Information on EC 1.1.1.145 - 3beta-hydroxy-DELTA5-steroid dehydrogenase and Organism(s) Homo sapiens

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The taxonomic range for the selected organisms is: Homo sapiens

The enzyme appears in selected viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.1.1.145
-
RECOMMENDED NAME
GeneOntology No.
3beta-hydroxy-DELTA5-steroid dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a 3beta-hydroxy-DELTA5-steroid + NAD+ = a 3-oxo-DELTA5-steroid + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
androgen biosynthesis
-
-
androstenedione degradation
-
-
cholesterol degradation to androstenedione II (cholesterol dehydrogenase)
-
-
progesterone biosynthesis
-
-
sitosterol degradation to androstenedione
-
-
androgen and estrogen metabolism
-
-
Steroid hormone biosynthesis
-
-
Steroid degradation
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
3beta-hydroxy-DELTA5-steroid:NAD+ 3-oxidoreductase
This activity is found in several bifunctional enzymes that catalyse the oxidative conversion of Delta5-3-hydroxy steroids to a Delta4-3-oxo configuration. This conversion is carried out in two separate, sequential reactions; in the first reaction, which requires NAD+, the enzyme catalyses the dehydrogenation of the 3beta-hydroxy steroid to a 3-oxo intermediate. In the second reaction the reduced coenzyme, which remains attached to the enzyme, activates the isomerization of the Delta5 form to a Delta4 form (cf. EC 5.3.3.1, steroid Delta-isomerase). Substrates include dehydroepiandrosterone (which is converted into androst-5-ene-3,17-dione), pregnenolone (converted to progesterone) and cholest-5-en-3-one, an intermediate of cholesterol degradation.
CAS REGISTRY NUMBER
COMMENTARY hide
9044-85-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-1-indanol + NADP+
? + NADPH
show the reaction diagram
-
-
-
-
r
(S)-1-tetralol + NADP+
? + NADPH
show the reaction diagram
-
-
-
-
r
(S)-phenyl-1-butanol + NADP+
? + NADPH
show the reaction diagram
-
-
-
-
r
1,4-naphthoquinone + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
1-phenylisatin + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
16alpha-hydroxy-dehydroepiandrosterone + NAD+
?
show the reaction diagram
-
-
-
-
?
16alpha-hydroxy-dehydroepiandrosterone + NAD+
? + NADH
show the reaction diagram
-
-
-
-
?
16beta-hydroxy-dehydroepiandrosterone + NAD+
?
show the reaction diagram
-
-
-
-
?
16beta-hydroxy-dehydroepiandrosterone + NAD+
? + NADH
show the reaction diagram
-
-
-
-
?
17alpha-hydroxypregnenolone + NAD+
17alpha-hydroxy-pregn-5-en-3,20-dione + NADH
show the reaction diagram
2,3-heptanedione + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
2,3-hexanedione + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
2,3-pentanedione + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
r
3beta,17alpha-dihydroxy-5-pregnen-20-one + NAD(P)+
17alpha-hydroxypregn-5-ene-3,20-dione + NAD(P)H
show the reaction diagram
-
-
-
-
?
4,4'-dimethylbenzil + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
4-benzoylpyridine + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
4-hexanoylpyridine + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
4-nitrobenzaldehyde + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
5-pregnen-3beta-ol-2-one + NAD+
progesterone + NADH
show the reaction diagram
-
-
-
-
?
5alpha-androstan-3beta-ol-17-one + NADP+
5alpha-androstan-3,17-dione + NADPH
show the reaction diagram
5alpha-androstane-3,17-dione + NADPH
5alpha-androstane-3beta-ol-17-one + NADP+
show the reaction diagram
-
-
-
-
r
5alpha-androstane-3beta,17beta-diol + NADP+
5alpha-androstane-17beta-ol-3-one + NADPH
show the reaction diagram
-
-
-
-
r
5alpha-dihydrotestosterone + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
5alpha-pregnane-3,20-dione + NADPH
5beta-pregnane-3,20-dione + NADPH
show the reaction diagram
-
-
-
-
r
5alpha-pregnane-3beta-ol-20-one + NADP+
5alpha-pregnane-3,20-dione + NADPH
show the reaction diagram
-
-
-
-
r
5beta-androstan-17beta-ol-3-one + NADPH
5beta-androstane-3beta,17beta-diol + NADP+
show the reaction diagram
-
-
-
-
r
5beta-androstan-3beta-ol-17-one + NADP+
5beta-androstane-3,17-dione + NADPH
show the reaction diagram
-
-
-
-
r
5beta-androstane-3,17-dione + NADPH
5beta-androstan-3beta-ol-17-one + NADP+
show the reaction diagram
-
-
-
-
r
5beta-androstane-3beta,17beta-diol + NADP+
5beta-androstan-17beta-ol-3-one + NADPH
show the reaction diagram
-
-
-
-
r
5beta-pregnan-20alpha-ol-3-one + NADPH + H+
5beta-pregnan-3beta,20alpha-diol + NADP+
show the reaction diagram
-
-
-
-
r
5beta-pregnan-3beta-ol-20-one + NADP+
5beta-pregnan-3,20-dione + NADPH
show the reaction diagram
-
-
-
-
r
5beta-pregnane-3,20-dione + NADPH + H+
5beta-pregnan-3beta-ol-20-one + NADP+
show the reaction diagram
-
-
-
-
r
5beta-pregnane-3beta,20alpha-diol + NADP+
5beta-pregnane-20alpha-ol-3-one + NADPH + H+
show the reaction diagram
-
-
-
-
r
6-tert-butyl-2,3-epoxy-5-cyclohexene-1,4-dione + NADPH
? + NADP+
show the reaction diagram
-
major product has S-configuration
-
-
r
9,10-phenanthrenequinone + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
all-trans-retinal + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
all-trans-retinol + NADP+
? + NADPH
show the reaction diagram
-
-
-
-
r
androst-5-en-3,17-dione + NADH
dehydroepiandrosterone + NAD+
show the reaction diagram
-
-
-
-
r
benzil + NADPH
(R)-benzoin + NADP+
show the reaction diagram
-
enantiomeric purity of 97% enantiomeric excess
-
-
r
dehydroepiandrosterone + NAD+
?
show the reaction diagram
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH
show the reaction diagram
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
show the reaction diagram
-
-
-
-
?
dehydrolithocholic acid + NADPH
isolithocholic acid + NADP+
show the reaction diagram
-
-
-
-
r
DELTA5-3beta-hydroxysteroid + NAD+
DELTA4-3-oxosteroid + NADH + H+
show the reaction diagram
DELTA5-androstenediol + NAD+
testosterone + NADH + H+
show the reaction diagram
-
-
-
-
?
diacetyl + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
dimethyl-2-oxo-glutarate + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
epiandrosterone + NAD+
5alpha-androstan-3,17-dione + NADH + H+
show the reaction diagram
-
-
-
-
r
hexanophenone + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
isatin + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
isolithocholic acid + NADP+
dehydrolithocholic acid + NADPH
show the reaction diagram
-
-
-
-
r
menadione + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
n-butyrophenone + NADPH
(S)-1-phenyl1-butanol + NADP+
show the reaction diagram
-
major product
-
-
r
pregnenolone + NAD(P)+
pregn-5-en-3,20-dione + NAD(P)H
show the reaction diagram
-
-
-
-
?
pregnenolone + NAD+
pregn-5-en-3,20-dione + NADH
show the reaction diagram
-
-
-
?
pregnenolone + NAD+
pregn-5-en-3,20-dione + NADH + H+
show the reaction diagram
-
-
-
-
?
pregnenolone + NAD+
progesterone + NADH
show the reaction diagram
pregnenolone + NAD+
progesterone + NADH + H+
show the reaction diagram
pyridine-4-aldehyde + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
testosterone + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
valerophenone + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
r
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
17alpha-hydroxypregnenolone + NAD+
17alpha-hydroxy-pregn-5-en-3,20-dione + NADH
show the reaction diagram
5-pregnen-3beta-ol-2-one + NAD+
progesterone + NADH
show the reaction diagram
-
-
-
-
?
dehydroepiandrosterone + NAD+
?
show the reaction diagram
P14060, P26439
isozyme type 1
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH
show the reaction diagram
DELTA5-3beta-hydroxysteroid + NAD+
DELTA4-3-oxosteroid + NADH + H+
show the reaction diagram
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, molecular mechanisms of enzyme regulation involving the Nur77 family of immediate-early orphan nuclear receptors, e.g. Nur77 targets the promoter of gene HSD3B2 in testicular Leygig cells, overview
-
-
?
DELTA5-androstenediol + NAD+
testosterone + NADH + H+
show the reaction diagram
-
-
-
-
?
epiandrosterone + NAD+
5alpha-androstan-3,17-dione + NADH + H+
show the reaction diagram
-
-
-
-
r
pregnenolone + NAD(P)+
pregn-5-en-3,20-dione + NAD(P)H
show the reaction diagram
-
-
-
-
?
pregnenolone + NAD+
pregn-5-en-3,20-dione + NADH
show the reaction diagram
P14060, P26439
-
-
-
?
pregnenolone + NAD+
pregn-5-en-3,20-dione + NADH + H+
show the reaction diagram
-
-
-
-
?
pregnenolone + NAD+
progesterone + NADH + H+
show the reaction diagram
P14060, P26439
isozyme type 2
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
NADPH
-
-
additional information
-
no significant activity with NAD+/NADH
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
no effect of Ca2+, Sr2+, Ba2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(+)-gossypol
-
potent inhibitor
(-)-gossypol
-
potent inhibitor
17beta-acetoxy-trilostane
-
-
4alpha,5alpha-epoxy-testosterone
-
-
5,10-Secoestr-4-yne-3,10,17-trione
-
irreversible
6-hydroxyflavone
-
0.00056 mM, 50% inhibition; competitive
abiraterone
-
-
apigenin
-
0.13 mM, 50% inhibition
Biochanin A
Cd2+
-
1 mM: strong
Cetylpyridinium chloride
-
-
Co2+
-
1 mM: weak, 100 mM: strong
Cu2+
-
1 mM: strong
cyanoketone
-
0.0000076 mM, 50% inhibition; competitive
daidzein
-
0.0033 mM, 50% inhibition
drospirenone
-
-
epostane
Fe2+
-
1 mM: weak, 100 mM: strong
flavanones
-
diverse derivatives, inhibitory potential determination, overview
flavones
-
diverse derivatives, inhibitory potential determination, overview
formononetin
-
0.0032 mM, 50% inhibition
genistein
isoflavones
-
diverse derivatives, inhibitory potential determination, overview
lauric acid
-
0.05 mM, less than 10% inhibition
Mn2+
-
1 mM: weak
myricetin
-
0.011 mM, 50% inhibition
myristic acid
-
0.05 mM, less than 10% inhibition
nestorone
-
-
Ni2+
-
1 mM: weak, 100 mM: strong
nomegestrol acetate
-
-
phenylhexane
-
-
quercetin
trilostane
Zn2+
-
1 mM: strong
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.24
(S)-1-indanol
-
pH 7.4, 25C
0.093
(S)-1-tetralol
-
pH 7.4, 25C
0.0084
(S)-phenyl-1-butanol
-
pH 7.4, 25C
0.089
1,4-Naphthoquinone
-
pH 7.4, 25C
0.0052
1-phenylisatin
-
pH 7.4, 25C
0.004 - 0.0196
16alpha-hydroxy-dehydroepiandrosterone
0.0014 - 0.0184
16beta-hydroxy-dehydroepiandrosterone
0.0035 - 0.018
17alpha-hydroxypregnenolone
0.2
2,3-heptanedione
-
pH 7.4, 25C
0.55
2,3-Hexanedione
-
pH 7.4, 25C
10
2,3-Pentanedione
-
pH 7.4, 25C
0.0015
4,4'-dimethylbenzil
-
pH 7.4, 25C
1
4-benzoylpyridine
-
pH 7.4, 25C
0.16
4-hexanoylpyridine
-
pH 7.4, 25C
13
4-nitrobenzaldehyde
-
pH 7.4, 25C
0.0144 - 0.0147
5-androstene-3,17-dione
0.0167 - 0.0179
5alpha-Androstan-3beta-ol-17-one
0.028
5alpha-androstane-3,17-dione
-
pH 7.4, 25C
0.016
5alpha-dihydrotestosterone
-
pH 7.4, 25C
0.0049
5beta-androstan-17beta-ol-3-one
-
pH 7.4, 25C
0.0079
5beta-androstan-3beta-ol-17-one
-
pH 7.4, 25C
0.01
5beta-androstane-3,17-dione
-
pH 7.4, 25C
0.0041
5beta-androstane-3beta,17beta-diol
-
pH 7.4, 25C
0.0097
5beta-pregnan-20alpha-ol-3-one
-
pH 7.4, 25C
0.008
5beta-Pregnan-3beta-ol-20-one
-
pH 7.4, 25C
0.0029
5beta-pregnane-3,20-dione
-
pH 7.4, 25C
0.008
5beta-pregnane-3beta,20alpha-diol
-
pH 7.4, 25C
0.12
6-tert-butyl-2,3-epoxy-5-cyclohexene-1,4-dione
-
pH 7.4, 25C
0.0036
9,10-phenanthrenequinone
-
pH 7.4, 25C
0.024
all-trans-retinal
-
pH 7.4, 25C
0.005
benzil
-
pH 7.4, 25C
0.0007 - 0.0643
dehydroepiandrosterone
0.035
Dehydrolithocholic acid
-
pH 7.4, 25C
54
diacetyl
-
pH 7.4, 25C
1.4
dimethyl-2-oxo-glutarate
-
pH 7.4, 25C
0.036
hexanophenone
-
pH 7.4, 25C
0.32
isatin
-
pH 7.4, 25C
0.18
menadione
-
pH 7.4, 25C
0.048
n-butyrophenone
-
pH 7.4, 25C
0.0034 - 0.0863
NAD+
0.0025 - 0.0126
NADH
0.0003
NADP+
-
pH 7.4, 25C
0.0003
NADPH
-
pH 7.4, 25C
0.00007 - 0.05
pregnenolone
21
Pyridine-4-aldehyde
-
pH 7.4, 25C
0.054
valerophenone
-
pH 7.4, 25C
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.18
(S)-1-indanol
-
pH 7.4, 25C
0.25
(S)-1-tetralol
-
pH 7.4, 25C
0.075
(S)-phenyl-1-butanol
-
pH 7.4, 25C
0.88
1,4-Naphthoquinone
-
pH 7.4, 25C
41.7
1-phenylisatin
-
pH 7.4, 25C
0.07 - 0.15
16alpha-hydroxy-dehydroepiandrosterone
0.04 - 0.15
16beta-hydroxy-dehydroepiandrosterone
40.2
2,3-heptanedione
-
pH 7.4, 25C
16.3
2,3-Hexanedione
-
pH 7.4, 25C
26.7
2,3-Pentanedione
-
pH 7.4, 25C
3.7
4,4'-dimethylbenzil
-
pH 7.4, 25C
2
4-benzoylpyridine
-
pH 7.4, 25C
33.3
4-hexanoylpyridine
-
pH 7.4, 25C
16.7
4-nitrobenzaldehyde
-
pH 7.4, 25C
0.097
5alpha-androstane-3,17-dione
-
pH 7.4, 25C
0.095
5alpha-dihydrotestosterone
-
pH 7.4, 25C
0.083
5beta-androstan-17beta-ol-3-one
-
pH 7.4, 25C
0.017
5beta-androstan-3beta-ol-17-one
-
pH 7.4, 25C
0.077
5beta-androstane-3,17-dione
-
pH 7.4, 25C
0.017
5beta-androstane-3beta,17beta-diol
-
pH 7.4, 25C
0.07
5beta-pregnan-20alpha-ol-3-one
-
pH 7.4, 25C
0.041
5beta-Pregnan-3beta-ol-20-one
-
pH 7.4, 25C
0.063
5beta-pregnane-3,20-dione
-
pH 7.4, 25C
0.014
5beta-pregnane-3beta,20alpha-diol
-
pH 7.4, 25C
2
6-tert-butyl-2,3-epoxy-5-cyclohexene-1,4-dione
-
pH 7.4, 25C
11.8
9,10-phenanthrenequinone
-
pH 7.4, 25C
0.05
all-trans-retinal
-
pH 7.4, 25C
31.7
benzil
-
pH 7.4, 25C
0.02 - 0.217
dehydroepiandrosterone
0.043
Dehydrolithocholic acid
-
pH 7.4, 25C
28.3
diacetyl
-
pH 7.4, 25C
33.3
dimethyl-2-oxo-glutarate
-
pH 7.4, 25C
2.7
hexanophenone
-
pH 7.4, 25C
31.7
isatin
-
pH 7.4, 25C
0.65
menadione
-
pH 7.4, 25C
0.85
n-butyrophenone
-
pH 7.4, 25C
0.02 - 0.187
NAD+
0.31 - 1.65
NADH
4.7
Pyridine-4-aldehyde
-
pH 7.4, 25C
2
valerophenone
-
pH 7.4, 25C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.83 - 19.1
16alpha-hydroxy-dehydroepiandrosterone
7.66 - 35.5
16beta-hydroxy-dehydroepiandrosterone
0.0028 - 14.83
dehydroepiandrosterone
0.83 - 4.83
NAD+
106 - 280
NADH
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00043
17beta-acetoxy-trilostane
-
in 0.02 M potassium phosphate buffer, pH 7.4
0.0028 - 0.00434
4alpha,5alpha-epoxy-testosterone
0.00019
6-hydroxyflavone
-
37C, pH 7.4; recombinant enzyme
0.00000067
cyanoketone
-
37C, pH 7.4; recombinant enzyme
0.000232
drospirenone
-
pH and temperature not specified in the publication
0.000007 - 0.00184
epostane
0.00012
genistein
-
37C, pH 7.4; recombinant enzyme
0.0000095
nestorone
-
pH and temperature not specified in the publication
0.000029
nomegestrol acetate
-
pH and temperature not specified in the publication
0.00001 - 0.016
trilostane
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00445
(+)-gossypol
Homo sapiens;
-
in PBS buffer, pH 7.2
0.00277
(-)-gossypol
Homo sapiens;
-
in PBS buffer, pH 7.2
0.0066
Cetylpyridinium chloride
Homo sapiens;
-
pH 7.4, 25C
0.0002
epostane
Homo sapiens;
-
in 0.02 M potassium phosphate buffer, pH 7.4
0.0059
genistein
Homo sapiens;
-
pH 7.4, 25C
0.023
phenylhexane
Homo sapiens;
-
pH 7.4, 25C
0.0027
quercetin
Homo sapiens;
-
pH 7.4, 25C
0.0049
trilostane
Homo sapiens;
-
in 0.02 M potassium phosphate buffer, pH 7.4
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0062
-
wild type, microsomes
0.0091
-
placental microsomes, 3beta-HSD activity
0.0099
-
wild type, mitochondria
0.0145
-
Sf9-cells, 3beta-HSD activity
0.0172
-
placental microsomes, isomerase activity
0.0225
-
283-310 deletion, cytosol
0.0257
-
Sf9-cells, isomerase activity
0.0524
-
-
0.155
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
peak 3betaHSD2 activity at pH 4.5 with 2fold more progesterone synthesized at pH 4.5 than at pH 3.5 and 7.4
5.5
-
dimethyl-2-oxoglutarate reductase activity
6.5 - 10
-
broad, S-1-tetralol dehydrogenase activity
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
52% of maximal activity
7.4 - 9.7
additional information
-
metabolic conversion assays indicate peak 3betaHSD2 activity at pH 4.5 with 2fold more progesterone synthesized at pH 4.5 than at pH 3.5 and 7.4. Increasing the 3betaSD2 concentration from 1 to 40 microg results in a 7fold increase in progesterone at pH 4.5, but no change at pH 7.4. At pH 4-5, 3betaHSD2 takes on a molten globule conformation that promotes the dual functionality of the enzyme, i.e. 3 beta-hydroxysteroid dehydrogenase and Delta 5->4-isomerase activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
40 - 45
-
-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 55
-
-
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
abdominal, expression level determination, especially in female aorta with mild atherosclerotic changes
Manually annotated by BRENDA team
-
primary culture
Manually annotated by BRENDA team
-
adrenal cell line
Manually annotated by BRENDA team
-
induction by peroxisome-proliferator-activated receptor alpha ligands
Manually annotated by BRENDA team
-
3beta-HSD II
Manually annotated by BRENDA team
isozyme 3beta-HSD1
Manually annotated by BRENDA team
-
3beta-HSD I
Manually annotated by BRENDA team
-
in amnion epithelium immunoreactive 3betaHSD is found from first to third trimester and term
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
in amnion epithelium, chorion trophoblast and invasive trophoblast immunoreactive 3betaHSD is found from first to third trimester and term
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
three-dimensional structure modeling of enzyme in ternary complex with NAD+ and androstenedione using crystals structures of enzymes from other organisms, substrate binding and active site residue determination, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
modified enzyme
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 7.4
-
circular dichroism shows that between pH 7.4 and 4.5, 3betaHSD2 retains its primarily alpha-helical character with a decrease in beta-helical content at lower pH values, whereas the beta-sheet content remains unchanged throughout. Titrating the pH back to 7.4 restores the original conformation; incubation with guanidinum hydrochloride shows a three-step cooperative unfolding of 3betaHSD2 from pH 7.4 to 4.5. With further decreases in pH, increasing concentrations of GdmHCl lead to rapid two-step unfolding that may represent complete loss of structure. Between pH 4 and 5, the two intermediate states appear stable. Stopped-flow kinetics show slower unfolding at around pH 4, where the protein is in a pseudostable state. At pH 4-5, 3betaHSD2 takes on a molten globule conformation that promotes the dual functionality of the enzyme
725511
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42
-
melting temperature in the absence of lipid; Tm in the absence of any lipid
65
-
complete denaturation
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified from Sf9 cells in the presence of the low-CMC detergent, Igepal CO 720, using a DEAE ion exchange column
-
recombinant 3beta-HSD1
-
recombinant 3beta-HSD; recombinant 3beta-HSD type 2
-
recombinant soluble wild-type and mutant isozyme 3beta-HSD1 from Sf9 insect cells; recombinant soluble wild-type and mutant isozyme 3beta-HSD2 from Sf9 insect cells
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in MA-10 cells; gene HSD3B2, analysis of expression and its regulation involving Nur77 factors in H295R and MA-10 cell lines, overview
-
expressed in Sf9 cells using the baculovirus system
-
expressed in Sf9 cells using the baculovirus system; expressed in Sf9 cells using the baculovirus system
-
expressed in Sf9 cells using the baculovirus system; expression in baculoviral system
-
expressed in Sf9 insect cells
-
expression analysis in abdominal aorta of 28 samples from japanese patients
-
expression in Sf9 cells
-
expression in Sf9 cells; expression in Sf9 insect cells
-
expression in Sf9 cells; expression of soluble wild-type and mutant isozyme 3beta-HSD1 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system; expression of soluble wild-type and mutant isozyme 3beta-HSD2 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system
expression in Sf9 cells; expression of the isozyme type II in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system
-
expression in Sf9 insect cells
-
expression in Spodoptera fungiperda insect cells
-
expression of soluble wild-type and mutant isozyme 3beta-HSD1 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system; expression of soluble wild-type and mutant isozyme 3beta-HSD2 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system
overexpression with his6-tag, using a baculovirus expression system
-
transient expression of HSD I and II cDNA in HeLa cells
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
isoform HSD3B1 is expressed specifically within the adrenal zona glomerulosa. Angiotensin II triggers expression of HSD3B1but not that of isoform HSD3B2 in adrenocortical H295R cells. The induction of HSD3B1 involves de novo protein synthesis of the nuclear orphan receptors NGFIB and NURR1
-
the mRNA level of the enzyme is higher in tumor tissue compared with those of the normal tissue
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C111A
-
contrary to wild-type, presence of 2-mercaptoethanol does not decrease Km value for substrate and NAD+
C111S
-
contrary to wild-type, presence of 2-mercaptoethanol does not decrease Km value for substrate and NAD+
C72F
-
contrary to wild-type, presence of 2-mercaptoethanol does not decrease Km value for substrate and NAD+
C72S
-
contrary to wild-type, presence of 2-mercaptoethanol does not decrease Km value for substrate and NAD+
C83A
-
2-5fold increase in Km values
C83S
-
no catalytic acitivity; similar to wild-type, presence of 2-mercaptoethanol results in 2.5fold decrease of Km values
D241N
-
39% of wild-type activity
D257L
-
trace activity
D258L
-
trace activity
D265N
-
12% of wild-type activity
D35A/K36R
-
the D35A/K36R mutant enzyme of 3beta-HSD1 has no measurable 3beta-HSD activity with NADH as the cofactor and now uses NADPH as the cofactor and NADH specificity of the wild-type 1 isomerase is completely lost
D36A/K37R
-
uses NADP+ as cofactor, no activity with NAD+
D61N
-
D61N mutation produces a 1.3fold increase in Km for the dehydroepiandrosterone substrate, a 1.9fold increase in Km for NAD+ utilization by the 3beta-HSD activity and little change in isomerase kinetics for either substrate or cofactor
D61V
-
mutant shows no enzymatic activity
DELTA283-310
-
cytosolic enzyme
E126L
site-directed mutagenesis, functional characterization
E192A
-
the E192A mutation causes an 8.2fold increase in Km (dehydroepiandrosterone) and no change in the Km for NAD+
H154F
no activity
H232A
site-directed mutagenesis, functional characterization
L341P
-
mutation in HSD3B2 gene identified in neonate with classical 3beta-hydroxysteroid dehydrogenase type II deficiency and under-virilization. About 6% of wild-type activity, putative structural alteration in protein
M187T
-
the 3beta-HSD1 mutant shows 16fold lower affinity inhibition by trilostane compared to the wild type enzyme, and produces a 2.2fold decrease in the Km value for the NAD+ utilization by of 3beta-HSD1 and a 9.2fold decrease in the Km measured for NADH
N100A
site-directed mutagenesis, functional characterization
N100S
site-directed mutagenesis, functional characterization
N323L
site-directed mutagenesis, functional characterization
Q105M
site-directed mutagenesis of isozyme 1, the mutants show altered kinetics compared to the wild-type isozymes; site-directed mutagenesis of isozyme 2, the mutants show altered kinetics compared to the wild-type isozymes
Q105M1
5fold decrease in catalytic efficiency; mutant retains the lower affinity profile of inhibition by epostane that is observed with wild-type 3beta-HSD2
Q240R
site-directed mutagenesis of isozyme 1, the mutant shows altered kinetics compared to the wild-type isozyme
R195R
-
the P195R mutation shifts the low-affinity, noncompetitive inhibition profile of 3beta-HSD2 to a high affinity (Ki= 0.19 microM), competitive inhibition profile. The P195R mutant has a 12.2fold lower Km value for 16beta-hydroxy-dehydroepiandrosterone compared to wild-type 3beta-HSD2. The coenzyme values measured for P195R mutant are similar to those measured for wildtype 3beta-HSD2
R240Q
site-directed mutagenesis of isozyme 2, the mutant shows altered kinetics compared to the wild-type isozyme
R335stop
-
mutation in HSD3B2 gene identified in neonate with classical 3beta-hydroxysteroid dehydrogenase type II deficiency and under-virilization. No conversion of pregnenolone or dehydroepiandrosterone
S123A
-
24% of wild-type 3beta-HSD1 activity
S124A
-
no detectable 3beta-HSD1 activity
S124T
-
mutant shows activity similar to the wild type enzyme
S322A
site-directed mutagenesis, functional characterization
T177N
-
significantly improved stability at cold temperature, presence of NAD+ is required for complete stability at 0C
T8A
-
mutant shows almost similar kinetic values as wild-type
W355stop
-
mutation in HSD3B2 gene identified in neonate with classical 3beta-hydroxysteroid dehydrogenase type II deficiency and under-virilization. No conversion of pregnenolone or dehydroepiandrosterone
additional information
-
identification of 17 single nucleotide polymorphisms in isoforms Hsd3B1 and Hsd3B2. Analysis of polymorphism locations, alterations in nucleotide and amino acid sequences and frequencies of the polymorphisms. None of the polymorphisms alters cellular localization of the enzyme
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
between pH 7.4 and 4.5, isoform 3betaHSD2 retains its primarily-helical character with a decrease in alpha-helical content at lower pH values, whereas the beta-sheet content remains unchanged throughout. Titrating the pH back to 7.4 restores the original conformation within 25 min. Metabolic conversion assays indicate peak 3betaHSD2 activity at pH 4.5 with 2fold more progesterone synthesized at pH 4.5 than at pH 3.5 and 7.4. Increasing the 3betaSD2 concentration from 1 to 40 microg results in a 7fold increase in progesterone at pH 4.5, but no change at pH 7.4. Incubation with guanidinum hydrochloride shows a three-step cooperative unfolding of 3betaHSD2 from pH 7.4 to 4.5, possibly due to the native state unfolding to the intermediate ion core state. With further decreases in pH, increasing concentrations of guanidinum hydrochloride lead to rapid two-step unfolding that may represent complete loss of structure
-
enzyme interacts strongly with a mixture of dipalmitoylphosphatidylglycerol and dipalmitoylphosphatidylcholine. It becomes less stable when incubated with the individual lipids. Dipalmitoylphosphatidylglycerol, alone or in combination with dipalmitoylphosphatidylcholine, leads to a decrease in alpha-helical content without an effect on the beta-sheet conformation. With the exception of the 20 N-terminal amino acids, mixed vesicles protect the enzyme from trypsin digestion. Protein incubated with dipalmitoylphosphatidylcholine is only partially protected
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine