
estrone + NAD(P)H + H+
17beta-estradiol + NAD(P)+
show the reaction diagram
-
-
-
-
?
lithocholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
pregn-4-en-3,20-dione + NADPH + H+
3alpha-hydroxypregn-4-en-20-one + NADP+
show the reaction diagram
-
-
-
r
progesterone + NAD(P)H + H+
20alpha-hydroxyprogesterone + NAD(P)+
show the reaction diagram
-
-
-
-
?
testosterone + NAD(P)H + H+
DELTA4-androstene-3,17-dione + NAD(P)+
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(R)-tetralol + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
(S)-indan-1-ol + NADP+
indan-1-one + NADPH + H+
show the reaction diagram
-
-
-
-
?
(S)-tetralol + NADP+
1-tetralone + NADPH + H+
show the reaction diagram
-
-
-
-
?
17beta-estradiol + NAD(P)H + H+
estrone + NAD(P)+
show the reaction diagram
-
-
-
-
?
20alpha-hydroxyprogesterone + NAD(P)H + H+
progesterone + NAD(P)+
show the reaction diagram
-
-
-
-
?
3alpha-androstanediol + NAD(P)+
5alpha-dihydrotestosterone + NAD(P)H + H+
show the reaction diagram
-
-
-
-
r
5alpha-androstane-3,17-dione + NAD(P)H + H+
androsterone + NAD(P)+
show the reaction diagram
-
-
-
-
?
5alpha-androstane-3alpha,17beta-diol + NADP+
(5alpha)-androstan-17-beta-ol-3-one + NADPH + H+
show the reaction diagram
-
-
-
-
?
5alpha-dihydroprogesterone + NADPH + H+
allopregnanolone + 5alpha,20alpha-tetrahydroprogesterone + NADP+
show the reaction diagram
P52895
-
-
-
r
5alpha-dihydrotestosterone + NAD(P)H + H+
3alpha-androstanediol + NAD(P)+
show the reaction diagram
5alpha-dihydrotestosterone + NADPH + H+
3alpha-androstanediol + NADP+
show the reaction diagram
-
physiological inactivation of the most potent androgen 5alpha dihydrotestosterone
-
-
r
5alpha-dihydrotestosterone + NADPH + H+
5alpha-androstane-3alpha,17beta-diol + NADP+
show the reaction diagram
Q04828
-
-
-
r
5beta-androstan-3alpha-ol-17-one + NADP+
(5beta)-androstan-3,17-dione + NADPH + H+
show the reaction diagram
-
-
-
-
?
5beta-androstane-3alpha,17beta-diol + NADP+
5beta-androstan-17beta-ol-3-one + NADPH + H+
show the reaction diagram
-
-
-
-
?
5beta-dihydroprogesterone + NADPH + H+
pregnanolone + NADP+
show the reaction diagram
P52895
-
-
-
r
7alpha,12alpha-dihydroxy-5beta-cholestan-3-one + NAD(P)+
3alpha,7alpha,12alpha-trihydroxy-5beta-cholestane + NAD(P)H + H+
show the reaction diagram
-
-
-
-
?
a 3alpha-hydroxysteroid + NAD(P)+
a 3-oxosteroid + NAD(P)H + H+
show the reaction diagram
-
-
-
-
r
allopregnanolone + 5alpha,20alpha-tetrahydroprogesterone + NADP+
5alpha-dihydroprogesterone + NADPH + H+
show the reaction diagram
P52895
-
-
-
r
androsterone + NAD(P)+
5alpha-androstane-3,17-dione + NAD(P)H + H+
show the reaction diagram
-
-
-
-
?
androsterone + NAD+
(5alpha)-androstane-3,17-dione + NADH + H+
show the reaction diagram
-
-
-
-
?
androsterone + NADP+
(5alpha)-androstane-3,17-dione + NADPH + H+
show the reaction diagram
-
-
-
-
?
chenodeoxycholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
cholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
deoxycholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
estrone + NAD(P)H + H+
17beta-estradiol + NAD(P)+
show the reaction diagram
-
-
-
-
?
lithocholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
-
?
progesterone + NAD(P)H + H+
20alpha-hydroxyprogesterone + NAD(P)+
show the reaction diagram
-
-
-
-
?
testosterone + NAD(P)H + H+
DELTA4-androstene-3,17-dione + NAD(P)+
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
isoform AKR1C2 functions preferentially as a 3-ketosteroid reductase
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
NADPH
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(3,6-dihydropyridin-1(2H)-yl)(1H-indol-2-yl)methanone
-
crystal structure analysis of enzyme-inhibitor complex
(5-methyl-1H-indol-2-yl)(4-propylpiperidin-1-yl)methanone
-
crystal structure analysis of enzyme-inhibitor complex
17beta-estradiol
-
competitive inhibitor
3,5,3',5'-tetraiodothyropropionic acid
-
competitive inhibitor
3-pentyl-2-[[(pyridin-2-yl)methyl]sulfanyl]-7-(pyrrolidine-1-carbonyl)quinazolin-4(3H)-one
-
crystal structure analysis of enzyme-inhibitor complex
4-chloro-N-[(4-chlorophenyl)methyl]-5-nitro-1H-pyrazole-3-carboxamide
-
crystal structure analysis of enzyme-inhibitor complex
4-nitro-2-([4-[3-(trifluoromethyl)phenyl]piperazin-1-yl]methyl)phenol
-
crystal structure analysis of enzyme-inhibitor complex
5-(benzenesulfonyl)-2-nitrophenol
-
crystal structure analysis of enzyme-inhibitor complex
chenodeoxycholic acid
-
-
Flufenamic acid
Hexestrol
-
-
Ibuprofen
-
-
indomethacin
-
-
lithocholic acid
-
-
Medroxyprogesterone acetate
-
-
Phenolphthalein
ursodeoxycholate
-
micromolar inhibitor, competitive inhibitor versus 5alpha-dihydroprogesterone and uncompetitive inhibitor versus NADPH
Ursodeoxycholic acid
-
-
additional information
-
crystal structures of complexes of 17beta-HSD5 with structurally diverse inhibitors derived from high-throughput inhibitor screening, overview. Analysis of interactions between 17beta-HSD5 and inhibitors at the atomic level which enable structure-based drug design for anti-CRPC therapy
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(R)-ibuprofen
-
167% activity at 1 mM
Clofibric acid
-
240% activity at 0.1 mM
D-thyroxine
-
260% activity at 1 mM
L-thyroxine
-
237% activity at 1 mM
Sulfobromophthalein
-
328% activity at 0.005 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.16 - 0.68
(R)-tetralol
0.14 - 0.26
(S)-1,2,3,4-tetrahydronaphth-1-ol
0.146 - 0.52
(S)-indan-1-ol
0.11 - 0.29
(S)-tetralol
0.0105
3alpha-androstanediol
-
isoform AKR1C4, at pH 7.4 and 25C
0.002 - 0.0022
5alpha-Androstan-3alpha-ol-17-one
0.00144
5alpha-androstane-3,17-dione
-
isoform AKR1C4, at pH 7.4 and 25C
0.0008 - 0.0031
5alpha-androstane-3alpha,17beta-diol
0.0018
5alpha-dihydroprogesterone
-
in 100 mM potassium phosphate, pH 7.0, at 25C
0.0012 - 0.07103
5alpha-dihydrotestosterone
0.0009 - 0.0018
5beta-Androstan-3alpha-ol-17-one
0.0012 - 0.0022
5beta-androstane-3alpha,17beta-diol
0.001
5beta-pregnane-3alpha,20alpha-diol
-
native enzyme, at pH 7.4 and 25C; recombinant enzyme, at pH 7.4 and 25C
0.208 - 0.22
9alpha,11beta-prostaglandin F2
0.0015
allopregnanolone
-
in 100 mM potassium phosphate, pH 7.0, at 25C
0.0005 - 0.00504
androsterone
0.0011 - 0.0029
chenodeoxycholic acid
0.032 - 0.059
cholic acid
0.0013 - 0.0097
deoxycholic acid
0.0011
dihydrotestosterone
-
pH 7.5, 37C
0.001 - 0.0019
lithocholic acid
1 - 1.2
NAD+
0.00023 - 0.0017
NADP+
0.00007
NADPH
-
in 100 mM potassium phosphate, pH 7.0, at 25C
0.00084 - 0.00213
pregn-4-en-3,20-dione
additional information
additional information
-
Km-value for the 17beta-oxidation of testosterone is 0.00067 mM, and Km-value for the 17beta-reduction of androstenedione is 0.00138 mM
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.18 - 0.77
(R)-tetralol
0.2 - 0.27
(S)-1,2,3,4-tetrahydronaphth-1-ol
0.1 - 0.4
(S)-indan-1-ol
0.2 - 0.43
(S)-tetralol
0.035
3alpha-androstanediol
-
isoform AKR1C4, at pH 7.4 and 25C
0.007
5alpha-Androstan-3alpha-ol-17-one
-
native enzyme, at pH 7.4 and 25C; recombinant enzyme, at pH 7.4 and 25C
0.03
5alpha-androstane-3,17-dione
-
isoform AKR1C4, at pH 7.4 and 25C
0.087 - 0.35
5alpha-androstane-3alpha,17beta-diol
0.003
5alpha-dihydroprogesterone
-
in 100 mM potassium phosphate, pH 7.0, at 25C
0.033 - 0.1153
5alpha-dihydrotestosterone
0.048 - 0.18
5beta-Androstan-3alpha-ol-17-one
0.047 - 0.3
5beta-androstane-3alpha,17beta-diol
0.002 - 0.003
5beta-pregnane-3alpha,20alpha-diol
0.083 - 0.42
9alpha,11beta-prostaglandin F2
0.000017
allopregnanolone
-
in 100 mM potassium phosphate, pH 7.0, at 25C
0.023 - 0.28
androsterone
0.032 - 0.13
chenodeoxycholic acid
0.03 - 0.2
cholic acid
0.012 - 0.085
deoxycholic acid
0.025
dihydrotestosterone
-
pH 7.5, 37C
0.032 - 0.043
lithocholic acid
0.2 - 0.58
NAD+
0.04 - 0.25
NADP+
0.0035 - 0.0392
pregn-4-en-3,20-dione
additional information
additional information
-
Kcat-value for the 17beta-oxidation of testosterone is 0.000118 1/sec, and Kcat-value for the 17beta-reduction of androstenedione is 0.00112 1/sec
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1 - 1.2
(R)-tetralol
0.68 - 1
(S)-indan-1-ol
1.2 - 1.83
(S)-tetralol
3.33
3alpha-androstanediol
-
isoform AKR1C4, at pH 7.4 and 25C
20.72
5alpha-androstane-3,17-dione
-
isoform AKR1C4, at pH 7.4 and 25C
10.3 - 135
5alpha-androstane-3alpha,17beta-diol
1.623 - 28.07
5alpha-dihydrotestosterone
53.3 - 101.7
5beta-Androstan-3alpha-ol-17-one
4.6 - 105
androsterone
28.3 - 46.7
chenodeoxycholic acid
1 - 3.3
cholic acid
8.8 - 14.2
deoxycholic acid
22.7
dihydrotestosterone
-
pH 7.5, 37C
21.7 - 31.7
lithocholic acid
166.7 - 583.3
NAD+
23.3 - 500
NADP+
4.167 - 18.4
pregn-4-en-3,20-dione
additional information
additional information
-
Kcat/Km-value for the 17beta-oxidation of testosterone is 0.183 1/sec*mM, and Kcat/Km-value for the 17beta-reduction of androstenedione is 0.817 1/sec*mM
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0028
(3,6-dihydropyridin-1(2H)-yl)(1H-indol-2-yl)methanone
Homo sapiens
-
pH 6.0, 22C
0.000037
(5-methyl-1H-indol-2-yl)(4-propylpiperidin-1-yl)methanone
Homo sapiens
-
pH 6.0, 22C
0.0029
3-pentyl-2-[[(pyridin-2-yl)methyl]sulfanyl]-7-(pyrrolidine-1-carbonyl)quinazolin-4(3H)-one
Homo sapiens
-
pH 6.0, 22C
0.0026
4-chloro-N-[(4-chlorophenyl)methyl]-5-nitro-1H-pyrazole-3-carboxamide
Homo sapiens
-
pH 6.0, 22C
0.00049
4-nitro-2-([4-[3-(trifluoromethyl)phenyl]piperazin-1-yl]methyl)phenol
Homo sapiens
-
pH 6.0, 22C
0.00029
5-(benzenesulfonyl)-2-nitrophenol
Homo sapiens
-
pH 6.0, 22C
0.00013 - 0.00015
chenodeoxycholic acid
0.0007 - 0.0009
Flufenamic acid
0.0028 - 0.0035
Hexestrol
0.0069 - 0.01
Ibuprofen
0.07 - 0.075
indomethacin
0.00007 - 0.00008
lithocholic acid
0.0016 - 0.0017
Medroxyprogesterone acetate
0.012 - 0.018
Phenolphthalein
0.00006 - 0.00008
Ursodeoxycholic acid
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
-
assay at
7.4
assay at
8
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
-
parallel expression of Siah2 and AKR1C3 in human prostate cancer tissues
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36000
-
x * 36000, SDS-PAGE
36790
-
calculated from predicted DD2 amino acid sequence of 322 residues
36850
-
calculated from predicted DD4 amino acid sequence of 321 residues
37000
-
purified DD2
39000
-
purified DD4
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 36000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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
crystal structure determination and analysis of human 3alpha-HSD3-NADP+-5alpha-androstane-3,17-dione/epiandrosterone complex, which was obtained by co-crystallization with 5alpha-DHT in the presence of NADP+. Although 5alpha-DHT is introduced during the crystallization, oxidoreduction of 5alpha-DHT occurs. The locations of 5alpha-androstane-3,17-dione and epiandrosterone are identified in the steroid-binding sites of two 3alpha-HSD3 molecules per crystal asymmetric unit. 5alpha-androstane-3,17-dione and epiandrosterone are oriented upside-down and flipped relative to each other, providing structural clues for 5alpha-DHT reverse binding in the enzyme with the generation of different products. determination and analysis of human 3alpha-HSD3-NADP+-4-androstene-3,17-dione complex crystal structure
-
crystal structure of the human 3alpha-HSD type 3 in the ternary complex with testosterone and NADP+ determined by molecular replacement method using the program EPMR, at 1.25-A resolution, binding site analysis
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purified recombinant enzyme in ternary complex with NADP+ and one inhibitor, from inhibitors 1-6, 15 mg/ml protein in 10 mM potassium phosphate pH 7.4, 500 mM NaCl, 1 mM ethylenediaminetetraacetic acid, 1 mM dithiothreitol is mixed with crystallization solution containing 0.1 M sodium citrate, pH 5.5, 0.4 M ammonium acetate, 2.5% v/v 2-methyl-2,4-pentanediol, 22-30% w/v PEG 4000 for inhibitors 1-4, and containing 0.1 M HEPES pH 6.5, 0.2 M ammonium dihydrogen phosphate, 20-25% w/v PEG 3350 for inhibitors 5 and 6, X-ray diffraction structure determination analysis at 1.55-2.81 A resolution, modelling
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Q-Sepharose column chromatography and Sephadex G-100 gel filtration
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recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, gel fitration, and ultrafiltration
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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
recombinant His-tagged AKR1C3 from Escherichia coli strain BL21
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in COS-1 cells, PC-3 cells, and LNCaP cells
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expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli cells
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expressed in Escherichia coli JM109 cells
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expressed in Huh-7 and Hep-G2 cells
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overexpression in Escherichia coli; overexpression in Escherichia coli
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overexpression of GST-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS
recombinant expression in mouse PCa cells and of FLAG-AKR1C3 in mouse Rv1 cells, recombinant expression of His-tagged AKR1C3 in Escherichia coli strain BL21
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recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
liver X receptor alpha binds specifically to the liver X receptor alpha response element and mediates transcriptional activation of the AKR1C4 gene. Liver X receptor ligand T1317 induces isoform AKR1C4 expression approximately 2.3fold
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R1881 treatment decreases AKR1C3 transcripts by about 1.5fold in Rv1 cells, AKR1C3 transcripts are androgen-repressed
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H216F
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the mutation decreases 3fold the Km for NADP+. The kinetic constants for bile acids with a 12alpha-hydroxy group are decreased 1.5-7fold and those for the other substrates are increased 1.3-9fold. The mutation decreases the stimulatory effects of the enzyme activity by sulfobromophthalein, clofibric acid and thyroxine, which increases the Km for the coenzyme and substrate of the mutant enzymes more highly than those of the wild type enzyme
H216L
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inactive
H216W
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inactive
H216Y
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the mutation decreases 3fold the Km for NADP+. The kinetic constants for bile acids with a 12alpha-hydroxy group are decreased 1.5-7fold and those for the other substrates are increased 1.3-9fold. The mutation decreases the stimulatory effects of the enzyme activity by sulfobromophthalein, clofibric acid and thyroxine, which increases the Km for the coenzyme and substrate of the mutant enzymes more highly than those of the wild type enzyme
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
additional information
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when a specific siRNA is used to suppress 3alpha-HSD3 expression without interfering with 3alpha-HSD4, which shares a highly homologous active site, the 5alpha-DHT concentration increases, whereas MCF-7 cell growth is suppressed. Downregulation of 3alpha-HSD3 decreases MCF-7 breast cancer cell growth
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
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testosterone is converted to 5alpha-dihydrotestosterone, which is present at high concentrations in patients with castration resistant prostate cancer (CRPC). Inhibition of 17beta-HSD5 is therefore considered to be a promising therapy for treating CRPC. High-throughput inhibitor screening, overview