Any feedback?
Information on EC 1.1.1.145 - 3beta-hydroxy-DELTA5-steroid dehydrogenase and Organism(s) Homo sapiens and UniProt Accession P14060
for references in articles please use BRENDA:EC1.1.1.145Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
1.1.1.145 3beta-hydroxy-DELTA5-steroid dehydrogenaseIUBMB Comments
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.
Specify your search results
Word Map
- 1.1.1.145
-
steroidogenic
- adrenal
-
steroidogenesis
- star
- cyp11a1
- progesterone
- androgen
- ovary
- ovarian
- gonad
- follicle
-
granulosa
- leydig
- cortisol
-
adrenocortical
- dehydroepiandrosterone
- aldosterone
- androstenedione
- aromatase
- dheas
- 21-hydroxylase
- hsd17b3
- pregnenolone
-
reticularis
- srd5a2
-
lhcgr
-
nci-h295r
- p450scc
- cyp21
- genitalia
- hyperandrogenism
- akr1c3
- p450arom
-
adrenarche
- hypospadias
- 17-hydroxypregnenolone
- 17-hydroxyprogesterone
-
pubarche
-
steroidogenesis-related
-
aldosterone-producing
-
salt-wasting
- 17alpha-hydroxylase
- sult2a1
-
virilization
- medicine
- diagnostics
-
cortisol-producing
- nutrition
The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
hsd3b2, hsd3b, dhrs4, 3-beta-hydroxysteroid dehydrogenase, hsd3b6, 3betahsd2, 3beta hydroxysteroid dehydrogenase, 3beta-hydroxysteroid dehydrogenase type ii, 3beta-hydroxysteroid dehydrogenase type 2, 3beta-hsd type ii, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
3-beta-hydroxy-5-ene steroid dehydrogenase
-
-
-
-
3-beta-hydroxy-delta(5)-steroid dehydrogenase
-
-
-
-
3beta-HSD1
3beta-HSD2
3beta-HSDH
-
-
-
-
3beta-hydroxy steroid dehydrogenase/5-ene-4-ene isomerase
-
-
-
-
3beta-hydroxy steroid dehydrogenase/isomerase
-
-
-
-
3beta-hydroxy-5-ene steroid dehydrogenase
-
-
-
-
3beta-hydroxy-5-ene-steroid dehydrogenase
-
-
-
-
3beta-hydroxy-5-ene-steroid oxidoreductase
-
-
-
-
3beta-hydroxy-DELTA5-C27-steroid dehydrogenase/isomerase
-
-
-
-
3beta-hydroxy-DELTA5-C27-steroid oxidoreductase
-
-
-
-
3beta-hydroxysteroid dehydrogenase
3beta-hydroxysteroid dehydrogenase type 1
3beta-hydroxysteroid dehydrogenase type 2
3betaHSD2
5-ene-3-beta-hydroxysteroid dehydrogenase
-
-
-
-
5alpha-dihydrotestosterone 3beta-hydroxysteroid dehydrogenase
-
-
-
-
dehydrogenase, 3beta-hydroxy-DELTA5-steroid
-
-
-
-
DELTA5-3beta-hydroxysteroid dehydrogenase
-
-
-
-
HSD3B1
HSD3B2
progesterone reductase
-
-
-
-
steroid-DELTA5-3beta-ol dehydrogenase
-
-
-
-
additional information
the enzyme belongs to the short chain dehydrogenase/reductase family
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a 3beta-hydroxy-DELTA5-steroid + NAD+ = a 3-oxo-DELTA5-steroid + NADH + H+
catalytic residues are Lys159 and Tyr154, structure/function analysis of isozyme 1
a 3beta-hydroxy-DELTA5-steroid + NAD+ = a 3-oxo-DELTA5-steroid + NADH + H+
substrate binding and active site residues, overview
a 3beta-hydroxy-DELTA5-steroid + NAD+ = a 3-oxo-DELTA5-steroid + NADH + H+
structure-function relationship
-
a 3beta-hydroxy-DELTA5-steroid + NAD+ = a 3-oxo-DELTA5-steroid + NADH + H+
catalytic residues are Lys159 and Tyr154, structure/function analysis of isozyme 2
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY SOURCE
PATHWAYS
KEGG
-
-, -, -, -, -, -, -, -, -
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
9044-85-3
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
dehydroepiandrosterone + NAD+
5-androstene-3,17-dione + NADH + H+
-
-
-
?
3beta,17alpha-dihydroxy-5-pregnen-20-one + NAD(P)+
17alpha-hydroxypregn-5-ene-3,20-dione + NAD(P)H
-
-
-
-
?
5alpha-androstane-3,17-dione + NADPH
5alpha-androstane-3beta-ol-17-one + NADP+
-
-
-
-
r
5alpha-androstane-3beta,17beta-diol + NADP+
5alpha-androstane-17beta-ol-3-one + NADPH
-
-
-
-
r
5alpha-pregnane-3beta-ol-20-one + NADP+
5alpha-pregnane-3,20-dione + NADPH
-
-
-
-
r
5beta-androstan-17beta-ol-3-one + NADPH
5beta-androstane-3beta,17beta-diol + NADP+
-
-
-
-
r
5beta-androstan-3beta-ol-17-one + NADP+
5beta-androstane-3,17-dione + NADPH
-
-
-
-
r
5beta-androstane-3,17-dione + NADPH
5beta-androstan-3beta-ol-17-one + NADP+
-
-
-
-
r
5beta-androstane-3beta,17beta-diol + NADP+
5beta-androstan-17beta-ol-3-one + NADPH
-
-
-
-
r
5beta-pregnan-20alpha-ol-3-one + NADPH + H+
5beta-pregnan-3beta,20alpha-diol + NADP+
-
-
-
-
r
5beta-pregnan-3beta-ol-20-one + NADP+
5beta-pregnan-3,20-dione + NADPH
-
-
-
-
r
5beta-pregnane-3,20-dione + NADPH + H+
5beta-pregnan-3beta-ol-20-one + NADP+
-
-
-
-
r
5beta-pregnane-3beta,20alpha-diol + NADP+
5beta-pregnane-20alpha-ol-3-one + NADPH + H+
-
-
-
-
r
6-tert-butyl-2,3-epoxy-5-cyclohexene-1,4-dione + NADPH
? + NADP+
-
major product has S-configuration
-
-
r
androst-5-en-3,17-dione + NADH
dehydroepiandrosterone + NAD+
-
-
-
r
benzil + NADPH
(R)-benzoin + NADP+
-
enantiomeric purity of 97% enantiomeric excess
-
-
r
dehydroepiandrosterone + NAD+
5-androstene-3,17-dione + NADH + H+
-
-
-
?
pregnenolone + NAD(P)+
pregn-5-en-3,20-dione + NAD(P)H + H+
-
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
?
17alpha-hydroxypregnenolone + NAD+
17alpha-hydroxy-pregn-5-en-3,20-dione + NADH
-
-
-
?
17alpha-hydroxypregnenolone + NAD+
17alpha-hydroxy-pregn-5-en-3,20-dione + NADH
-
-
-
-
?
5alpha-androstan-3beta-ol-17-one + NADP+
5alpha-androstan-3,17-dione + NADPH
-
-
-
-
?
5alpha-androstan-3beta-ol-17-one + NADP+
5alpha-androstan-3,17-dione + NADPH
-
-
-
-
r
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
r
DELTA5-3beta-hydroxysteroid + NAD+
DELTA4-3-oxosteroid + NADH + H+
-
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
-
-
?
DELTA5-3beta-hydroxysteroid + NAD+
DELTA4-3-oxosteroid + NADH + H+
-
the enzyme performs the 3beta-hydroxysteroid dehydrogenase and DELTA5-DELTA4 isomerase reactions
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones catalyzing the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
-
-
?
additional information
?
-
-
the enzyme is essential for biosynthesis of all classes of steroid hormones catalyzing the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
-
-
?
additional information
?
-
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
-
the enzyme is involved in steroid production and/or metabolism
-
-
?
additional information
?
-
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
-
bifunctional enzyme that also catalyzes the reaction of the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1
-
-
?
additional information
?
-
-
enzyme exhibits high stereo-selectivity yielding 3-beta-hydroxysteroids from 3-ketosteroids
-
-
?
additional information
?
-
using fluorescence resonance energy transfer, it is shown that 3betaHSD2 interacts strongly with a mixture of dipalmitoylphosphatidylglycerol and dipalmitoylphosphatidylcholine
-
-
?
additional information
?
-
enzyme diplays lipid-mediated unfolding. As isoform 3betaHSD2 lacks a receptor, opening the conformation may activate the protein by forming a cavity forms between the alpha-helix and beta-sheet
-
-
?
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
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
DELTA5-3beta-hydroxysteroid + NAD+
DELTA4-3-oxosteroid + NADH + H+
-
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
-
-
?
pregnenolone + NAD(P)+
pregn-5-en-3,20-dione + NAD(P)H + H+
-
-
-
-
?
17alpha-hydroxypregnenolone + NAD+
17alpha-hydroxy-pregn-5-en-3,20-dione + NADH
-
-
-
?
17alpha-hydroxypregnenolone + NAD+
17alpha-hydroxy-pregn-5-en-3,20-dione + NADH
-
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
?
dehydroepiandrosterone + NAD+
androst-5-en-3,17-dione + NADH + H+
-
-
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones catalyzing the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
-
-
?
additional information
?
-
-
the enzyme is essential for biosynthesis of all classes of steroid hormones catalyzing the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
-
-
?
additional information
?
-
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
the enzyme is essential for biosynthesis of all classes of steroid hormones, pathway overview
-
-
?
additional information
?
-
-
the enzyme is involved in steroid production and/or metabolism
-
-
?
additional information
?
-
using fluorescence resonance energy transfer, it is shown that 3betaHSD2 interacts strongly with a mixture of dipalmitoylphosphatidylglycerol and dipalmitoylphosphatidylcholine
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADP+
NADPH forms hydrogen bond with Lys36 in the NADPH-wild-type 3beta-HSD1 complex. The total contribution of Asp35 and Lys36 is most favorable in the case of NADPH binding to the wild-type enzyme when compared with NADH, and Lys36 have strong electronic interaction with NADPH
NAD+
NADH forms hydrogen bond with Asp35 in the NADH-wild-type 3beta-HSD1 complex
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
16-cyano-17-keto-trilostane
i.e 2,16-dicyano-4,5-epoxy-androstane-3,17-dione. Competitive, selective inhibitor of wild-type enzyme and chimeric S194G-1 mutant of the enzyme (3beta-HSD1). Noncompetitive inhibitor of chimeric R195P-1 mutant of the enzyme. Selective inhibition of 3beta-HSD1 may prevent spontaneous premature birth
-
organotin
can inhibit placental HSD3B1 activity, thus blocking the formation of progesterone and disrupting pregnancy
-
2,16-dicyano-4,5-epoxy-androstane-3,17-dione
competitively inhibits 3beta-HSD1
-
epostane
inhibits the 3beta-HSD activity of the wild-type 1 isozyme with 1417fold greater affinity compared to the wild-type 2 isozyme and mutant H156Y
epostane
inhibits the dehydroepiandrosterone-induced proliferation of MCF-7 cells expressing isoform 3beta-HSD1 with 12- to 16fold lower IC50 values compared to the MCF-7 cells expressing isoform 3beta-HSD2. Epostane also competitively inhibits purified human 3beta-HSD1 with 12fold to 16fold lower Ki values compared to the noncompetitive Ki values measured for human 3beta-HSD2
trilostane
inhibits the dehydroepiandrosterone-induced proliferation of MCF-7 cells expressing isoform 3beta-HSD1 with 12- to 16fold lower IC50 values compared to the MCF-7 cells expressing isoform 3beta-HSD2. Trilostane also competitively inhibits purified human 3beta-HSD1 with 12fold to 16fold lower Ki values compared to the noncompetitive Ki values measured for human 3beta-HSD2. Trilostane is docked in the active site of isoform 3beta-HSD1, and residue Arg195 in 3beta-HSD1 is a potentially critical residue
trilostane
noncompetitive inhibitor of chimeric S194G-1 mutant and chimeric R195P-1 mutant of the enzyme (3beta-HSD1). Competitive inhibition of wild-type enzyme
16-cyano-17-keto-trilostane
i.e 2,16-dicyano-4,5-epoxy-androstane-3,17-dione. noncompetitive inhibitor
-
epostane
inhibits the 3beta-HSD activity of the wild-type 1 isozyme with 1417fold greater affinity compared to the wild-type 2 isozyme and mutant H156Y
epostane
inhibits the dehydroepiandrosterone-induced proliferation of MCF-7 cells expressing isoform 3beta-HSD1 with 12- to 16fold lower IC50 values compared to the MCF-7 cells expressing isoform 3beta-HSD. Epostane also competitively inhibits purified human 3beta-HSD1 with 12fold to 16fold lower Ki values compared to the noncompetitive Ki values measured for human 3beta-HSD2
trilostane
-
i.e. 2alpha-cyano-4alpha,5alpha-epoxy-17beta-ol-androstane-3-one, competitive inhibition
trilostane
-
human adrenal H295R cells treated with the HSD3B2 inhibitor, trilostane, demonstrate a markedly decreased androstenedione production
trilostane
inhibits the dehydroepiandrosterone-induced proliferation of MCF-7 cells expressing isoform 3beta-HSD1 with 12- to 16fold lower IC50 values compared to the MCF-7 cells expressing isoform 3beta-HSD2. Trilostane also competitively inhibits purified human 3beta-HSD1 with 12fold to 16fold lower Ki values compared to the noncompetitive Ki values measured for human 3beta-HSD2
additional information
-
no inhibition by quercetin, apigenin, genistein, daidzein, enterodiol, and enterolactone
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0064
16alpha-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD1, pH 7.4, 27°C
0.0196
16alpha-hydroxy-dehydroepiandrosterone
mutant R195P, pH 7.4, 27°C
0.0014
16beta-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD1, pH 7.4, 27°C
0.015
16beta-hydroxy-dehydroepiandrosterone
mutant R195P, pH 7.4, 27°C
0.0037
dehydroepiandrosterone
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD1
0.0037
dehydroepiandrosterone
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD1
0.0061
dehydroepiandrosterone
pH 7.4, 27°C, recombinant isozyme 3beta-HSD1 mutant Q240R
0.0107
dehydroepiandrosterone
pH 7.4, 27°C, chimeric R195P-1 mutant of the enzyme (3beta-HSD1)
0.0124
dehydroepiandrosterone
pH 7.4, 27°C, chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
0.0405
dehydroepiandrosterone
pH 7.4, 27°C, recombinant isozyme 3beta-HSD1 mutant Q105M
0.0424
dehydroepiandrosterone
pH 7.4, 27°C, recombinant mutant H156Y
0.0244
NAD+
pH 7.4, 27°C, chimeric R195P-1 mutant of the enzyme (3beta-HSD1)
0.0256
NAD+
pH 7.4, 27°C, recombinant isozyme 3beta-HSD1 mutant Q240R
0.0341
NAD+
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD1
0.0341
NAD+
pH 7.4, 27°C, chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
0.0388
NAD+
pH 7.4, 27°C, recombinant isozyme 3beta-HSD1 mutant Q105M
0.004
16alpha-hydroxy-dehydroepiandrosterone
mutant R195R, pH 7.4, 27°C
0.0148
16alpha-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD2, pH 7.4, 27°C
0.0015
16beta-hydroxy-dehydroepiandrosterone
mutant R195R, pH 7.4, 27°C
0.0184
16beta-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD2, pH 7.4, 27°C
0.0044
dehydroepiandrosterone
-
mutant C72S, presence of 2-mercatoethanol pH 7.4
0.0075
dehydroepiandrosterone
-
mutant C83A, presence of 2-mercatoethanol pH 7.4
0.0115
dehydroepiandrosterone
-
mutant C72F, presence of 2-mercatoethanol pH 7.4
0.0168
dehydroepiandrosterone
-
mutant C111S, presence of 2-mercatoethanol pH 7.4
0.0237
dehydroepiandrosterone
-
mutant C111A, presence of 2-mercatoethanol pH 7.4
0.025
dehydroepiandrosterone
pH 7.4, 27°C, recombinant isozyme 3beta-HSD2 mutant Q105M
0.0327
dehydroepiandrosterone
pH 7.4, 27°C, recombinant isozyme 3beta-HSD2 mutant R240Q
0.042
dehydroepiandrosterone
27°C, pH 7.4, H156Y mutant 3beta-HSD1
0.0424
dehydroepiandrosterone
pH 7.4, 27°C, recombinant mutant H156Y
0.047
dehydroepiandrosterone
pH and temperature not specified in the publication
0.0473
dehydroepiandrosterone
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD2
0.0473
dehydroepiandrosterone
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD2
0.0372
NAD+
pH 7.4, 27°C, recombinant isozyme 3beta-HSD2 mutant R240Q
0.0614
NAD+
pH 7.4, 27°C, recombinant isozyme 3beta-HSD2 mutant Q105M
0.0863
NAD+
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD2
additional information
additional information
-
kinetics recombinant wild-type isozymes and mutant H165Y, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
kinetics recombinant wild-type isozymes and mutant H165Y, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
kinetics recombinant wild-type isozymes and mutant H165Y, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
kinetics, recombinant wild-type and mutant isozymes, overview, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
kinetics, recombinant wild-type and mutant isozymes, overview, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
-
kinetics, recombinant wild-type and mutant isozymes, overview, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
-
kinetics recombinant wild-type isozyme and mutant H165Y, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
kinetics recombinant wild-type isozyme and mutant H165Y, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
kinetics recombinant wild-type isozyme and mutant H165Y, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
kinetics, recombinant wild-type and mutant isozymes, overview, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
kinetics, recombinant wild-type and mutant isozymes, overview, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
additional information
additional information
-
kinetics, recombinant wild-type and mutant isozymes, overview, kinetics for the DELTA-5-3-ketosteroid isomerase, EC 5.3.3.1, activity
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.09
16alpha-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD1, pH 7.4, 27°C
0.15
16alpha-hydroxy-dehydroepiandrosterone
mutant R195P, pH 7.4, 27°C
0.04
16beta-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD1, pH 7.4, 27°C
0.15
16beta-hydroxy-dehydroepiandrosterone
mutant R195P, pH 7.4, 27°C
0.023
dehydroepiandrosterone
pH 7.4, 27°C, recombinant isozyme 3beta-HSD1 mutant Q240R
0.055
dehydroepiandrosterone
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD1
0.062
dehydroepiandrosterone
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD1
0.113
dehydroepiandrosterone
pH 7.4, 27°C, recombinant isozyme 3beta-HSD1 mutant Q105M
0.12
dehydroepiandrosterone
pH 7.4, 27°C, recombinant mutant H156Y
0.128
dehydroepiandrosterone
pH 7.4, 27°C, chimeric R195P-1 mutant of the enzyme (3beta-HSD1)
0.13
dehydroepiandrosterone
pH 7.4, 27°C, chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
0.022
NAD+
pH 7.4, 27°C, recombinant isozyme 3beta-HSD1 mutant Q240R
0.038
NAD+
pH 7.4, 27°C, recombinant isozyme 3beta-HSD1 mutant Q105M
0.04
NAD+
pH 7.4, 27°C, chimeric R195P-1 mutant of the enzyme (3beta-HSD1)
0.06
NAD+
pH 7.4, 27°C, chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
0.07
16alpha-hydroxy-dehydroepiandrosterone
mutant R195R, pH 7.4, 27°C
0.15
16alpha-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD2, pH 7.4, 27°C
0.05
16beta-hydroxy-dehydroepiandrosterone
mutant R195R, pH 7.4, 27°C
0.14
16beta-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD2, pH 7.4, 27°C
0.047
dehydroepiandrosterone
-
mutant C111A, presence of 2-mercatoethanol pH 7.4
0.075
dehydroepiandrosterone
pH 7.4, 27°C, recombinant isozyme 3beta-HSD2 mutant Q105M
0.115
dehydroepiandrosterone
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD2
0.118
dehydroepiandrosterone
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD2
0.12
dehydroepiandrosterone
pH 7.4, 27°C, recombinant mutant H156Y
0.217
dehydroepiandrosterone
pH 7.4, 27°C, recombinant isozyme 3beta-HSD2 mutant R240Q
0.145
NAD+
pH 7.4, 27°C, recombinant isozyme 3beta-HSD2 mutant Q105M
0.187
NAD+
pH 7.4, 27°C, recombinant isozyme 3beta-HSD2 mutant R240Q
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
7.83
16alpha-hydroxy-dehydroepiandrosterone
mutant R195P, pH 7.4, 27°C
15
16alpha-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD1, pH 7.4, 27°C
10.33
16beta-hydroxy-dehydroepiandrosterone
mutant R195P, pH 7.4, 27°C
34.5
16beta-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD1, pH 7.4, 27°C
10.48
dehydroepiandrosterone
pH 7.4, 27°C, chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
11.96
dehydroepiandrosterone
pH 7.4, 27°C, chimeric R195P-1 mutant of the enzyme (3beta-HSD1)
0.176
NAD+
pH 7.4, 27°C, chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
1.64
NAD+
pH 7.4, 27°C, chimeric R195P-1 mutant of the enzyme (3beta-HSD1)
10.16
16alpha-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD2, pH 7.4, 27°C
19.1
16alpha-hydroxy-dehydroepiandrosterone
mutant R195R, pH 7.4, 27°C
7.66
16beta-hydroxy-dehydroepiandrosterone
wild-type 3beta-HSD2, pH 7.4, 27°C
35.5
16beta-hydroxy-dehydroepiandrosterone
mutant R195R, pH 7.4, 27°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0016
16-cyano-17-keto-trilostane
pH 7.4, 27°C, wild-type enzyme
-
0.00043
17beta-acetoxy-trilostane
-
in 0.02 M potassium phosphate buffer, pH 7.4
0.0307
2,16-dicyano-4,5-epoxy-androstane-3,17-dione
pH 7.4, 27°C, wild-type enzyme
-
0.000029
nomegestrol acetate
-
pH and temperature not specified in the publication
0.0047
16-cyano-17-keto-trilostane
pH 7.4, 27°C, wild-type enzyme
-
0.0295
16-cyano-17-keto-trilostane
pH 7.4, 27°C, chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
-
0.0413
16-cyano-17-keto-trilostane
pH 7.4, 27°C, chimeric R195P-1 mutant of the enzyme (3beta-HSD1)
-
0.000071
epostane
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD1
0.00067
trilostane
pH 7.4, 27°C, chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
0.00256
trilostane
pH 7.4, 27°C, chimeric R195P-1 mutant of the enzyme (3beta-HSD1)
0.0028
4alpha,5alpha-epoxy-testosterone
-
mutant enzyme M187T, in 0.02 M potassium phosphate, pH 7.4, at 27°C
0.00375
4alpha,5alpha-epoxy-testosterone
-
wild type enzyme 3beta-HSD2, in 0.02 M potassium phosphate, pH 7.4, at 27°C
0.00408
4alpha,5alpha-epoxy-testosterone
-
wild type enzyme 3beta-HSD1, in 0.02 M potassium phosphate, pH 7.4, at 27°C
0.00434
4alpha,5alpha-epoxy-testosterone
-
mutant enzyme S124T, in 0.02 M potassium phosphate, pH 7.4, at 27°C
0.00098
epostane
pH 7.4, 27°C, recombinant wild-type isozyme 3beta-HSD2
0.0001
trilostane
-
wild type enzyme 3beta-HSD1, in 0.02 M potassium phosphate, pH 7.4, at 27°C
0.00041
trilostane
-
mutant enzyme S124T, in 0.02 M potassium phosphate, pH 7.4, at 27°C
0.00083
trilostane
-
mutant enzyme M187T, in 0.02 M potassium phosphate, pH 7.4, at 27°C
0.0016
trilostane
-
wild type enzyme 3beta-HSD2, in 0.02 M potassium phosphate, pH 7.4, at 27°C
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.4
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
7.4
9.7
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
LITERATURE
27
27
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
abdominal, expression level determination, especially in female aorta with mild atherosclerotic changes
-
induction by peroxisome-proliferator-activated receptor alpha ligands
HSD3B mRNA levels in oocytes producing euploid embryos are significantly lower than other oocyte outcomes
-
in amnion epithelium immunoreactive 3betaHSD is found from first to third trimester and term
-
HSD3B2 and CYB5A are immonolocalized in normal human adrenal glands. Their co-expression is demonstrated in the cortical cells located at the border between the zona fasciculata and zona reticularis in normal human adrenal
3beta-HSD21 is selectively expressed in adrenal glands and ovaries
3beta-HSD21 is selectively expressed in adrenal glands and ovaries
-
in syncytiotrophoblast, cytotrophoblast and intermediate trophoblast immunoreactive 3betaHSD is found from first to third trimester and term
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
in amnion epithelium, chorion trophoblast and invasive trophoblast immunoreactive 3betaHSD is found from first to third trimester and term
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug target
16-cyano-17-keto-trilostane is a selective inhibitor of 3beta-hydroxysteroid dehydrogenase type 1. Selective inhibition of beta-HSD1 may prevent spontaneous premature birth. Inhibition of 3beta-HSD1 decreases placental estradiol production that promotes labor. The 3beta-HSD1 inhibitor may be administered with 17alpha-hydroxyprogesterone caproate
metabolism
HSD3B1 is a unique enzyme for progesterone synthesis and is also involved in the estrogen production in human placenta
drug target
16-cyano-17-keto-trilostane is a selective inhibitor of 3beta-hydroxysteroid dehydrogenase type 1. Selective inhibition of beta-HSD1 may prevent spontaneous premature birth. Inhibition of 3beta-HSD1 decreases placental estradiol production that promotes labor. The 3beta-HSD1 inhibitor may be administered with 17alpha-hydroxyprogesterone caproate
metabolism
metabolism
-
isoform 3betaHSD2 interacts with the mitochondrial translocase, Tim50, to regulate progesterone and androstenedione formation
metabolism
-
isoform 3bHSD2 interacts with the mitochondrial translocase, Tim50, to regulate progesterone and androstenedione formation
metabolism
isoform HSD3B1 is involved in aldosterone production in aldosterone-producing adenoma
metabolism
the enzyme is responsible for intermediate steroidogenic reaction for the synthesis of androgens, estrogens, progestins, glucocorticoids, and mineralocorticoids in the adrenal cortex, gonads, placenta, and a variety of peripheral target tissues
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). 3BHS1_HUMAN
373
1
42252
Swiss-Prot
other Location (Reliability: 5)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
42000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
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
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C83A
mutation produces fully active enzyme with elevated Km values for ligands
C83S
mutation in the active site abolishes the dehydrogenase and isomerase activities of the enzyme
D35A/K36R
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
E192A
the E192A mutation causes an 8.2fold increase in Km (dehydroepiandrosterone) and no change in the Km for NAD+
H156Y
K158Q
K273Q
site-directed mutagenesis of isozyme 1, inactive mutant showing no 3beta-hydroxysteroid dehydrogenase activity, but reduced isomerase activity
Q105M
site-directed mutagenesis of isozyme 1, the mutants show altered kinetics compared to the wild-type isozymes
Q240R
site-directed mutagenesis of isozyme 1, the mutant shows altered kinetics compared to the wild-type isozyme
R195P
S194G
16-cyano-17-keto-trilostane and trilostane inhibits the chimeric S194G-1 mutant of the enzyme (3beta-HSD1) with a noncompetitive mode compared to competitive inhibition of the wild-type enzyme (3beta-HSD1)
Y154F
Y269S
site-directed mutagenesis of isozyme 1, inactive mutant showing no 3beta-hydroxysteroid dehydrogenase activity, but reduced isomerase activity
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+
C83S
H156Y
K158Q
K273Q
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
Q105M
site-directed mutagenesis of isozyme 2, the mutants show altered kinetics compared to the wild-type isozymes
Q105M1
mutant retains the lower affinity profile of inhibition by epostane that is observed with wild-type 3beta-HSD2
R195P
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
S194G
16-cyano-17-keto-trilostane and trilostane inhibits the chimeric S194G-1 mutant of the enzyme (3beta-HSD1) with a noncompetitive mode compared to competitive inhibition of the wild-type enzyme (3beta-HSD1)
T177N
-
significantly improved stability at cold temperature, presence of NAD+ is required for complete stability at 0°C
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
Y269S
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
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
D35A/K36R
double mutations cause conversion of the cofactor dependence from NAD+ to NADP+
H156Y
site-directed mutagenesis of isozyme 1, construction of a chimeric Y154-P-H156-S-K158 motif of isozymes 1 and 2, the mutant shows altered kinetics compared to the wild-type isozyme 1, the kinetics are similar to those of isozyme 2
K158Q
site-directed mutagenesis of isozyme 1, inactive mutant showing no 3beta-hydroxysteroid dehydrogenase activity, but reduced isomerase activity
R195P
mutation of isoform 3betaHSD1 to corresponding residue in isoform 3betaHSD2. Mutation shifts the dehydroepiandrosterone substrate kinetics to the lower affinity profile of isoform 3beta-HSD2
R195P
R195P mutation shifts the high-affinity, competitive inhibition profile of 3beta-HSD1 to a low-affinity (Ki= 2.56 microM), noncompetitive inhibition profile. R195P mutant has a 10.4fold higher Km value for 16beta-hydroxy-dehydroepiandrosterone compared to wild-type 3beta-HSD1. Km values for 16alpha-hydroxy-dehydroepiandrosterone and dehydroepiandrosterone are also inreased compared to wild-type (kcat values are decreased compared to wild-type)
R195P
16-cyano-17-keto-trilostane inhibits the chimeric R195P-1 mutant of the enzyme (3beta-HSD1) with a noncompetitive mode compared to competitive inhibition of the wild-type enzyme (3beta-HSD1)
Y154F
site-directed mutagenesis of isozyme 1, inactive mutant showing no 3beta-hydroxysteroid dehydrogenase activity, but reduced isomerase activity
C83S
-
similar to wild-type, presence of 2-mercaptoethanol results in 2.5fold decrease of Km values
H156Y
site-directed mutagenesis of isozyme 1, construction of a chimeric Y154-P-H156-S-K158 motif of isozymes 1 and 2, the mutant shows altered kinetics compared to the wild-type isozyme 1, the kinetics are similar to those of isozyme 2
R195P
mutation of isoform 3betaHSD2 to corresponding residue in isoform 3betaHSD1. Mutation shifts the dehydroepiandrosterone substrate kinetics to the higher affinity profile of isoform 3beta-HSD1
R195P
16-cyano-17-keto-trilostane inhibits the chimeric R195P-1 mutant of the enzyme (3beta-HSD1) with a noncompetitive mode compared to competitive inhibition of the wild-type enzyme (3beta-HSD1)
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.5 - 7.4
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
725511
4.5 - 7.4
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
LITERATURE
37
42
37
3betaHSD2 becomes less stable when incubated with the individual lipids dipalmitoylphosphatidylglycerol or dipalmitoylphosphatidylcholine, as indicated by the decrease in thermal denaturation (Tm) from 42 to 37°C. Dipalmitoylphosphatidylglycerol alone or in combination with dipalmitoylphosphatidylcholine, leads to a decrease in alpha-helical content without an effect on the beta-sheet conformation
37
approximately, melting temperature in the presence of zwitterionically charged dipalmitoylphosphatidylcholine vesicles or an equal mixture of zwitterionic vesicles and anionic vesicles
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purification of the chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
recombinant soluble wild-type and mutant isozyme 3beta-HSD1 from Sf9 insect cells
purification of the chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
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 soluble wild-type and mutant isozyme 3beta-HSD2 from Sf9 insect cells
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of soluble wild-type and mutant isozyme 3beta-HSD1 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system
expression of the chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
expressed in Sf9 cells using the baculovirus system
expression in Sf9 cells
expression of soluble wild-type and mutant isozyme 3beta-HSD2 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system
expression of the chimeric S194G-1 mutant of the enzyme (3beta-HSD1)
expression of the isozyme type II in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system
-
gene HSD3B2, analysis of expression and its regulation involving Nur77 factors in H295R and MA-10 cell lines, overview
-
overexpression with his6-tag, using a baculovirus expression system
-
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-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 of soluble wild-type and mutant isozyme 3beta-HSD2 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression is regulated by GATA2 and TEF-5 transcription factors. GATA2 is the positively regulatory transcription factor
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
HSD3B mRNA levels in oocytes producing euploid embryos are significantly lower than other oocyte outcomes
the mRNA level of the enzyme is higher in tumor tissue compared with those of the normal tissue
-
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
LITERATURE
diagnostics
HSD3B1 mRNA levels may be able to identify oocytes capable of producing euploid embryos
medicine
medicine
-
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
medicine
-
identification of mutations L341P, W355stop, and R355stop, in HSD3B2 gene in neonates with classical 3beta-hydroxysteroid dehydrogenase type II deficiency and under-virilization
medicine
-
patient with inborn error of bile acid metabolism, that is, 3beta-hydroxy-DELTA5-C27-steroid dehydrogenase/isomerase deficienc, receives as treatment chenodeoxycholic acid and cholic acid, orally administered for 10 years. After the oral bile acid therapy, the biochemical liver function values and serum alanine aminotransferase and total bilirubin levels are maintained at normal levels, while the 3beta,7alpha-dihydroxy- and 3beta,7alpha,12alpha-trihydroxy-5-cholenoic acid levels significantly decrease. The patient gave birth to a healthy boy and a girl during the 10-year follow-up period
medicine
-
in a female patient with pseudo-hermaphrodism caused by an androgen-producing adrenocortical tumor. The activities of 3beta-hydroxysteroid dehydrogenase 2 and CYP17 in the tumor tissue are higher than those in the normal tissue, whereas the activity of 17beta?hydroxysteroid dehydrogenase 3 is lower. The mRNA levels of 3beta-hydroxysteroid dehydrogenase 2 and CYP17 are higher and the levels of 17beta-hydroxysteroid dehydrogenase 3 and LH/hCG receptor are lower in the tumor tissue compared with those of the normal tissue
medicine
in aldosterone-producing adenoma, isoform HSD3B2 mRNA is significantly more abundant than HSD3B1 mRNA, but only HSD3B1 mRNA significantly correlates with CYP11B2 (aldosterone synthase) mRNA and plasma aldosterone concentration of the patients. A significant correlation is also detected between isoform HSD3B1 and CYP11B2. In KCNJ5 mutated aldosterone-producing adenoma, CYP11B2 mRNA and HSD3B1 mRNA are significantly higher than those of wild-type aldosterone-producing adenoma
medicine
in aldosterone-producing adenoma, isoform HSD3B2 mRNA is significantly more abundant than HSD3B1 mRNA, but only HSD3B1 mRNA significantly correlates with CYP11B2 (aldosterone synthase) mRNA and plasma aldosterone concentration of the patients. Isoform HSD3B2 immunoreactivity is detected in the great majority of aldosterone-producing adenoma
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Thomas, J.L.; Strickler, R.C.; Myers, R.P.; Covey, D.F
Affinity labeling of human placental 3beta-hydroxy-DELTA5-steroid dehydrogenase and steroid delta-isomerase: evidence for bifunctional catalysis by a different conformation of the same protein for each enzyme activity
Biochemistry
31
5522-5527
1992
Homo sapiens
Rabe, T.; Brandstetter, K.; Kellermann, J.; Runnebaum, B.
Partial characterization of placental 3beta-hydroxysteroid dehydrogenase (EC 1.1.1.145), delta 4-5isomerase (EC 5.3.3.1) in human term placental mitochondria
J. Steroid Biochem.
17
427-433
1982
Homo sapiens
Huang, Y.W.; Lu, M.L.; Qi, H.; Lin, S.X.
Membrane-bound human 3beta-hydroxysteroid dehydrogenase: overexpression with his-tag using a baculvirus system and single-step purification
Protein Expr. Purif.
18
169-174
2000
Homo sapiens
Thomas, J.L.; Mason, J.I.; Blanco, G.; Veisaga, M.L.
The engineered, cytosolic form of human type I 3beta-hydroxysteroid dehydrogenase/isomerase: purification, characterization and crystallization
J. Mol. Endocrinol.
27
77-83
2001
Homo sapiens
Thomas, J.L.; Evans, B.W.; Blanco, G.; Mason, J.I.; Strickler, R.C.
Creation of a fully active, cytosolic form of human type I 3beta-hydroxysteroid dehydrogenase/isomerase by the deletion of a membrane-spanning domain
J. Mol. Endocrinol.
23
231-239
1999
Homo sapiens
Riley, S.C.; Dupont, E.; Walton, J.C.; Luu-The, V.; Labrie, F.; Pelletier, G.; Challis, J.R.
Immunohistochemical localization of 3beta-hydroxy-5-ene-steroid dehydrogenase/DELTA5-DELTA4 isomerase in human placenta and fetal membranes throughout gestation
J. Clin. Endocrinol.
75
956-961
1992
Homo sapiens
Suzuki, T.; Sasana, H.; Andersson, S.; Mason, J.I.
3beta-Hydroxysteroid dehydrogenase/DELTA5---4-isomerase activity associated with the human 17beta-hydroxysteroid dehydrogenase type 2 isoform
J. Clin. Endocrinol.
85
3669-3672
2000
Homo sapiens
Labrie, F.; Simard, J.; Luu-The, V.; Pelletier, G.; Belanger, A.; Lachance, Y.; Zhao, H.F.; Labrie, C.; Breton, N.; de Launoit, Y.; et al.
Structure and tissue-specific expression of 3beta-hydroxysteroid dehydrogenase/5-ene-4-ene isomerase genes in human and rat classical and peripheral steroidogenic tissues
J. Steroid Biochem. Mol. Biol.
41
421-435
1992
Homo sapiens, Rattus norvegicus
Nash, W.E.; Mercer, R.W.; Blanco, G.; Strickler, R.C.; Mason, J.I.; Thomas, J.L.
Over-expression of human type I (placental) 3beta-hydroxy-5-ene-steroid dehydrogenase/isomerase in insect cells infected with recombinant baculovirus
J. Steroid Biochem. Mol. Biol.
50
235-240
1994
Homo sapiens
Thomas, J.L.; Mason, J.I.; Brandt, S.; Norris, W.
Differences in substrate and inhibitor kinetics of human type 1 and type 2 3beta-hydroxysteroid dehydrogenase are explained by the type 1 mutant, H156Y
Endocr. Res.
28
471-475
2002
Homo sapiens
Thomas, J.L.; Mason, J.I.; Brandt, S.; Spencer, B.R., Jr.; Norris, W.
Structure/function relationships responsible for the kinetic differences between human type 1 and type 2 3beta-hydroxysteroid dehydrogenase and for the catalysis of the type 1 activity
J. Biol. Chem.
277
42795-42801
2002
Homo sapiens, Homo sapiens (P14060), Homo sapiens (P26439)
Thomas, J.L.; Duax, W.L.; Addlagatta, A.; Scaccia, L.A.; Frizzell, K.A.; Carloni, S.B.
Serine 124 completes the Tyr, Lys and Ser triad responsible for the catalysis of human type 1 3beta-hydroxysteroid dehydrogenase
J. Mol. Endocrinol.
33
253-261
2004
Homo sapiens
Ohno, S.; Matsumoto, N.; Watanabe, M.; Nakajin, S.
Flavonoid inhibition of overexpressed human 3beta-hydroxysteroid dehydrogenase type II
J. Steroid Biochem. Mol. Biol.
88
175-182
2004
Homo sapiens
Thomas, J.L.; Duax, W.L.; Addlagatta, A.; Kacsoh, B.; Brandt, S.E.; Norris, W.B.
Structure/function aspects of human 3beta-hydroxysteroid dehydrogenase
Mol. Cell. Endocrinol.
215
73-82
2004
Homo sapiens
Thomas, J.L.; Boswell, E.L.; Scaccia, L.A.; Pletnev, V.; Umland, T.C.
Identification of key amino acids responsible for the substantially higher affinities of human type 1 3beta-hydroxysteroid dehydrogenase/isomerase (3b-HSD1) for substrates, coenzymes, and inhibitors relative to human 3b-HSD2
J. Biol. Chem.
280
21321-21328
2005
Homo sapiens (P14060), Homo sapiens (P26439), Homo sapiens
Nakamura, Y.; Suzuki, T.; Inoue, T.; Tazawa, C.; Moriya, T.; Saito, H.; Ishibashi, T.; Takahashi, S.; Yamada, S.; Sasano, H.
3beta-hydroxysteroid dehydrogenase in human aorta
Endocr. J.
52
111-115
2005
Homo sapiens
Martin, L.J.; Tremblay, J.J.
The human 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase type 2 promoter is a novel target for the immediate early orphan nuclear receptor Nur77 in steroidogenic cells
Endocrinology
146
861-869
2005
Homo sapiens
Pletnev, V.Z.; Thomas, J.L.; Rhaney, F.L.; Holt, L.S.; Scaccia, L.A.; Umland, T.C.; Duax, W.L.
Rational proteomics V: structure-based mutagenesis has revealed key residues responsible for substrate recognition and catalysis by the dehydrogenase and isomerase activities in human 3beta-hydroxysteroid dehydrogenase/isomerase type 1
J. Steroid Biochem. Mol. Biol.
101
50-60
2006
Homo sapiens (P14060), Homo sapiens
Lacey, M.; Bohday, J.; Fonseka, S.M.; Ullah, A.I.; Whitehead, S.A.
Dose-response effects of phytoestrogens on the activity and expression of 3beta-hydroxysteroid dehydrogenase and aromatase in human granulosa-luteal cells
J. Steroid Biochem. Mol. Biol.
96
279-286
2005
Homo sapiens
Matsunaga, T.; Endo, S.; Maeda, S.; Ishikura, S.; Tajima, K.; Tanaka, N.; Nakamura, K.T.; Imamura, Y.; Hara, A.
Characterization of human DHRS4: An inducible short-chain dehydrogenase/reductase enzyme with 3beta-hydroxysteroid dehydrogenase activity
Arch. Biochem. Biophys.
477
339-347
2008
Homo sapiens
Welzel, M.; Wuestemann, N.; Simic-Schleicher, G.; Doerr, H.G.; Schulze, E.; Shaikh, G.; Clayton, P.; Groetzinger, J.; Holterhus, P.M.; Riepe, F.G.
Carboxyl-terminal mutations in 3beta-hydroxysteroid dehydrogenase type II cause severe salt-wasting congenital adrenal hyperplasia
J. Clin. Endocrinol. Metab.
93
1418-1425
2008
Homo sapiens
Thomas, J.L.; Huether, R.; Mack, V.L.; Scaccia, L.A.; Stoner, R.C.; Duax, W.L.
Structure/function of human type 1 3beta-hydroxysteroid dehydrogenase: An intrasubunit disulfide bond in the Rossmann-fold domain and a Cys residue in the active site are critical for substrate and coenzyme utilization
J. Steroid Biochem. Mol. Biol.
107
80-87
2007
Homo sapiens
Wang, L.; Salavaggione, E.; Pelleymounter, L.; Eckloff, B.; Wieben, E.; Weinshilboum, R.
Human 3beta-hydroxysteroid dehydrogenase types 1 and 2: Gene sequence variation and functional genomics
J. Steroid Biochem. Mol. Biol.
107
88-99
2007
Homo sapiens
Thomas, J.L.; Mack, V.L.; Glow, J.A.; Moshkelani, D.; Terrell, J.R.; Bucholtz, K.M.
Structure/function of the inhibition of human 3beta-hydroxysteroid dehydrogenase type 1 and type 2 by trilostane
J. Steroid Biochem. Mol. Biol.
111
66-73
2008
Homo sapiens
Hu, G.X.; Zhou, H.Y.; Li, X.W.; Chen, B.B.; Xiao, Y.C.; Lian, Q.Q.; Liang, G.; Kim, H.H.; Zheng, Z.Q.; Hardy, D.O.; Ge, R.S.
The (+)- and (-)-gossypols potently inhibit both 3beta-hydroxysteroid dehydrogenase and 17beta-hydroxysteroid dehydrogenase 3 in human and rat testes
J. Steroid Biochem. Mol. Biol.
115
14-19
2009
Homo sapiens, Rattus norvegicus
Thomas, J.L.; Bucholtz, K.M.; Sun, J.; Mack, V.L.; Kacsoh, B.
Structural basis for the selective inhibition of human 3beta-hydroxysteroid dehydrogenase 1 in human breast tumor MCF-7 cells
Mol. Cell. Endocrinol.
301
174-182
2009
Homo sapiens
Nittono, H.; Takei, H.; Unno, A.; Shimizu, T.; Kobayashi, M.; Koike, M.; Murai, T.; Kurosawa, T.; Thoma, M.
3?-hydroxy-??-C??-steroid dehydrogenase/isomerase deficiency in a patient who underwent oral bile acid therapy for 10 years and delivered two healthy infants.
Pediatr. Internat.
52
e192-e195
2010
Homo sapiens
Rajapaksha, M.; Thomas, J.L.; Streeter, M.; Prasad, M.; Whittal, R.M.; Bell, J.D.; Bose, H.S.
Lipid-mediated unfolding of 3beta-hydroxysteroid dehydrogenase 2 is essential for steroidogenic activity
Biochemistry
50
11015-11024
2011
Homo sapiens (P26439)
Prasad, M.; Thomas, J.L.; Whittal, R.M.; Bose, H.S.
Mitochondrial 3beta-hydroxysteroid dehydrogenase enzyme activity requires reversible pH-dependent conformational change at the intermembrane space
J. Biol. Chem.
287
9534-9546
2012
Homo sapiens (P26439)
Zhao, Y.; Xiao, J.
Homology modeling and molecular dynamics simulation studies of human type 1 3beta-hydroxysteroid dehydrogenase: Toward the understanding of cofactor specificity
J. Comput. Chem.
32
33-42
2011
Homo sapiens (P14060), Homo sapiens
Thomas, J.L.; Mack, V.L.; Sun, J.; Terrell, J.R.; Bucholtz, K.M.
The functions of key residues in the inhibitor, substrate and cofactor sites of human 3beta-hydroxysteroid dehydrogenase type 1 are validated by mutagenesis
J. Steroid Biochem. Mol. Biol.
120
192-199
2010
Homo sapiens (P14060), Homo sapiens (P26439)
Nakamura, Y.; Xing, Y.; Hui, X.G.; Kurotaki, Y.; Ono, K.; Cohen, T.; Sasano, H.; Rainey, W.E.
Human adrenal cells that express both 3?-hydroxysteroid dehydrogenase type 2 (HSD3B2) and cytochrome b5 (CYB5A) contribute to adrenal androstenedione production
J. Steroid Biochem. Mol. Biol.
123
122-126
2011
Homo sapiens
Thomas, J.; Bucholtz, K.; Kacsoh, B.
Selective inhibition of human 3beta-hydroxysteroid dehydrogenase type 1 as a potential treatment for breast cancer
J. Steroid Biochem. Mol. Biol.
125
57-65
2011
Homo sapiens (P14060), Homo sapiens (P26439)
Yang, G.; Dou, J.; Zhang, X.; Gu, W.; Lv, Z.; DU, J.; Ba, J.; Mu, Y.; Lu, J.
Increased 3beta-hydroxysteroid dehydrogenase 2 and 17beta-hydroxylase activities in a virilized adolescent female with adrenal adenoma: A case report
Exp. Ther. Med.
11
530-534
2016
Homo sapiens
Thomas, J.L.; Bose, H.S.
Regulation of human 3-beta-hydroxysteroid dehydrogenase type-2 (3betaHSD2) by molecular chaperones and the mitochondrial environment affects steroidogenesis
J. Steroid Biochem. Mol. Biol.
151
74-84
2015
Homo sapiens
Ota, T.; Doi, M.; Yamazaki, F.; Yarimizu, D.; Okada, K.; Murai, I.; Hayashi, H.; Kunisue, S.; Nakagawa, Y.; Okamura, H.
Angiotensin II triggers expression of the adrenal gland zona glomerulosa-specific 3beta-hydroxysteroid dehydrogenase isoenzyme through de novo protein synthesis of the orphan nuclear receptors NGFIB and NURR1
Mol. Cell. Biol.
34
3880-3894
2014
Homo sapiens (P14060), Homo sapiens
Konosu-Fukaya, S.; Nakamura, Y.; Satoh, F.; Felizola, S.J.; Maekawa, T.; Ono, Y.; Morimoto, R.; Ise, K.; Takeda, K.; Katsu, K.; Fujishima, F.; Kasajima, A.; Watanabe, M.; Arai, Y.; Gomez-Sanchez, E.P.; Gomez-Sanchez, C.E.; Doi, M.; Okamura, H.; Sasano, H.
3beta-Hydroxysteroid dehydrogenase isoforms in human aldosterone-producing adenoma
Mol. Cell. Endocrinol.
408
205-212
2015
Homo sapiens, Homo sapiens (P26439)
Louw-du Toit, R.; Perkins, M.S.; Snoep, J.L.; Storbeck, K.H.; Africander, D.
Fourth-generation progestins inhibit 3beta-hydroxysteroid dehydrogenase type 2 and modulate the biosynthesis of endogenous steroids
PLoS ONE
11
e0164170
2016
Homo sapiens
Ando, T.; Nishiyama, T.; Takizawa, I.; Ishizaki, F.; Miyashiro, Y.; Takeda, K.; Hara, N.; Tomita, Y.
Dihydrotestosterone synthesis pathways from inactive androgen 5alpha-androstane-3beta,17beta-diol in prostate cancer cells: Inhibition of intratumoural 3beta-hydroxysteroid dehydrogenase activities by abiraterone
Sci. Rep.
6
32198
2016
Homo sapiens
Zhang, S.; Mo, J.; Wang, Y.; Ni, C.; Li, X.; Zhu, Q.; Ge, R.S.
Endocrine disruptors of inhibiting testicular 3beta-hydroxysteroid dehydrogenase
Chem. Biol. Interact.
303
90-97
2019
Homo sapiens (P26439), Homo sapiens
Kordus, R.J.; Hossain, A.; Corso, M.C.; Chakraborty, H.; Whitman-Elia, G.F.; LaVoie, H.A.
Cumulus cell pappalysin-1, luteinizing hormone/choriogonadotropin receptor, amphiregulin and hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 mRNA levels associate with oocyte developmental competence and embryo outcomes
J. Assist. Reprod. Genet.
36
1457-1469
2019
Homo sapiens (P26439), Homo sapiens
Pham, J.H.; Will, C.M.; Mack, V.L.; Halbert, M.; Conner, E.A.; Bucholtz, K.M.; Thomas, J.L.
Structure-function relationships for the selective inhibition of human 3beta-hydroxysteroid dehydrogenase type 1 by a novel androgen analog
J. Steroid Biochem. Mol. Biol.
174
257-264
2017
Homo sapiens (P14060), Homo sapiens (P26439)
EXTERNAL LINKS (specific for EC-Number 1.1.1.145)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
