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Information on EC 5.3.3.1 - steroid DELTA-isomerase
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5.3.3.1 steroid DELTA-isomeraseIUBMB Comments
This activity is catalysed by several distinct enzymes (cf. EC 1.1.3.6, cholesterol oxidase and EC 1.1.1.145, 3-hydroxy-5-steroid dehydrogenase).
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Word Map
- 5.3.3.1
-
steroidogenic
- progesterone
- adrenal
-
steroidogenesis
- androgen
- p450scc
- leydig
- star
- testicular
- pregnenolone
- 17beta-hsd
- lh
- aromatase
-
5-3-ketosteroids
- gonad
- testosteroni
-
granulosa
- dehydroepiandrosterone
- luteal
- androstenedione
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17beta-hydroxysteroid
- dheas
-
theca
- p450arom
- 17alpha-hydroxylase
-
neurosteroids
- cyp11a1
- trilostane
- 19-nortestosterone
-
dienolate
-
5alpha-reductase
- equilenin
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low-barrier
- 17alpha-hydroxylase/17,20-lyase
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interna
- 3betahsd
- delta5-3beta-hydroxysteroid
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intracrine
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double-bond
- hsd3b1
-
talalay
- 20alpha-hsd
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beta-proton
The enzyme appears in viruses and cellular organisms
Synonyms
3beta-hsd, ketosteroid isomerase, delta 5-3-ketosteroid isomerase, steroid isomerase, 3beta-hydroxysteroid dehydrogenase/isomerase, steroid delta-isomerase, gst a3-3, 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase, 3beta-hsd/isomerase, 5-ene-4-ene isomerase, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
3-Keto-DELTA5-steroid isomerase
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3-Ketosteroid DELTA5-->DELTA4-isomerase
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3-Oxo steroid DELTA4-DELTA5-isomerase
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3-Oxo-delta5 steroid isomerase
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3-Oxosteroid DELTA4-DELTA5-isomerase
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3-Oxosteroid DELTA5-DELTA4-isomerase
3-Oxosteroid isomerase
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3beta-HSD
3beta-HSD/isomerase
5-Ene-4-ene isomerase
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5-Pregnene-3,20-dione isomerase
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delta 5-3-ketosteroid isomerase
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DELTA-3-ketosteroid isomerase
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Delta-5-3-ketosteroid isomerase
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DELTA5(or DELTA4)-3-keto steroid isomerase
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DELTA5-3-keto steroid isomerase
DELTA5-3-ketosteroid isomerase
DELTA5-3-oxosteroid isomerase
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DELTA5-ketosteroid isomerase
DELTA5-steroid isomerase
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Hydroxysteroid isomerase
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Isomerase, steroid DELTA
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ketosteroid isomerase
KSI
Steroid 5-->4-isomerase
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Steroid isomerase
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additional information
3-Oxosteroid DELTA5-DELTA4-isomerase
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DELTA5-3-keto steroid isomerase
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DELTA5-3-ketosteroid isomerase
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DELTA5-ketosteroid isomerase
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additional information
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3beta-hydroxy-5-ene steroid dehydrogenase/isomerase enzyme system
additional information
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3beta-hydroxy-DELTA5-steroid dehydrogenase/3-keto-DELTA5-steroid isomerase
additional information
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3beta-hydroxysteroid dehydrogenase/5-ene--4-ene isomerase
additional information
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beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase
additional information
the enzyme is a member of the short chain dehydrogenase/reductase family, cf. EC 1.1.1.51
additional information
the enzyme is a member of the short chain dehydrogenase/reductase family, cf. EC 1.1.1.51
additional information
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beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase
additional information
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enzyme complex 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase
additional information
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DELTA5-3-beta-hydroxysteroid dehydrogenase-isomerase system
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a 3-oxo-DELTA5-steroid = a 3-oxo-DELTA4-steroid
mechanism in which the transition state for enolization is dienolate-like, characterized by relatively little proton transfer from Tyr14 in the transition state, and the intermediate in the overall reaction is dienol-like. An alternative mechanism in which the intermediate is stabilized by a short, strong hydrogen bond can also be consistent with the data
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a 3-oxo-DELTA5-steroid = a 3-oxo-DELTA4-steroid
"flip-flop" mechanism may be involved
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a 3-oxo-DELTA5-steroid = a 3-oxo-DELTA4-steroid
catalytic mechanism, phenolate binding to the oxyanion hole of ketosteroid isomerase via hydrogen bonding, electrostatic contributions and geometric and electrostatic changes, overview, the KSI reaction involves changes in both geometry and charge distribution as the reaction proceeds from its ground state to its intermediate
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a 3-oxo-DELTA5-steroid = a 3-oxo-DELTA4-steroid
catalytic mechanism, phenolate binding to the oxyanion hole of ketosteroid isomerase via hydrogen bonding, electrostatic contributions and geometric and electrostatic changes, overview, the KSI reaction involves changes in both geometry and charge distribution as the reaction proceeds from its ground state to its intermediate, Tyr16 and Asp103 are important in catalysis
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a 3-oxo-DELTA5-steroid = a 3-oxo-DELTA4-steroid
two-step reaction first, Asp38 acts as a base, abstracting the 4beta-H atom from C-4 of the steroid to form a dienolate as the intermediate, next, the intermediate is reketonized by proton transfer to the 6beta-position, each step goes through its own transition state, catalytic mechanism, theoretical model involving residues Tyr14, Asp38 and Asp99
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a 3-oxo-DELTA5-steroid = a 3-oxo-DELTA4-steroid
enzyme enhances the coupled motion/hydrogen tunneling contribution to the rate acceleration over the solution reaction
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a 3-oxo-DELTA5-steroid = a 3-oxo-DELTA4-steroid
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
intramolecular oxidoreduction
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isomerization
isomerization
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isomerization
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catalysis of the allylic isomerization of a variety of DELTA5-3-ketosteroids by intramolecular transfer of the C4-beta proton to the C6-beta position
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PATHWAY SOURCE
PATHWAYS
KEGG
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, androgen and estrogen metabolism, androgen and estrogen metabolism, androgen and estrogen metabolism, androgen and estrogen metabolism, androgen and estrogen metabolism, androgen and estrogen metabolism, androgen and estrogen metabolism, Microbial metabolism in diverse environments, Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
3-oxosteroid DELTA5-DELTA4-isomerase
This activity is catalysed by several distinct enzymes (cf. EC 1.1.3.6, cholesterol oxidase and EC 1.1.1.145, 3-hydroxy-5-steroid dehydrogenase).
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CAS REGISTRY NUMBER
COMMENTARY
9031-36-1
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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
(+)-cis-isopulegone
(+)-pulegone
the enzyme also exhibits (+)-cis-isopulegone isomerase activity
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?
androst-5-ene-3,17-dione
androst-4-ene-3,17-dione
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alpha-secondary deuterium kinetic isotope effects at C6 of the steroid. Presence of coupled motion/hydrogen tunneling in the enzyme-catalyzed reaction
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r
17beta-Hydroxy-5(10)-estren-3-one
?
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at 0.25% of the activity compared to 5-androstene-3,17-dione
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?
5(10)-estrene-3,17-dione
estr-4-en-3,17-dione
nonsticky substrate
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?
5-Androstene-3,17-dione
4-Androstene-3,17-dione
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?
5-Androstene-3,17-dione
4-Androstene-3,17-dione
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?
5-Androstene-3,17-dione
4-Androstene-3,17-dione
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residue Asp38 acts as a base to abstract a proton from C-4 of the substrate to form an intermediate dienolate, which is then reprotonated on C-6
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?
5-Androstene-3,17-dione
4-Androstene-3,17-dione
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?
5-Androstene-3,17-dione
4-Androstene-3,17-dione
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the hydrogen-bond network which links the two catalytic residues, Tyr14 and Asp99, to Tyr30, Tyr55, and a water molecule in the highly apolar active site provides the structural support that is needed for the enzyme to maintain the active site geometry optimized for both function and stability
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?
5-Pregnene-3,20-dione
4-Pregnene-3,20-dione
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?
a 3-oxo-DELTA5-steroid
a 3-oxo-DELTA4-steroid
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phenolates binding to the oxyanion hole of the enzyme via electrostatic interactions, different binding of transition state analogue and substrate, hydrogen bonds shorten with increasing charge localization, overview
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r
a 3-oxo-DELTA5-steroid
a 3-oxo-DELTA4-steroid
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phenolates binding to the oxyanion hole of the enzyme via hydrogen bonding, different binding of transition state analogue and substrate, hydrogen bonds shorten with increasing charge localization, overview
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r
additional information
?
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identification of a putative 3beta-hydroxy-steroid dehydrogenase/DELTA5-DELTA4 isomerase pathway with the ability to handle both dehydroepiansdrosterone as well as C-17 side-chain containing compounds such as pregnenolone and 3beta-hydroxy-16alpha,17alpha-epoxypregn-5-en-20-one. 3beta-HSD/isomerase activity only occurs following lactonization of the steroidal ring-D. Presence of C-7 allylic hydroxylation, in either epimeric form, inhibits 3beta-HSD/isomerase activity. In Aspergillus tamarii, pregnenolone and 3beta-hydroxy-16alpha,17alpha-epoxypregn-5-en-20-one are metabolized solely through the putative 3beta-HSD/isomerase pathway
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?
additional information
?
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identification of a putative 3beta-hydroxy-steroid dehydrogenase/DELTA5-DELTA4 isomerase pathway with the ability to handle both dehydroepiansdrosterone as well as C-17 side-chain containing compounds such as pregnenolone and 3beta-hydroxy-16alpha,17alpha-epoxypregn-5-en-20-one. 3beta-HSD/isomerase activity only occurs following lactonization of the steroidal ring-D. Presence of C-7 allylic hydroxylation, in either epimeric form, inhibits 3beta-HSD/isomerase activity. In Aspergillus tamarii, pregnenolone and 3beta-hydroxy-16alpha,17alpha-epoxypregn-5-en-20-one are metabolized solely through the putative 3beta-HSD/isomerase pathway
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?
additional information
?
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properties of the steroid-enzyme aduct
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?
additional information
?
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the 2.3 A structure indicates that Phe101 is not a catalytic residue and that Tyr14 and Asp99 COOH should be directly involved in the stabilization of the dienolate intermediate
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?
additional information
?
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the 2.3 A structure indicates that Phe101 is not a catalytic residue and that Tyr14 and Asp99 COOH should be directly involved in the stabilization of the dienolate intermediate
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?
additional information
?
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equilenin geometrically and electrostatically resembles the dienolate reaction intermediate and transition state, binding structure to enzyme mutant D40N, overview
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?
additional information
?
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mechanism of KSI-catalyzed steroid isomerization, overview
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?
additional information
?
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the reaction occurs by a two-step general acid-base mechanism involving a dienolate intermediate. Hybrid quantum/classical molecular dynamics simulations elucidating the geometrical, conformational, and electrostatic changes occurring during the isomerization reaction catalyzed by KSI, calculation of rate constants and modeling of the KSI active site, overview. Electrostaic and conformational changes during the proton transfer reactions, rearrangements, overview
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?
additional information
?
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in pregnancy placental 3beta-hydroxy-5-ene-steroid dehydrogenase and steroid 5--4 isomerase produce progesterone from pregnenolone and metabolize fetal dehydroepiandrosterone sulfate to androstenedione, an estrogen precursor
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?
additional information
?
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the conversion of 3beta-hydroxy-5-ene steroids by the enzyme complex 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase is an obligatory step in the biosynthesis of all classes of hormonal steroids in classical steroidogenic as well as in peripheral tissues
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?
additional information
?
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the bifunctional enzyme possesses 3beta-hydroxysteroid dehydrogenase activity, EC 1.1.1.51, and steroid DELTA-isomerase activity, the enzyme is a key steroidogenic enzyme that catalyzes the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
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?
additional information
?
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the bifunctional enzyme possesses 3beta-hydroxysteroid dehydrogenase activity, EC 1.1.1.51, and steroid DELTA-isomerase activity, the enzyme is a key steroidogenic enzyme that catalyzes the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
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?
additional information
?
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the bifunctional enzyme possesses 3beta-hydroxysteroid dehydrogenase activity, EC 1.1.1.51, and steroid DELTA-isomerase activity, the enzyme is a key steroidogenic enzyme that catalyzes the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
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?
additional information
?
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structure-function relations, analysis of three dimensional model of a ternary complex of human 3beta-HSD type 1 with an NAD+ cofactor and androstenedione product, reaction of EC 1.1.1.51, to elucidate the key substrate binding residues in the active site as well as the basis for dual function of the 3beta-HSD_1 enzyme, Asn100 and Glu126 residues are key residues that participate in the dehydrogenase and isomerization reactions, respectively, isomerase substrate binding structure, overview
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?
additional information
?
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structure-function relations, analysis of three dimensional model of a ternary complex of human 3beta-HSD type 1 with an NAD+ cofactor and androstenedione product, reaction of EC 1.1.1.51, to elucidate the key substrate binding residues in the active site as well as the basis for dual function of the 3beta-HSD_1 enzyme, Asn100 and Glu126 residues are key residues that participate in the dehydrogenase and isomerization reactions, respectively, isomerase substrate binding structure, overview
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?
additional information
?
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structure-function relations, analysis of three dimensional model of a ternary complex of human 3beta-HSD type 1 with an NAD+ cofactor and androstenedione product, reaction of EC 1.1.1.51, to elucidate the key substrate binding residues in the active site as well as the basis for dual function of the 3beta-HSD_1 enzyme, Asn100 and Glu126 residues are key residues that participate in the dehydrogenase and isomerization reactions, respectively, isomerase substrate binding structure, overview
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?
additional information
?
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human GST A3-3 does not stabilize a dienolate by an oxyanion hole in the active site
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?
additional information
?
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human GST A3-3 does not stabilize a dienolate by an oxyanion hole in the active site
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?
additional information
?
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the conversion of 3beta-hydroxy-5-ene steroids by the enzyme complex 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase is an obligatory step in the biosynthesis of all classes of hormonal steroids in classical steroidogenic as well as in peripheral tissues
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?
additional information
?
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the aromatic residues Phe54, Phe82 and Trp116 in the hydrophobic substrate-binding pocket of the enzyme contribute differentially to steroid species including substrate, intermediate and product
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?
additional information
?
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enzyme can be induced by various steroids
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?
additional information
?
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mechanism of KSI-catalyzed steroid isomerization, direct detection of physical coupling between the Y16 and D103 hydrogen bonds, ligand binding, overview
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?
additional information
?
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the enzyme catalyzes a C-H bond cleavage and formation through an enolate intermediate. Conversion of the ketone substrate to the enolate intermediate is simulated by a photoacid bound to the active site oxyanion hole. The lack of a significant solvation response in KSI suggests a preorganized active site
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?
additional information
?
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bifunctional enzyme possessing 3-beta-hydroxy-DELTA5-steroid dehydrogenase, EC 1.1.1.145, and steroid DELTA-isomerase activity
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?
additional information
?
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the enzyme is involved in synthesis of sex steroids influencing the development and function of the songbird brain, overview
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?
additional information
?
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bifunctional enzyme possessing 3beta-hydroxysteroid dehydrogenase activity, EC 1.1.1.51, and steroid DELTA-isomerase activity
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?
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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
5-Androstene-3,17-dione
4-Androstene-3,17-dione
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-
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?
additional information
?
-
-
in pregnancy placental 3beta-hydroxy-5-ene-steroid dehydrogenase and steroid 5--4 isomerase produce progesterone from pregnenolone and metabolize fetal dehydroepiandrosterone sulfate to androstenedione, an estrogen precursor
-
-
?
additional information
?
-
-
the conversion of 3beta-hydroxy-5-ene steroids by the enzyme complex 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase is an obligatory step in the biosynthesis of all classes of hormonal steroids in classical steroidogenic as well as in peripheral tissues
-
-
?
additional information
?
-
the bifunctional enzyme possesses 3beta-hydroxysteroid dehydrogenase activity, EC 1.1.1.51, and steroid DELTA-isomerase activity, the enzyme is a key steroidogenic enzyme that catalyzes the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
-
-
?
additional information
?
-
the bifunctional enzyme possesses 3beta-hydroxysteroid dehydrogenase activity, EC 1.1.1.51, and steroid DELTA-isomerase activity, the enzyme is a key steroidogenic enzyme that catalyzes the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
-
-
?
additional information
?
-
-
the bifunctional enzyme possesses 3beta-hydroxysteroid dehydrogenase activity, EC 1.1.1.51, and steroid DELTA-isomerase activity, the enzyme is a key steroidogenic enzyme that catalyzes the first step of the multienzyme pathway conversion of circulating dehydroepiandrosterone and pregnenolone to active steroid hormones
-
-
?
additional information
?
-
-
the conversion of 3beta-hydroxy-5-ene steroids by the enzyme complex 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase is an obligatory step in the biosynthesis of all classes of hormonal steroids in classical steroidogenic as well as in peripheral tissues
-
-
?
additional information
?
-
-
enzyme can be induced by various steroids
-
-
?
additional information
?
-
-
the enzyme is involved in synthesis of sex steroids influencing the development and function of the songbird brain, overview
-
-
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADPH
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D36A/K37R mutation shifts the cofactor preference of both 3-beta-hydroxysteroid dehydrogenase and isomerase from NAD(H) to NADP(H)
NADH
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the isomerase activity of the wild-type enzyme and the deletion mutant enzyme DELTA283-310 requires allosteric activation by NADH, Km for wild-type enzyme and deletion mutant enzyme is 0.0046 mM
NADH
-
wild-type 1 isomerase is activated only by NADH with NADPH producing no isomerase activity. D36A/K37R mutation shifts the cofactor preference of both 3-beta-hydroxysteroid dehydrogenase and isomerase from NAD(H) to NADP(H)
NADH
-
isomerase activity of enzyme requires NADH, Km- and kcat-value of isoform 1 wild-type are 0.0046 mM and 45 per min, resp. Km- and kcat-value of isoform 2 wild-type are 0.0126 mM and 99.1 per min, resp
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(17S)-Spiro[5alpha-androstane-17,2'-oxiran]-3-one
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potent irreversible active-site directed
(17S)-Spiro[estra-1,3,5(10),6,8-pentaene-17,2'-oxiran]-3-ol
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active-site-directed, irreversible
(17S)-Spiro[estra-1,3,5(10)-triene-17,2'-oxiran]-3-ol
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potent irreversible active-site directed
10beta-(1-Oxoprop-2-ynyl)oestr-4-ene-3,17-dione
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irreversible, time-dependent, active-site directed
17beta-(1-Oxoprop-2-ynyl)androst-4-en-3-one
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irreversible, time-dependent, active-site directed
17beta-Hydroxy-5,10-seco-oestr-4-yne-3,10-dione
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irreversible, time-dependent, active-site directed
5,10-Seco-oestr-4-yne-3,10,17-trione
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irreversible, time-dependent, active-site directed
coumarin 183
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binds tightly in the oxyanion hole of the enzyme and chemically resemble the intermediate in the catalytic cycle. Upon photoexcitation, the pKa of the hydroxyl group changes substantially. This light-activated change in electron density around the photoacid hydroxyl group simulates the change in substrate pKa during the catalytic cycle. When the light-driven reaction analog occurs in the KSI active site, the electrostatic environment changes little
Halothane
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allosteric modulation of dynamics-function relationship without direct competition, halothane occupancy at the dimer interface disrupts intersubunit hydrogen bonding
N,N-Dimethyl-4-methyl-3-oxo-4-aza-5alpha-androstane-17beta-carboxamide
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non-competitive inhibition of isomerase, competitive inhibition of 3beta-hydroxysteroid dehydrogenase
Spiro-17beta-oxiranyl-DELTA4-androsten-3-one(4beta)
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active site-directed, irreversible
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Spiro-17beta-oxiranylestra-1,3,5(10),6,8-pentaene-3-ol
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active site-directed, irreversible
equilenin
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reaction intermediate analogue, binds to the active site, binding structure, overview
equilenin
reaction intermediate analogue, binds to the active site, binding structure, overview
equilenin
-
binds tightly in the oxyanion hole of the enzyme and chemically resemble the intermediate in the catalytic cycle. Upon photoexcitation, the pKa of the hydroxyl group changes substantially. This light-activated change in electron density around the photoacid hydroxyl group simulates the change in substrate pKa during the catalytic cycle. When the light-driven reaction analog occurs in the KSI active site, the electrostatic environment changes little
Urea
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enzymatic activity is affected significantly by urea and decreases exponentially as urea concentration increases
additional information
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overview: interaction of the enzyme with steroids
-
additional information
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overview: inactivation by active-site directed acetylenic steroids
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additional information
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inhibitor development, a series of substituted naphthol inhibitors shows substantial variations in the contributions of the protonated and deprotonated forms when bound to the active site. Method of quantifying the contributions of each protonation state show the oxyanion hole in the active site of wild-type DELTA5-3-ketosteroid isomerase to have a proton affinity equal to a solution pKa of 10.05, which is similar to the measured pKa of 10.0 of the reaction intermediate
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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DISEASE
TITLE OF PUBLICATION
LINK TO PUBMED
3beta-hydroxy-delta5-steroid dehydrogenase deficiency
Carboxyl-Terminal Mutations in 3{beta}-Hydroxysteroid Dehydrogenase Type II Cause Severe Salt-Wasting Congenital Adrenal Hyperplasia.
Addison Disease
Steroid-cell autoantibodies are preferentially expressed in women with premature ovarian failure who have adrenal autoimmunity.
Adenoma
Immunohistochemical study of cytochrome b5 in human adrenal gland and in adrenocortical adenomas from patients with Cushing's syndrome.
Adrenal Hyperplasia, Congenital
A new insight into the molecular basis of 3beta-hydroxysteroid dehydrogenase deficiency.
Adrenal Hyperplasia, Congenital
A novel nonstop mutation in the stop codon and a novel missense mutation in the type II 3beta-hydroxysteroid dehydrogenase (3beta-HSD) gene causing, respectively, nonclassic and classic 3beta-HSD deficiency congenital adrenal hyperplasia.
Adrenal Hyperplasia, Congenital
Congenital adrenal hyperplasia due to 3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4) isomerase deficiency.
Adrenal Hyperplasia, Congenital
Molecular biology of the 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase gene family.
Adrenal Hyperplasia, Congenital
New insight into the molecular basis of 3beta-hydroxysteroid dehydrogenase deficiency: identification of eight mutations in the HSD3B2 gene eleven patients from seven new families and comparison of the functional properties of twenty-five mutant enzymes.
Adrenal Hyperplasia, Congenital
Newly proposed hormonal criteria via genotypic proof for type II 3beta-hydroxysteroid dehydrogenase deficiency.
Adrenocortical Hyperfunction
Trilostane treatment in dogs with pituitary-dependent hyperadrenocorticism.
Anovulation
Prepubertal administration of estradiol valerate disrupts cyclicity and leads to cystic ovarian morphology during adult life in the rat: role of sympathetic innervation.
Autoimmune Diseases
Steroid-cell autoantibodies are preferentially expressed in women with premature ovarian failure who have adrenal autoimmunity.
Brain Injuries, Traumatic
3beta-Hydroxysteroid dehydrogenase/5-ene-4-ene isomerase mRNA expression in rat brain: effect of pseudopregnancy and traumatic brain injury.
Breast Neoplasms
Crucial role of cytokines in sex steroid formation in normal and tumoral tissues.
Breast Neoplasms
Identification of key amino acids responsible for the substantially higher affinities of human type 1 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD1) for substrates, coenzymes, and inhibitors relative to human 3beta-HSD2.
Breast Neoplasms
Induction of 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase type 1 gene transcription in human breast cancer cell lines and in normal mammary epithelial cells by interleukin-4 and interleukin-13.
Breast Neoplasms
Induction of 3beta-hydroxysteroid dehydrogenase/isomerase type 1 expression by interleukin-4 in human normal prostate epithelial cells, immortalized keratinocytes, colon, and cervix cancer cell lines.
Breast Neoplasms
Multiple signal transduction pathways mediate interleukin-4-induced 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4 isomerase in normal and tumoral target tissues.
Breast Neoplasms
Multiple signaling pathways mediate interleukin-4-induced 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase type 1 gene expression in human breast cancer cells.
Breast Neoplasms
Regulation of sex steroid formation by interleukin-4 and interleukin-6 in breast cancer cells.
Breast Neoplasms
Serine 124 completes the Tyr, Lys and Ser triad responsible for the catalysis of human type 1 3beta-hydroxysteroid dehydrogenase.
Breast Neoplasms
Structural basis for the selective inhibition of human 3beta-hydroxysteroid dehydrogenase 1 in human breast tumor MCF-7 cells.
Carcinoma
Ontogeny and subcellular localization of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) in the human and rat adrenal, ovary and testis.
Carcinoma
StAR and progesterone producing enzymes (3beta-hydroxysteroid dehydrogenase and cholesterol side-chain cleavage cytochromes P450) in human epithelial ovarian carcinoma: immunohistochemical and real-time PCR studies.
Carcinoma
Structure and tissue-specific expression of 3 beta-hydroxysteroid dehydrogenase/5-ene-4-ene isomerase genes in human and rat classical and peripheral steroidogenic tissues.
Carcinoma
Structure, function and tissue-specific gene expression of 3?-hydroxysteroid dehydrogenase/5-ene-4-ene isomerase enzymes in classical and peripheral intracrine steroidogenic tissues.
Carcinoma
Synergistic activation of the human type II 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase promoter by the transcription factor steroidogenic factor-1/adrenal 4-binding protein and phorbol ester.
Carcinoma, Hepatocellular
Hepatocellular ligandin during N-2-fluorenylacetamide carcinogenesis.
Carcinoma, Hepatocellular
Human cytosolic 3alpha-hydroxysteroid dehydrogenases of the aldo-keto reductase superfamily display significant 3beta-hydroxysteroid dehydrogenase activity: implications for steroid hormone metabolism and action.
Cholera
Regulation of steroidogenesis in jc-410, a stable cell line of porcine granulosa origin.
Cholera
Steroid regulation of progesterone synthesis in a stable porcine granulosa cell line: a role for progestins.
Cholera
Time-course effects of human recombinant luteinizing hormone on porcine Leydig cell specific differentiated functions.
Cholestasis
3beta-hydroxy-delta5 -C27-steroid dehydrogenase deficiency: diagnosis and treatment.
Cholestasis, Intrahepatic
The bile acid synthetic gene 3beta-hydroxy-Delta(5)-C(27)-steroid oxidoreductase is mutated in progressive intrahepatic cholestasis.
Choriocarcinoma
Induction of 3beta-hydroxysteroid dehydrogenase/isomerase type 1 expression by interleukin-4 in human normal prostate epithelial cells, immortalized keratinocytes, colon, and cervix cancer cell lines.
Choriocarcinoma
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.
Colonic Neoplasms
Induction of 3beta-hydroxysteroid dehydrogenase/isomerase type 1 expression by interleukin-4 in human normal prostate epithelial cells, immortalized keratinocytes, colon, and cervix cancer cell lines.
Cysts
Dexamethasone-induced changes in sympathetic innervation of porcine ovaries and in their steroidogenic activity.
Cysts
Immunolocalization of 3beta-HSD and 17beta-HSD in the testis of the spotted ray Torpedo marmorata.
Demyelinating Diseases
Distribution of 3beta-hydroxysteroid dehydrogenase in the cerebellum in canine distemper virus infection.
Diabetes Mellitus
Steroid-cell autoantibodies are preferentially expressed in women with premature ovarian failure who have adrenal autoimmunity.
Diabetes Mellitus, Type 1
Steroid-cell autoantibodies are preferentially expressed in women with premature ovarian failure who have adrenal autoimmunity.
Disorders of Sex Development
Newly proposed hormonal criteria via genotypic proof for type II 3beta-hydroxysteroid dehydrogenase deficiency.
Fetal Resorption
Effect of abdominal vagotomy of the pregnant rat on LH and progesterone concentrations and fetal resorption.
Genetic Diseases, Inborn
[Case report: congenital adrenal hyperplasia and ambiguous genitalia due to 3 beta-hydroxysteroid dehydrogenase deficiency]
Herpes Zoster
Ontogeny of 3beta-hydroxysteroid dehydrogenase expression in the rat adrenal gland as studied by immunohistochemistry.
Herpes Zoster
The primate adrenal zona reticularis is defined by expression of cytochrome b5, 17alpha-hydroxylase/17,20-lyase cytochrome P450 (P450c17) and NADPH-cytochrome P450 reductase (reductase) but not 3beta-hydroxysteroid dehydrogenase/delta5-4 isomerase (3beta-HSD).
Hirsutism
The Molecular and Clinical Spectrum of 3beta-hydroxysteroid Dehydrogenase Deficiency Disorder.
Infections
Characterization of the Rana grylio virus 3beta-hydroxysteroid dehydrogenase and its novel role in suppressing virus-induced cytopathic effect.
Infections
Distribution of 3beta-hydroxysteroid dehydrogenase in the cerebellum in canine distemper virus infection.
Infections
HIV-1 clade C infection and progressive disruption in the relationship between cortisol, DHEAS and CD4 cell numbers: a two-year follow-up study.
Neoplasms
Cushing's syndrome due to bilateral adrenocortical adenomas with different pathological features.
Neoplasms
Immunohistochemical detection of inhibin-alpha, -betaB, and -betaA chains and 3beta-hydroxysteroid dehydrogenase in canine testicular tumors and normal testes.
Neoplasms
Multiple signal transduction pathways mediate interleukin-4-induced 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4 isomerase in normal and tumoral target tissues.
Neoplasms
Multiple signaling pathways mediate interleukin-4-induced 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase type 1 gene expression in human breast cancer cells.
Neoplasms
StAR and progesterone producing enzymes (3beta-hydroxysteroid dehydrogenase and cholesterol side-chain cleavage cytochromes P450) in human epithelial ovarian carcinoma: immunohistochemical and real-time PCR studies.
Neoplasms
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.
Neoplasms
Use of antibodies against LH receptor, 3beta-hydroxysteroid dehydrogenase and vimentin to characterize different types of testicular tumour in dogs.
Optic Atrophy, Autosomal Dominant
Protease Inhibitor Anti-HIV, Lopinavir, Impairs Placental Endocrine Function.
Ovarian Cysts
Prepubertal administration of estradiol valerate disrupts cyclicity and leads to cystic ovarian morphology during adult life in the rat: role of sympathetic innervation.
Peripheral Nerve Injuries
Differential regulation of 3-beta-hydroxysteroid dehydrogenase and vanilloid receptor TRPV1 mRNA in sensory neurons by capsaicin and NGF.
Peripheral Nerve Injuries
Increased expression of 3 beta-hydroxysteroid dehydrogenase mRNA in dorsal root ganglion neurons of adult rats following peripheral nerve injury.
Pregnancy, Ectopic
Expression of 3beta-hydroxysteroid dehydrogenase and P450 side chain cleavage enzyme in the human uterine endometrium.
Primary Ovarian Insufficiency
Steroid-cell autoantibodies are preferentially expressed in women with premature ovarian failure who have adrenal autoimmunity.
Prostatic Neoplasms
Induction of 3beta-hydroxysteroid dehydrogenase/isomerase type 1 expression by interleukin-4 in human normal prostate epithelial cells, immortalized keratinocytes, colon, and cervix cancer cell lines.
Seminoma
Immunohistochemical detection of inhibin-alpha, -betaB, and -betaA chains and 3beta-hydroxysteroid dehydrogenase in canine testicular tumors and normal testes.
Seminoma
Use of antibodies against LH receptor, 3beta-hydroxysteroid dehydrogenase and vimentin to characterize different types of testicular tumour in dogs.
steroid delta-isomerase deficiency
3beta-hydroxy-delta5 -C27-steroid dehydrogenase deficiency: diagnosis and treatment.
steroid delta-isomerase deficiency
A new insight into the molecular basis of 3beta-hydroxysteroid dehydrogenase deficiency.
steroid delta-isomerase deficiency
A novel homozygous nonsense mutations E135* in the type II 3beta-hydroxysteroid dehydrogenase gene in a girl with salt-losing congenital adrenal hyperplasia. Mutations in brief no. 168. Online.
steroid delta-isomerase deficiency
A novel nonstop mutation in the stop codon and a novel missense mutation in the type II 3beta-hydroxysteroid dehydrogenase (3beta-HSD) gene causing, respectively, nonclassic and classic 3beta-HSD deficiency congenital adrenal hyperplasia.
steroid delta-isomerase deficiency
An adrenal rest tumour in the perirenal region in a patient with congenital adrenal hyperplasia due to congenital 3beta-hydroxysteroid dehydrogenase deficiency.
steroid delta-isomerase deficiency
Carboxyl-Terminal Mutations in 3{beta}-Hydroxysteroid Dehydrogenase Type II Cause Severe Salt-Wasting Congenital Adrenal Hyperplasia.
steroid delta-isomerase deficiency
Congenital adrenal hyperplasia due to 3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4) isomerase deficiency.
steroid delta-isomerase deficiency
Delayed diagnosis of congenital adrenal hyperplasia with salt wasting due to type II 3beta-hydroxysteroid dehydrogenase deficiency.
steroid delta-isomerase deficiency
Endocrinological and genetic studies in patients with Polycystic Ovary Syndrome (PCOS).
steroid delta-isomerase deficiency
Molecular biology of the 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase gene family.
steroid delta-isomerase deficiency
Molecular genetics of 3beta-hydroxy-Delta5-C27-steroid oxidoreductase deficiency in 16 patients with loss of bile acid synthesis and liver disease.
steroid delta-isomerase deficiency
Newly proposed hormonal criteria via genotypic proof for type II 3beta-hydroxysteroid dehydrogenase deficiency.
steroid delta-isomerase deficiency
Oral cholic acid for hereditary defects of primary bile acid synthesis: a safe and effective long-term therapy.
steroid delta-isomerase deficiency
The Molecular and Clinical Spectrum of 3beta-hydroxysteroid Dehydrogenase Deficiency Disorder.
steroid delta-isomerase deficiency
[Case report: congenital adrenal hyperplasia and ambiguous genitalia due to 3 beta-hydroxysteroid dehydrogenase deficiency]
Testicular Neoplasms
Immunohistochemical detection of inhibin-alpha, -betaB, and -betaA chains and 3beta-hydroxysteroid dehydrogenase in canine testicular tumors and normal testes.
Thiamine Deficiency
Effects of thiamine deficiency on hepatic cytochromes P450 and drug-metabolizing enzyme activities.
Uterine Cervical Neoplasms
Induction of 3beta-hydroxysteroid dehydrogenase/isomerase type 1 expression by interleukin-4 in human normal prostate epithelial cells, immortalized keratinocytes, colon, and cervix cancer cell lines.
Vaccinia
Steroid hormone synthesis by vaccinia virus suppresses the inflammatory response to infection.
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0171
5-androstene-3,17-dione
mutant enzyme Y14F, at pH 7.0, temperature not specified in the publication
0.023
5-androstene-3,17-dione
mutant enzyme Y55F, at pH 7.0, temperature not specified in the publication
0.0252
5-androstene-3,17-dione
-
22°C, pH 7.4, truncated cytosolic enzyme DEALTA283-310
0.0417
5-androstene-3,17-dione
mutant enzyme Y115F, at pH 7.0, temperature not specified in the publication
0.0486
5-androstene-3,17-dione
25°C, pH 7.0, mutant enzyme F116W
0.0499
5-androstene-3,17-dione
wild type enzyme, at pH 7.0, temperature not specified in the publication
0.0502
5-androstene-3,17-dione
mutant enzyme Y30F/Y55F, at pH 7.0, temperature not specified in the publication
0.0513
5-androstene-3,17-dione
-
25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F
0.0552
5-androstene-3,17-dione
mutant enzyme Y30F, at pH 7.0, temperature not specified in the publication
0.0578
5-androstene-3,17-dione
-
25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F/D99N
0.067
5-androstene-3,17-dione
-
25°C, pH 6.5, solvent D2O, mutant enzyme D38A
0.073
5-androstene-3,17-dione
-
25°C, pH 8.5, solvent D2O, mutant enzyme D38A
0.0773
5-androstene-3,17-dione
mutant enzyme Y30F/Y55F/Y115F, at pH 7.0, temperature not specified in the publication
0.085
5-androstene-3,17-dione
-
25°C, pH 7.5, solvent D2O, mutant enzyme D38A
0.0918
5-androstene-3,17-dione
-
25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F/D99L
0.094
5-androstene-3,17-dione
-
25°C, pH 8.5, solvent H2O, mutant enzyme D38A
0.096
5-androstene-3,17-dione
-
25°C, pH 7.5, solvent H2O, mutant enzyme D38A
0.099
5-androstene-3,17-dione
-
25°C, pH 6.5, solvent H2O, mutant enzyme D38A
additional information
additional information
kinetics of wild-type and mutant enzymes
-
additional information
additional information
kinetics of wild-type and mutant enzymes
-
additional information
additional information
-
kinetics of wild-type and mutant enzymes
-
additional information
additional information
-
Km-values of mutant enzymes Y16F and D40N
-
additional information
additional information
thermodynamic parameters and rate constants for isomerization of 5-androstene-3,17-dione
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
turnover number of mutant enzymes Y16F and D40N
-
0.052
5-androstene-3,17-dione
-
25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F/D99L
1.1
5-androstene-3,17-dione
-
25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F/D99N
13.3
5-androstene-3,17-dione
mutant enzyme Y14F, at pH 7.0, temperature not specified in the publication
230
5-androstene-3,17-dione
-
25°C, pH 6.5, solvent D2O, mutant enzyme D38A
260
5-androstene-3,17-dione
-
25°C, pH 8.5, solvent D2O, mutant enzyme D38A
279
5-androstene-3,17-dione
-
25°C, pH 6.5, solvent H2O, mutant enzyme D38A
340
5-androstene-3,17-dione
-
25°C, pH 6.5, solvent H2O, mutant enzyme D38A
412
5-androstene-3,17-dione
-
25°C, pH 7.5 or 8.5, solvent H2O, mutant enzyme D38A
3510
5-androstene-3,17-dione
mutant enzyme Y55F, at pH 7.0, temperature not specified in the publication
10700
5-androstene-3,17-dione
mutant enzyme Y30F/Y55F, at pH 7.0, temperature not specified in the publication
12200
5-androstene-3,17-dione
mutant enzyme Y30F/Y55F/Y115F, at pH 7.0, temperature not specified in the publication
14500
5-androstene-3,17-dione
mutant enzyme Y115F, at pH 7.0, temperature not specified in the publication
17800
5-androstene-3,17-dione
mutant enzyme Y30F, at pH 7.0, temperature not specified in the publication
21000
5-androstene-3,17-dione
wild type enzyme, at pH 7.0, temperature not specified in the publication
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
780
5-androstene-3,17-dione
mutant enzyme Y14F, at pH 7.0, temperature not specified in the publication
150000
5-androstene-3,17-dione
mutant enzyme Y55F, at pH 7.0, temperature not specified in the publication
160000
5-androstene-3,17-dione
mutant enzyme Y30F/Y55F/Y115F, at pH 7.0, temperature not specified in the publication
210000
5-androstene-3,17-dione
mutant enzyme Y30F/Y55F, at pH 7.0, temperature not specified in the publication
330000
5-androstene-3,17-dione
mutant enzyme Y30F, at pH 7.0, temperature not specified in the publication
350000
5-androstene-3,17-dione
mutant enzyme Y115F, at pH 7.0, temperature not specified in the publication
430000
5-androstene-3,17-dione
wild type enzyme, at pH 7.0, temperature not specified in the publication
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
Ki-values for equilenin and 19-nortestosterone, wild-type enzyme and mutant enzyme D99E
-
additional information
additional information
-
dissociation constants of inhibitors
-
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.622
-
reaction with 5-androsten-3,17-dione, truncated cytosolic enzyme DELTA283-310
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 8
7.5 - 9
-
pH 7.5: about 40% of maximal activity, pH 9.0: about 60% of maximal activity
6.5 - 8
-
pH 6.5: about 60% of maximal activity, pH 8.0: about 40% of maximal activity, pH 6.3: no activity
6.5 - 8
-
pH 6.5: about 55% of maximal activity, wild-type enzyme and mutant enzyme D265N, about 65% of maximal activity, mutant enzyme D241N, about 85% of maximal activity mutant enzyme D257L and D258L. pH 8.0: about 50% of maximal activity, wild-type enzyme and mutant enzymes D241N, 95% of maximal activity, mutant enzyme D258L
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
22 - 42
-
22°C: about 60% of maximal activity, 40°C: about 40% of maximal activity
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
3beta-hydroxy-5-ene steroid dehydrogenase/steroid DELTA-isomerase enzyme system
-
-
brenda
3beta-hydroxy-5-ene-steroid dehydrogenase/steroid DELTA5-DELTA4-ene isomerase enzyme complex
-
-
brenda
-
-
-
brenda
bifunctional 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4-isomerase, isoform 3betaHSD1
-
-
brenda
bifunctional 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4-isomerase, isoform 3betaHSD2
-
-
brenda
bifunctional 3beta-hydroxysteroid dehydrogenase/isomerase, isoforms type 1 and 2
-
-
brenda
enzyme complex of 3beta-hydroxy-5-ene-steroid dehydrogenase and Steroid DELTA-isomerase
-
-
brenda
biotype B
-
-
brenda
type I enzyme isoform, bifunctional 3beta-hydroxysteroid dehydrogenase/D5-D4 isomerase
UniProt
brenda
bifunctional 3beta-hydroxysteroid dehydrogenase/D5-D4 isomerase
-
-
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
of embryo. Levels of bifunctional 3beta-hydroxysteroid-dehydrogenase/DELTA4-DELTA5-isomerase are sexually dimorphic and developmental age-dependent. MRNA levels are higher in females than in males at E7, whereas this dimorphism is reversed at E9 and E15. In females, the highest mRNA level is at E7, whereas in males the levels are significantly higher at E9 and E17 than at E7 and E11. Sexual dimorphism can only be detcted in prosencephalon, can not be observed before gonadal sexual differentiation, and is not paralleled by a dimorphism in brain content of progesterone
brenda
enzyme expression analysis in male and pseudopregnant female rat brain, in situ hybridization, overview, strong expression in olfactory bulb, striatum, cortex, thalamus, hypothalamus, septum, habenula, hippocampus and cerebellum, pseudopregnancy has no effect on enzyme expression in brain, effect of lesions, overview
brenda
-
brain region differences in dehydroepiandrosterone metabolism by enzyme, sex differences in enzyme distribution and regulation by stress
brenda
in ovariectomized-adrenalectomized animals, estrogen treatment increases enzyme mRNA in the hypothalamus as well as enzyme activity
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
modified enzyme in which the membrane-spanning domain, residues 283-310, of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells
brenda
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
-
the enzyme is responsible for progesterone and androstenedione production
Show AA Sequence and Transmembrane Helices (410 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
11000
-
2 * 11000, SDS-PAGE. At concentrations below 1 mg/ml the enzyme exists as a dimer of MW 26800, determined by gel filtration and ultracentrifugation studies. Concentration-dependent association occurs at higher enzyme concentrations
14000
-
2 * 14000, mutant enzymnes R72A, E118A, N120A and E18A/N120A, SDS-PAGE
14520
19000
-
4 * 19000, enzyme complex of 3beta-hydroxy-5-ene-steroid dehydrogenase and steroid DELTA-isomerase, SDS-PAGE
26000 - 68000
-
protein concentration 0.043 mg/ml-15.6 mg/ml. At concentrations below 1 mg/ml the enzyme exists as a dimer of MW 26800, determined by gel filtration and ultracentrifugation studies. Concentration-dependent association occurs at higher enzyme concentrations. The latter value represents the weight avarage molecular weight of two or more polymerization species in rapid equilibrium, rather than a discrete polymeric form of the enzyme
300000
-
DELTA5-3-beta-hydroxysteroid dehydrogenase-isomerase system, gel filtration
40000
-
x * 40000, DELTA5-3-beta-hydroxysteroid dehydrogenase-isomerase system, SDS-PAGE
45000
-
2 * 45000, 3beta-hydroxy-5-ene steroid dehydrogenase/isomerase enzyme system, SDS-PAGE
46800
-
x * 46800, SDS-PAGE, enzyme with steroid DELTA-isomerase activity and 3beta-hydroxy-5-ene-steroid dehydrogenase activity
76000
-
enzyme complex of 3beta-hydroxy-5-ene-steroid dehydrogenase and steroid DELTA-isomerase, gel filtration
77000
-
enzyme complex of 3beta-hydroxy-5-ene-steroid dehydrogenase and steroid DELTA-isomerase gel filtration
81000
-
3beta-hydroxy-5-ene steroid dehydrogenase/steroid DELTA-isomerase system, gel filtration
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
homodimer
tetramer
-
4 * 19000, enzyme complex of 3beta-hydroxy-5-ene-steroid dehydrogenase and steroid DELTA-isomerase, SDS-PAGE
additional information
-
detection of an intermediate during the unfolding process of the dimeric ketosteroid isomerase, NMR spectroscopy, circular dichroism and fluorescence spectroscopies, and small angle X-ray scattering and analytical ultracentrifugation of native and fully unfolded enzymes, overview, the formation of a compact intermediate should precede the association of monomers prior to the dimerization process during the folding of KSI
?
-
x * 40000, DELTA5-3-beta-hydroxysteroid dehydrogenase-isomerase system, SDS-PAGE
?
-
x * 46800, SDS-PAGE, enzyme with steroid DELTA-isomerase activity and 3beta-hydroxy-5-ene-steroid dehydrogenase activity
dimer
-
2 * 45000, 3beta-hydroxy-5-ene steroid dehydrogenase/isomerase enzyme system, SDS-PAGE
dimer
-
2 * 11000, SDS-PAGE. At concentrations below 1 mg/ml the enzyme exists as a dimer of MW 26800, determined by gel filtration and ultracentrifugation studies. Concentration-dependent association occurs at higher enzyme concentrations
dimer
active enzyme, interface is composed of two pairs of helices related by a 2fold symmetry axis, involved residues, overview
dimer
-
2 * 14000, mutant enzymnes R72A, E118A, N120A and E18A/N120A, SDS-PAGE
homodimer
-
131 amino acid residues per monomer, xenon is employed as an indirect probe for multidimensional NMR to investigate the structural changes in the urea-induced equilibrium unfolding
homodimer
-
determined by 3D NMR, the secondary and tertiary structures of KSI are not affected significantly by urea up to 3.5 M. The chemical shift analysis of 1H-15N HSQC spectra at various urea concentrations reveales that the residues in the dimeric interface region, particularly around the beta5-strand, are significantly perturbed by urea at low concentrations, while the line-width analysis indicates the possibility of conformational exchange at the interface region around the beta6-strand. The interface region primarily around the beta5- and beta6-strands could play an important role as the starting positions in the unfolding process of KSI.
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
1.2-1.5 A resolution X-ray crystallography, 1H and 19F NMR spectroscopy, quantum mechanical calculations, and transition-state analogue binding measurements of the active site. Packing and binding interactions within the KSI active site can constrain local side-chain reorientation and prevent hydrogen bond shortening by 0.1 A or less. This constraint has substantial energetic effects on ligand binding and stabilization of negative charge within the oxyanion hole. Structural features of the oxyanion hole suggest that hydrogen bond formation to the reacting substrate is geometrically optimal in the transition state but not in the ground state. During steroid isomerization, the hybridization of the substrate oxygen changes from a planar sp2 carbonyl to a tetrahedral sp3 dienolate, altering the spatial distribution of its lone pair electrons. This reorientation of atomic orbitals about the substrate oxygen alters its geometric preference for accepting hydrogen bonds
-
crystals are grown at 12°C from1.3 M ammonium sulfate, 3-5% poly(ethylene glycol)400 and 0.1 M HEPES, pH 7.5 in hanging drops. The crystal structure at 2.3 resolution reveals that the active site environment of the Comamonas testosteroni enzyme is nearly identical to that of Pseudomonas putida enzyme
crystals of mutant enzyme F116W are grown by hanging-drop method
study on backbone dynamics in free enzyme and its complex with a steroid analogue, 19-nortestosterone hemisuccinate. Mutation Y14F induces a substantial decrease in the order parameters in free enzyme, indicating that the backbone structures become significantly mobile by mutation, while the chemical shift analysis indicates that the structural perturbations are more profound than those of wild-type upon 19-nortestosterone hemisuccinate binding. In the 19-nortestosterone hemisuccinate complexed mutant, the key active site residues including Tyr14, Asp38 and Asp99 or the regions around them remain flexible with significantly reduced S2 values, whereas the S2 values for many of the residues in the mutant enzyme become even greater than those of wild-type
-
The 2.26 A crystal structure of the enzyme in complex with a reaction intermediate analogue equilenin reveals that both the Tyr14 OH and the Asp99 COOH provide direct hydrogen bonds to the oxyanion of equilenin
GST A3-3 in complex with DELTA5-androstene-3,17-dione, using 100 mM Tris-HCl pH 7.8, 18% (v/v) PEG 4000, and 2 mM dithiothreitol
hanging-drop vapor diffusion method. The inability to crystallize the detergent-solubilized, wild-type microsomal enzyme is overcome by engineering a cytosolic form of this protein. Modified enzyme in which the membrane-spanning domain, residues 283-310 of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells
-
identification of potentially critical residues M187 and S124 by docking of trilostane or 4alpha,5alpha-epoxy-testosterone into the active site
-
1.2-1.5 A resolution X-ray crystallography, 1H and 19F NMR spectroscopy, quantum mechanical calculations, and transition-state analogue binding measurements of the active site. Packing and binding interactions within the KSI active site can constrain local side-chain reorientation and prevent hydrogen bond shortening by 0.1 A or less. This constraint has substantial energetic effects on ligand binding and stabilization of negative charge within the oxyanion hole. Structural features of the oxyanion hole suggest that hydrogen bond formation to the reacting substrate is geometrically optimal in the transition state but not in the ground state. During steroid isomerization, the hybridization of the substrate oxygen changes from a planar sp2 carbonyl to a tetrahedral sp3 dienolate, altering the spatial distribution of its lone pair electrons. This reorientation of atomic orbitals about the substrate oxygen alters its geometric preference for accepting hydrogen bonds
crystal structure of mutant enzyme F82A is determined to 2.1 A resolution. Crystals are grown in a solution containing 1.0 M sodium acetate and 0.1 M ammonioum acetate, pH 4.6, by the hanging drop method of vapor diffusion at 22 C. The crystals belong to the space group c2221 with unit cell dimensions of a = 36.24 A, b = 96.13 A and c = 74.30 A
-
crystal structure of the enzyme in complex with equilenin, an analogue of the reaction intermediate at 1.9 and 2.5 A resolution
crystal structure of the R72A mutant enzyme determined at 2.5 A resolution belongs to the space group C2221 with cell dimensions of a = 36.37 A, b = 74.44 A and c = 96.06 A. Crystals are grown from a solution containing 2.0 M ammonioum acetate and 0.1 M sodium acetate at pH 4.6 by hanging drop vapor-diffusion method at 22°C
-
crystals of Y30F, Y55F, and Y30F/Y55F are grown in the solution containing 1.0 M sodium acetate and 0.1 M ammonium acetate, pH 4.6 by hanging drop method of vapor diffusion at 22°C. The crystal structure of Y55F as determined at 1.9 A resolution shows that Tyr14 OH undergoes an alteration in orientation to form a new hydrogen bond with Tyr30
-
enzyme mutant D40N bound to phenolate, X-ray diffraction structure determination and analysis at 1.25 A resolution
-
hanging drop vapor diffusion method. Crystal structures of Y14F and Y14F/Y30F/Y55F are determined at 1.8 and 2.0 A resolution, respectively
-
mutant D103N/D40N bound to inhibitor equilenin, hanging drop vapor diffusion method, 0.002 ml of 25 mg/ml protein with equilenin in a molar ratio of 1:1.2 in 40 mM potassium phosphate, pH 7.2, are mixed with 0.002 ml of reservoir solution conraining 1.4 M ammonium sulfate, and 6.5% v/v 2-propanol, pH 7.0, room temperature, 1 week, X-ray diffraction structure determination and analysis at 1.1 A resolution
mutant enzyme D99E/D38N complexed with equilenin, an intermediate analogue, crystals of the complexes are grown from 1.1 M ammonium acetate and 0.1 M sodium acetate, pH 4.6, by the hanging drop vapor diffusion method at 22°C. The resulting crystals have C2 space group symmetry with unit cell dimensions of a = 89.04 A, b = 72.42 A, c = 51.24 A and beta = 90.9°
-
mutant enzyme Y30F/Y55F/Y115F/D38N KSI complexed with equilenin, hanging drop vapor diffusion method, using 0.1 M sodium acetate, pH 4.5, 0.6 M ammonium acetate, and 30% PEG 4000
mutant W92A, in complex with d-equilenin, decrease in conformational stability results from destabilization of surface hydrophobic layer
-
mutant Y14F/D99L, increase in hydrophobic interaction while disrupting the hydrogen bond network, mutants Y30F/D99L and Y55F/D99L, disruption of hydrogen bond network
-
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D103A
-
site-directed mutagenesis of the catalytic residue, the mutant shows 5000fold reduced activity compared tot he wild-type enzyme
D38A
-
the catalytic turnover number is 140fold less than that for the wild-type
D38E
D40N
-
site-directed mutagenesis, the mutation mimics the protonated aspartate found in the intermediate and equilenin complexes and leads to tighter binding of phenolate and other intermediate analogs
D99A
-
secondary kinetic isotope effects similar to wild-type. Mutation does not significantly decrease the contribution of coupled motion/hydrogen tunneling to the enzymatic reaction
D99L
the mutant shows about 10000fold decreased kcat compared to the wild type enzyme
F116W
the turnover-number for 5-androstene-3,17-dione is lowered 4.42fold, the KM-value is 3.1fold lower than the Km-value of the wild-type enzyme
Y14F
Y14F/D99L
the mutant shows 1000000fold decreased kcat compared to the wild type enzyme
Y16F
-
site-directed mutagenesis of the catalytic residue, the mutant shows 50000fold reduced activity compared tot he wild-type enzyme
Y55F/Y88F
-
replacement of Tyr14 by 3-fluorotyrosine in the Y55,88F modified form of the isomerase results in a 4-fold decrease in turnover number
Y57F/D40N
-
site-directed mutagenesis, 3,4,5-trifluorophenol bind as ionized phenolate to KSI containing the Y57F mutation
D241N
-
mutant enzyme nearly has full isomerase activity, substantial decrease in 3beta-hydroxysteroid dehydrogenase activity. Mutant enzyme has a basal isomerase activity in the absence of coenzyme that is 10% of the NADH-stimulated turnover number
D257L
-
complete lack of isomerase activity in absence of coenzyme, substantial decrease in 3beta-hydroxysteroid dehydrogenase activity
D258L
-
complete lack of isomerase activity in absence of coenzyme, substantial decrease in 3beta-hydroxysteroid dehydrogenase activity
D265N
-
complete lack of isomerase activity in absence of coenzyme, substantial decrease in 3beta-hydroxysteroid dehydrogenase activity
D36A/K37R
-
mutation shifts the cofactor preference of both 3-beta-hydroxysteroid dehydrogenase and isomerase from NAD(H) to NADP(H)
DELTA283-310
-
the molecular weight of the subunit is 38800 Da compared to 42000 Da for the wild-type enzyme
E126L
site-directed mutagenesis, altered kinetics compared to the wild-type enzyme
H156Y
site-directed mutagenesis, leads to destabilization of interactions at the dimer interface and a dramatic increase in the substrate Km and inhibitor Ki values of 3beta-HSD isozyme 1 to equal those measured for 3beta-HSD isozyme 2
H232A
site-directed mutagenesis, altered kinetics compared to the wild-type enzyme
K158Q
site-directed mutagenesis, results in the complete loss of dehydrogenase activity and a reduction in isomerase activity, probably due to partial destabilization of cofactor binding, in which the catalytic triad is actively involved, and associated substrate binding
N100A
site-directed mutagenesis, altered kinetics compared to the wild-type enzyme
N100S
site-directed mutagenesis, altered kinetics compared to the wild-type enzyme
N323L
site-directed mutagenesis, altered kinetics compared to the wild-type enzyme
Q105M
site-directed mutagenesis, leads to destabilization of interactions at the dimer interface and a dramatic increase in the substrate Km and inhibitor Ki values of 3beta-HSD isozyme 1 to equal those measured for 3beta-HSD isozyme 2
S124A
site-directed mutagenesis, results in the complete loss of dehydrogenase activity and a reduction in isomerase activity, probably due to partial destabilization of cofactor binding, in which the catalytic triad is actively involved, and associated substrate binding
S322A
site-directed mutagenesis, altered kinetics compared to the wild-type enzyme
Y154F
site-directed mutagenesis, results in the complete loss of dehydrogenase activity and a reduction in isomerase activity, probably due to partial destabilization of cofactor binding, in which the catalytic triad is actively involved, and associated substrate binding
D103N/D40N
site-directed mutagenesis, 3,4,5-trifluorophenol bind as ionized phenolate to KSI containing the D103N mutation
D40N
D99E
-
turnover-number for 5-androstene-3,17-dione is 68.1% of the turnover number of the wild-type enzyme, the Km-value is 1.22fold higher than the Km-value of the wild-type enzyme
D99L
E118A
-
the free-energy change for unfolding in the absence of urea at 25°C is decreased by about 3.9 kcal/mol compared to wild-type value. Mutation increasex the dissociation constant for (+)-equilenin, a reaction intermediate analogue. 50% of the protein is unfolded at 4.46 M urea compared to 5.22 M for the wild-type enzyme. The turnover-number for 5-androstene-3,17-dione is 33% of that of the wild-type enzyme, the Km-value is 348% of that of the wild-type enzyme
E118A/N120A
-
the free-energy change for unfolding in the absence of urea at 25°C is decreased by about 9.5 kcal/mol compared to wild-type value. Mutation increases the dissociation constant for (+)-equilenin, a reaction intermediate analogue. 50% of the protein is unfolded at 3.89 M urea compared to 5.22 M for the wild-type enzyme. The turnover-number for 5-androstene-3,17-dione is 6% of that of the wild-type enzyme, the Km-value is 523% of that of the wild-type enzyme
F54A
-
turnover number for 5-androstene-3,17-dione is 17.1% of the turnover number for the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 15.3% of the turnover number of the wild-type enzyme,the KM-value for 5-androstene-3,17-dione is 116% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 233% of the KM-value of the wild-type enzyme
F54L
-
turnover number for 5-androstene-3,17-dione is 41.9% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 38.7% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 154% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 89.3% of the KM-value of the wild-type enzyme
F82A
-
turnover number for 5-androstene-3,17-dione is 2.5% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 2.3% of the turnover number of the wild-type enzyme,the KM-value for 5-androstene-3,17-dione is 29.9% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 29.8% of the KM-value of the wild-type enzyme
F82L
-
turnover number for 5-androstene-3,17-dione is 12% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 67.6% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 108% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 130% of the KM-value of the wild-type enzyme
L125A/V127A
-
mutation in small exterior hydrophobic cluster, decrease in conformational stability
L125F/V127F
-
mutation in small exterior hydrophobic cluster, slight increase in stability
N120A
-
the free-energy change for unfolding in the absence of urea at 25°C is decreased by about 7.8 kcal/mol compared to wild-type value. Mutation increasex the dissociation constant for (+)-equilenin, a reaction intermediate analogue. 50% of the protein is unfolded at 3.95 M urea compared to 5.22 M for the wild-type enzyme. The turnover-number for 5-androstene-3,17-dione is 17% of that of the wild-type enzyme, the Km-value is 516% of that of the wild-type enzyme
R72A
-
the free-energy change for unfolding in the absence of urea at 25°C is decreased by about 3.8 kcal/mol compared to wild-type value. Mutation increasex the dissociation constant for (+)-equilenin, a reaction intermediate analogue. 50% of the protein is unfolded at 4.74 M urea compared to 5.22 M for the wild-type enzyme. The turnover-number for 5-androstene-3,17-dione is 23% of that of the wild-type enzyme
W116A
-
turnover number for 5-androstene-3,17-dione is 1.5% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is below 12%% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 348% of the KM-value of the wild-type enzyme
W116F
-
turnover number for 5-androstene-3,17-dione is 22.4% of the turnover number of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 299% of the KM-value of the wild-type enzyme
W116Y
-
turnover number for 5-androstene-3,17-dione is 21% of the turnover number of the wild-type enzyme
W92A
-
mutation in small exterior hydrophobic cluster, decrease in conformational stability. Crystallization data
W92A/L125A/V127A
-
mutation in small exterior hydrophobic cluster, decrease in conformational stability
Y115F
the mutant shows about 68% activity compared to the wild type enzyme
Y14F
Y14F/D99E
-
turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.05fold lower than the Km-value of the wild-type enzyme
Y14F/D99L
Y14F/D99N
-
turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.06fold lower than the Km-value of the wild-type enzyme
Y14F/Y30F
-
the turnover-number for 5-androstene-3,17-dione is about 1% of that of the wild-type enzyme, the Km-value is 1.57fold higher than that of the wild-type enzyme
Y14F/Y30F/Y55F
-
the turnover-number for 5-androstene-3,17-dione is about 2.1% of that of the wild-type enzyme, the Km-value is comparable to the Km-value of the wild-type enzyme. The hydrogen bond between Asp99 Odelta2 and C3-O of the steroid, which is perturbed by the Y14F mutation, can be partially restored to that in the wild-type enzyme by additional Y30F/Y55F mutations. The improvement in the catalytic activity of Y14F by the additional Y30f/Y55F mutation is due to the changes in the structural integrity at the catalytic site and the resulting restoration of the proton-transfer mechanism in Y14F/Y30F/Y55F
Y14F/Y30F/Y55F/D99L
-
the turnover-number for 5-androstene-3,17-dione is less than 1.2% of that of the wild-type enzyme, the Km-value is 1.8fold higher than that of the wild-type enzyme
Y14F/Y30F/Y55F/D99N
-
the turnover-number for 5-androstene-3,17-dione is less than 1% of that of the wild-type enzyme, the Km-value is 1.14fold higher than that of the wild-type enzyme
Y14F/Y55F
-
the turnover-number for 5-androstene-3,17-dione is about 1.2% of that of the wild-type enzyme, the Km-value is 1.75fold higher than that of the wild-type enzyme
Y16F
Y16F/Y32F/Y57F
the number of water molecules directly hydrogen bonded to the ligand oxygen is one in the Y16F/Y32F/Y57F mutant
Y16S
the number of water molecules directly hydrogen bonded to the ligand oxygen is approximately two in the Y16S mutant
Y30F
Y30F/D99L
Y30F/Y55F
Y30F/Y55F/Y115F
the mutant shows about 57% activity compared to the wild type enzyme
Y55F
Y55F/D99L
Y57S
-
the mutation causes a large decrease in the catalytic performance of the enzyme
D38E
compared to wild-type, 200fold reduction in kcat-value. Analysis of thermodynamic parameters
D38E
-
the mutant gives similar free energies to the native enzyme, with catalytic constants approximately 200-300times less than in wild type enzyme
Y14F
-
secondary kinetic isotope effects similar to wild-type. Mutation does not significantly decrease the contribution of coupled motion/hydrogen tunneling to the enzymatic reaction
Y14F
-
study on backbone dynamics in free enzyme and its complex with a steroid analogue, 19-nortestosterone hemisuccinate. Mutation induces a substantial decrease in the order parameters in free enzyme, indicating that the backbone structures become significantly mobile by mutation, while the chemical shift analysis indicates that the structural perturbations are more profound than those of wild-type upon 19-nortestosterone hemisuccinate binding. In the 19-nortestosterone hemisuccinate complexed mutant, the key active site residues including Tyr14, Asp38 and Asp99 or the regions around them remain flexible with significantly reduced S2 values, whereas the S2 values for many of the residues in the mutant enzyme become even greater than those of wild-type
Y14F
the mutant shows 100000fold decreased kcat compared to the wild type enzyme
D40N
-
1484555fold decrease in turnover number, 4.46fold decrease in KM-value as compared to wild-type enzyme
D40N
-
site-directed mutagenesis, the mutation mimics the protonated aspartate found in the intermediate and equilenin complexes and leads to tighter binding of phenolate and other intermediate analogs
D40N
-
the mutation does not perturb the gross tertiary and secondary structure of the enzyme The D40N mutant mimics the charge distribution of the enzyme active site in the intermediate state (protonated base), and is not the active form of the enzyme
D99L
-
the turnover number for 5-androstene-3,17-dione is 96.5fold lower than that of the wild-type enzyme, the KM-value is 1.9fold lower than that of the wild-type enzyme. Mutation results in a loss of conformational stability of 3.8 kcal/mol, at 25°C, pH 7.0
D99L
-
turnover-number for 5-androstene-3,17-dione is 125fold lower than the turnover number of the wild-type enzyme, the Km-value is 1.95fold lower than the Km-value of the wild-type enzyme
Y14F
-
mutation results in a loss of conformational stability of 4.4 kcal/mol, at 25°C, pH 7.0
Y14F
-
the hydrogen bond between Asp99 Odelta2 and C3-O of the steroid, which is perturbed by the Y14F mutation, can be partially restored to that in the wild-type enzyme by additional Y30F/Y55F mutations
Y14F
-
turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.87fold lower than the Km-value of the wild-type enzyme
Y14F
the mutant shows almost no activity compared to the wild type enzyme
Y14F/D99L
-
turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.95fold higher than the Km-value of the wild-type enzyme
Y14F/D99L
-
partly additive effect of mutations for both, catalysis and stability, increase in hydrophobic interaction while disrupting the hydrogen bond network, crystallization data
Y16F
-
2009fold decrease in turnover number, 3.5fold decrease in KM-value as compared to wild-type enzyme
Y16F
the number of water molecules directly hydrogen bonded to the ligand oxygen is one in the Y16F mutant
Y30F
-
the turnover number for 5-androstene-3,17-dione is 1.2fold lower than that of the wild-type enzyme, the KM-value is 1.1fold lower than that of the wild-type enzyme
Y30F
the mutant shows about 84% activity compared to the wild type enzyme
Y30F/D99L
-
the turnover number for 5-androstene-3,17-dione is 521.6fold lower than that of the wild-type enzyme, the KM-value is 1.47fold lower than that of the wild-type enzyme. Mutation results in a loss of conformational stability of 6.5 kcal/mol, at 25°C, pH 7.0
Y30F/Y55F
-
the turnover number for 5-androstene-3,17-dione is 1.99fold lower than that of the wild-type enzyme, the KM-value is comparable to that of the wild-type enzyme
Y30F/Y55F
the mutant shows about 50% activity compared to the wild type enzyme
Y55F
-
the turnover number for 5-androstene-3,17-dione is 6fold lower than that of the wild-type enzyme, the KM-value is 2.17fold lower than that of the wild-type enzyme. mutation results in a loss of conformational stability of 3.5 kcal/mol, at 25°C, pH 7.0. The crystal structure of Y55F as determined at 1.9 A resolution shows that Tyr14 OH undergoes an alteration in orientation to form a new hydrogen bond with Tyr30
Y55F
the mutant shows about 17% activity compared to the wild type enzyme
Y55F/D99L
-
the turnover number for 5-androstene-3,17-dione is 17692fold lower than that of the wild-type enzyme, the KM-value is 1.24fold lower than that of the wild-type enzyme. Mutation results in a loss of conformational stability of 7.9 kcal/mol, at 25°C, pH 7.0
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
-
the free-energy change for unfolding in the absence of urea at 25°C is 24.4 kcal/mol for the wild-type enzyme
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
active-site-directed photoinactivation is sensitized by unsaturated steroid ketone photoaffinity reagents
-
in the equilibrium unfolding process the enzyme stability increases by 2.5 kcal/mol in presence of 5% 2,2,2-trifluoroethanol, the increase in entropy by 2,2,2-trifluoroethanol is partially responsible for the increase in stability
-
photoinactivation by ultraviolet irradiation in the presence of the solid-phase photoaffinity reagent DELTA6-testosterone agarose
-
ultraviolet light-dependent photoinactivation is stimulated by DELTA4-3-ketosteroids. The loss of enzymic activity can be correlated with destruction of a single Asp or Asn
-
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
urea
-
the recovery of the activity by refolding is not diminished even after a prolonged time, 24 h of exposure to 7 M urea. Reversibility of the folding is assessed by a 100fold dilution of the denatured protein and subsequent determination of enzyme activity. The refolding kinetics as monitored by fluorescence intensity can be described as a fast first-order process followed by a second-order and a subsequent slow first-order processes, there may be a monomeric folding intermediate
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
15N labelled KSI is purified from Escherichia coli BL21 DE3 grown in minimal medium (M9) containing 15N ammonium chloride as the nitrogen source.
-
3beta-hydroxy-5-ene-steroid dehydrogenase/steroid DELTA5-DELTA4-ene isomerase enzyme complex
-
ammonium sulfate precipitation and phenyl Sepharose 6 column chromatography
-
enzyme complex of 3beta-hydroxy-5-ene-steroid dehydrogenase and steroid 5 to 4 isomerase
-
modified enzyme in which the membrane-spanning domain, residues 283-310, of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells
-
mutant enzymes D36A/K37R, D241N, D257L, D258L and D265N are expressed in a baculovirus/Sf9 cell system
-
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BLR(DE3) to homogeneity by expanded bed adsorption column utilizing immobilized metal affinity chromatography with denaturant concentration in the feed stream from 8 to 0 M urea, chemical extraction from cells, method optimization and validation, overview
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BLR(DE3) to homogeneity by expanded bed adsorption column utilizing immobilized metal affinity chromatography, this integrated process greatly simplifies the recovery and purification of inclusion body proteins by removing the need for mechanical cell disruption, repeated inclusion body centrifugation, and difficult clarification operations, chemical extraction is as effective as homogenization at releasing the inclusion body proteins from the bacterial cells, over 99% purity of the enzyme, method optimization and validation, overview
recombinant enzyme from Escherichia coli strain BLR(DE3) to homogeneity by nickel affinity chromatography after dilution refolding, method optimization, overview
recombinant type 1 isozyme, wild-type and mutant enzymes, from Spodoptera frugiperda Sf9 cells
recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha to over 99% purity
the 3beta-hydroxysteroid dehydrogenase cannot be separated from the isomerase
-
recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha to over 99% purity
-
recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha to over 99% purity
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
D38A, D38A/Y14F, D38A/Y55F and D38A/D99N proteins are expressed in Epicurian Coli XL1-Blue cells
-
expression of C-terminally His6-tagged enzyme as insoluble protein in inclusion bodies in Escherichia coli strain BLR(DE3)
expression of type 1 isozyme, wild-type and mutant enzymes, in Spodoptera frugiperda Sf9 cells via baculovirus infection
expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha
mutant enzymes D36A/K37R, D241N, D257L, D258L and D265N are expressed in a baculovirus/Sf9 cell system
-
expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha
-
expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
during the estrous cycle, the endometrial enzyme activity is higher on days 10 to 11 and decreases on days 12 to 13 and days 15 to 16
-
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RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
50% of the wild-type enzyme is unfolded at 5.22 M urea. Mutant enzymes R72A, E118A, N120A and E118A/N120A are unfolded at lower concentrations
-
70-87% refolding of recombinant C-terminally His6-tagged enzyme after expression in Escherichia coli as insoluble protein and solubilization by denaturation, detergent treatment, and 40fold dilution by a single-step column-based refolding step with an elution gradient, dilution with buffer 1% bovine serum albumin, 20 mM phosphate, 150 mM NaCl, optimal at pH 6.8, method optimization, overview
detection of an intermediate during the unfolding process of the dimeric ketosteroid isomerase, NMR spectroscopy, circular dichroism and fluorescence spectroscopies, and small angle X-ray scattering and analytical ultracentrifugation of native and fully unfolded enzymes, overview
-
dilution refolding of recombinant enzyme from inclusion bodies, denatured recombinant C-terminally His6-tagged enzyme 10fold with refolding buffer containing 50 mM potassium phosphate, pH 7.0, 4°C, incubation at 4°C for 12 h, urea 0.8 M
repeated adsorptive refolding in an expanded bed with gradient change in the feed-stream composition from denaturing buffer containing 8 M urea and 50 mM potassium phosphate, pH 6.8 to the refolding buffer containing 50 mM potassium phosphate, pH 6.8, constant liquid feed rate, method evaluation, overview
secondary structure is not significantly affected by urea up to 3.5 M, but the residues in the dimeric interface region are significantly perturbed by urea at low concentrations. The interface region primarily around the beta5- and beta6-strands may play an important role as the starting positions in the unfolding process
-
the recovery of the activity by refolding is not diminished even after a prolonged time, 24 h of exposure to 7 M urea
-
urea-induced equilibrium unfolding, two-state mechanism involving only the native dimer and the unfolded monomer. The enzymatic activity is affected significantly by urea and decreases exponentially as urea concentration increases, suggesting that the active site is less stable than the tertiary structure as a whole. Dilution of the unfolded protein in 8 M urea to lower urea concentrations in a reducing condition, the activity of the refolded protein is recovered up to over 95% of that of the native protein. The protein folds into a monomer containing most of the alpha-helical structures before dimerization
-
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kawahara, F.S.; Wang, S.F.; Talalay, P.
The preparation and properties of crystalline DELTA5-3-ketosteroid isomerase
J. Biol. Chem.
237
1500-1506
1962
Comamonas testosteroni
brenda
Jones, J.B.; Ship, S.
Inhibition of the 3-ketosteroid DELTA5->DELTA4 isomerase of Pseudomonas testosteroni by some bromo-3-ketosteroid derivatives
Biochim. Biophys. Acta
258
800-809
1972
Comamonas testosteroni
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Comamonas testosteroni
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The identification, properties, and immobilization of the 3beta-hydroxysteroid oxidase and 3-oxosteroid DELTA4-DELTA5-isomerase components of cholesterol oxidase from Nocardia rhodochrous
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Comamonas testosteroni
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Characterization of a hexagonal crystal form of an enzyme of steroid metabolism, DELTA5-3-ketosteroid isomerase: a new method of crystal density measurement
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Comamonas testosteroni
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Interaction of steroids with Pseudomonas testosteroni 3-oxosteroid DELTA4-DELTA5 isomerase
Biochemistry
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1977
Comamonas testosteroni
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The substrate specificity and stereochemistry, reversibility and inhibition of the 3-oxo steroid DELTA 4-DELTA 5-isomerase component of cholesterol oxidase
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1981
Comamonas testosteroni
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Irreversible inactivation of DELTA5-3-ketosteroid isomerase of Pseudomonas testosteroni by acetylenic suicide substrates. Mechanism of formation and properties of the steroid-enzyme adduct
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1981
Comamonas testosteroni
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Use of a solid-phase photoaffinity reagent to label a steroid binding site: application to the DELTA5-3-ketosteroid isomerase of Pseudomonas testosteroni
Biochemistry
24
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1985
Comamonas testosteroni
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Enzymic isomerization of DELTA5-3-ketosteroids
Biochim. Biophys. Acta
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1955
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1976
Comamonas testosteroni
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Simard, J.; Melner, M.H.; Breton, N.; Low, K.G.; Zhao, H.F.; Periman, L.M.; Labrie, F.
Characterization of macaque 3beta-hydroxy-5-ene steroid dehydrogenase/DELTA5-DELTA4-isomerase: structure and expression in steroidogenic and peripheral tissues in primate
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1991
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Mechanism of the reaction catalyzed by DELTA5-3-ketosteroid isomerase of Comamonas (Pseudomonas) testosteroni: kinetic properties of a modified enzyme in which tyrosine 14 is replaced by 3-fluorotyrosine
Biochemistry
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Comamonas testosteroni
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pH-Dependence of the kinetic parameters for 3-oxo-DELTA5-steroid isomerase substrate catalysis and inhibition by (3S)-spiro[5alpha-androstane-3,2'-oxiran]-17-one
Biochemistry
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1986
Comamonas testosteroni
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Bevins, C.L.; Pollack, R.M.; Kayser, R.H.; Bounds, P.L.
Detection of transient enzyme-steroid complex during active-site-directed irreversible inhibition of 3-oxo-DELTA5-steroid isomerase
Biochemistry
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1986
Comamonas testosteroni
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Purification and characterization of 3beta-hydroxysteroid-dehydrogenase/isomerase from bovine adrenal cortex
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Bos taurus
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Characterization, expression, and immunohistochemical localization of 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase in human skin
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Homo sapiens
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1996
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1997
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Crystal structure and enzyme mechanism of Delta 5-3-ketosteroid isomerase from Pseudomonas testosteroni
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Comamonas testosteroni
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3beta-Hydroxy-DELTA5-steroid dehydrogenase/3-keto-DELTA5-steroid isomerase from bovine adrenals: mechanism of inhibition by 3-oxo-4-aza steroids and kinetic mechanism of the dehydrogenase
Biochemistry
28
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1989
Bos taurus
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Affinity alkylation of bacterial DELTA5-3-ketosteroid isomerase. Identification of the amino acid modified by steroidal 17beta-oxiranes
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1983
Comamonas testosteroni
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Covey, D.F.; Robinson, C.H.
Conjugated allenic 3-oxo-5,10-secosteroids. Irreversible inhibitors of DELTA5-3-ketosteroid isomerase
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1976
Comamonas testosteroni
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1975
Comamonas testosteroni
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Hearne, M.; Benisek, W.F.
Modification of DELTA5-3-ketosteroid isomerase induced by ultraviolet irradiation in the presence of the solid-phase photoaffinity reagent DELTA6-testosterone agarose
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1984
Comamonas testosteroni
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Ishii-Ohba, H.; Inano, H.; Tamaoki, B.
Purification and properties of testicular 3beta-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase
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1986
Rattus norvegicus
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Roles of active site aromatic residues in catalysis by ketosteroid isomerase from Pseudomonas putida biotype B
Biochemistry
38
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1999
Pseudomonas putida
brenda
Kim, D.H.; Jang, D.S.; Nam, G.H.; Yun, S.; Cho, J.H.; Choi, G.; Lee, H.C.; Choi, K.Y.
Equilibrium and kinetic analysis of folding of ketosteroid isomerase from Comamonas testosteroni
Biochemistry
39
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2000
Comamonas testosteroni
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Henot, F.; Pollack, R.M.
Catalytic activity of the D38A mutant of 3-oxo-DELTA 5-steroid isomerase: recruitment of aspartate-99 as the base
Biochemistry
39
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2000
Comamonas testosteroni
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Contribution of the hydrogen-bond network involving a tyrosine triad in the active site to the structure and function of a highly proficient ketosteroid isomerase from Pseudomonas putida biotype B
Biochemistry
39
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2000
Pseudomonas putida
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Choi, G.; Ha, N.C.; Kim, S.W.; Kim, D.H.; Park, S.; Oh, B.H.; Choi, K.Y.
Asp-99 donates a hydrogen bond not to Tyr-14 but to the steroid directly in the catalytic mechanism of DELTA 5-3-ketosteroid isomerase from Pseudomonas putida biotype B
Biochemistry
39
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2000
Pseudomonas putida
brenda
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Pseudoreversion of the catalytic activity of Y14F by the additional substitution(s) of tyrosine with phenylalanine in the hydrogen bond network of DELTA 5-3-ketosteroid isomerase from Pseudomonas putida biotype B
Biochemistry
40
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2001
Pseudomonas putida
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Pseudomonas putida
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Comamonas testosteroni (P00947)
brenda
Yun, S.; Jang do, S.; Choi, G.; Kim, K.S.; Choi, K.Y.; Lee, H.C.
Trifluoroethanol increases the stability of DELTA(5)-3-ketosteroid isomerase. 15N NMR relaxation studies
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Comamonas testosteroni
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Comamonas testosteroni (P00947), Comamonas testosteroni
brenda
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Homo sapiens
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Pseudomonas putida
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Structural double-mutant cycle analysis of a hydrogen bond network in ketosteroid isomerase from Pseudomonas putida biotype B
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Pseudomonas putida
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Comamonas testosteroni
brenda
Soma, K.K.; Alday, N.A.; Hau, M.; Schlinger, B.A.
Dehydroepiandrosterone metabolism by 3beta-hydroxysteroid dehydrogenase/D5-D4 isomerase in adult zebra finch brain: sex difference and rapid effect of stress
Endocrinology
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2004
Taeniopygia guttata
brenda
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Neurosteroids and female reproduction: Estrogen increases 3beta-HSD mRNA and activity in rat hypothalamus
Endocrinology
146
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2005
Rattus norvegicus (P22071)
brenda
Yun, Y.S.; Nam, G.H.; Kim, Y.G.; Oh, B.H.; Choi, K.Y.
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Temperature effects on the catalytic activity of the D38E mutant of 3-oxo-DELTA5-steroid isomerase: favorable enthalpies and entropies of activation relative to the nonenzymatic reaction catalyzed by acetate ion
J. Am. Chem. Soc.
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2004
Comamonas testosteroni (P00947)
brenda
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
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Homo sapiens
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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
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Homo sapiens (P14060), Homo sapiens (P26439), Homo sapiens
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Tam, H.; Schlinger, B.A.
Activities of 3beta-HSD and aromatase in slices of developing and adult zebra finch brain
Gen. Comp. Endocrinol.
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2006
Taeniopygia guttata
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Hutchinson, M.H.; Chase, H.A.
Refolding strategies for ketosteroid isomerase following insoluble expression in Escherichia coli
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Comamonas testosteroni (P00947)
brenda
Hutchinson, M.H.; Morreale, G.; Middelberg, A.P.; Chase, H.A.
Production of enzymatically active ketosteroid isomerase following insoluble expression in Escherichia coli
Biotechnol. Bioeng.
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Comamonas testosteroni (P00947)
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Hutchinson, M.H.; Chase, H.A.
Intensified process for the purification of an enzyme from inclusion bodies using integrated expanded bed adsorption and refolding
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Comamonas testosteroni (P00947)
brenda
Jang, D.S.; Lee, H.J.; Lee, B.; Hong, B.H.; Cha, H.J.; Yoon, J.; Lim, K.; Yoon, Y.J.; Kim, J.; Ree, M.; Lee, H.C.; Choi, K.Y.
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Pseudomonas putida
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Meffre, D.; Delespierre, B.; Gouezou, M.; Schumacher, M.; Stein, D.G.; Guennoun, R.
3beta-Hydroxysteroid dehydrogenase/5-ene-4-ene isomerase mRNA expression in rat brain: effect of pseudopregnancy and traumatic brain injury
J. Steroid Biochem. Mol. Biol.
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Rattus norvegicus (P22071)
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Kraut, D.A.; Sigala, P.A.; Pybus, B.; Liu, C.W.; Ringe, D.; Petsko, G.A.; Herschlag, D.
Testing electrostatic complementarity in enzyme catalysis: hydrogen bonding in the ketosteroid isomerase oxyanion hole
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Comamonas testosteroni, Pseudomonas putida
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Sharma, K.; Vazquez-Ramirez, R.; Kubli-Garfias, C.
A theoretical model of the catalytic mechanism of the DELTA5-3-ketosteroid isomerase reaction
Steroids
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2006
Pseudomonas sp.
brenda
Hunter, A.C.; Coyle, E.; Morse, F.; Dedi, C.; Dodd, H.T.; Koussoroplis, S.J.
Transformation of 5-ene steroids by the fungus Aspergillus tamarii KITA: Mixed molecular fate in lactonization and hydroxylation pathways with identification of a putative 3beta-hydroxy-steroid dehydrogenase/DELTA(5)-DELTA(4) isomerase pathway
Biochim. Biophys. Acta
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2008
Aspergillus tamarii, Aspergillus tamarii KITA
brenda
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Sex- and age-related variation in neurosteroidogenic enzyme mRNA levels during quail embryonic development
Brain Res.
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Coturnix japonica
brenda
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Testing geometrical discrimination within an enzyme active site: constrained hydrogen bonding in the ketosteroid isomerase oxyanion hole
J. Am. Chem. Soc.
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2008
Comamonas testosteroni, Pseudomonas putida (P07445)
brenda
Wilde, T.C.; Blotny, G.; Pollack, R.M.
Experimental evidence for enzyme-enhanced coupled motion/quantum mechanical hydrogen tunneling by ketosteroid isomerase
J. Am. Chem. Soc.
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2008
Comamonas testosteroni
brenda
Lee, H.J.; Yoon, Y.J.; Jang, D.S.; Kim, C.; Cha, H.J.; Hong, B.H.; Choi, K.Y.; Lee, H.C.
15N NMR relaxation studies of Y14F mutant of ketosteroid isomerase: the influence of mutation on backbone mobility
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2008
Comamonas testosteroni
brenda
Lee, H.J.; Jang, do S.; Cha, H.J.; Moon, H.S.; Hong, B.H.; Choi, K.Y.; Lee, H.C.
NMR studies on the equilibrium unfolding of ketosteroid isomerase by urea
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2008
Pseudomonas putida
brenda
Lee, H.J.; Moon, H.S.; Jang, do S.; Cha, H.J.; Hong, B.H.; Choi, K.Y.; Lee, H.C.
Probing the equilibrium unfolding of ketosteroid isomerase through xenon-perturbed (1)H- (15)N multidimensional NMR spectroscopy
J. Biomol. NMR
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65-70
2008
Pseudomonas putida
brenda
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.
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2008
Homo sapiens
brenda
Chakravorty, D.; Soudackov, A.; Hammes-Schiffer, S.
Hybrid quantum/classical molecular dynamics simulations of the proton transfer reactions catalyzed by ketosteroid isomerase: Analysis of hydrogen bonding, conformational motions, and electrostatics
Biochemistry
48
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2009
Comamonas testosteroni (P00947)
brenda
Sigala, P.; Caaveiro, J.; Ringe, D.; Petsko, G.; Herschlag, D.
Hydrogen bond coupling in the ketosteroid isomerase active site
Biochemistry
48
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2009
Comamonas testosteroni, Pseudomonas putida (P07445)
brenda
Childs, W.; Boxer, S.G.
Proton affinity of the oxyanion hole in the active site of ketosteroid isomerase
Biochemistry
49
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2010
Pseudomonas putida
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Childs, W.; Boxer, S.G.
Solvation response along the reaction coordinate in the active site of ketosteroid isomerase
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2010
Pseudomonas putida
brenda
Hanoian, P.; Hammes-Schiffer, S.
Water in the active site of ketosteroid isomerase
Biochemistry
50
6689-6700
2011
Pseudomonas putida (P07445)
brenda
Chakravorty, D.K.; Hammes-Schiffer, S.
Impact of mutation on proton transfer reactions in ketosteroid isomerase: insights from molecular dynamics simulations
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2010
Comamonas testosteroni (P00947)
brenda
Tars, K.; Olin, B.; Mannervik, B.
Structural basis for featuring of steroid isomerase activity in alpha class glutathione transferases
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Homo sapiens (Q16772), Homo sapiens
brenda
Ewald, W.; Werbin, H.; Chaikoff, I.L.
Evidence for the presence of 17-hydroxypregnenedione isomerase in beef adrenal cortex
Biochim. Biophys. Acta
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1965
Bos taurus
brenda
Wojciechowicz, B.; Kotwica, G.; Kolakowska, J.; Franczak, A.
The activity and localization of 3beta-hydroxysteroid dehydrogenase/DELTA(5)-DELTA(4) isomerase and release of androstenedione and progesterone by uterine tissues during early pregnancy and the estrous cycle in pigs
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2013
Sus scrofa
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van der Kamp, M.W.; Chaudret, R.; Mulholland, A.J.
QM/MM modelling of ketosteroid isomerase reactivity indicates that active site closure is integral to catalysis
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Comamonas testosteroni
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Jha, S.K.; Ji, M.; Gaffney, K.J.; Boxer, S.G.
Site-specific measurement of water dynamics in the substrate pocket of ketosteroid isomerase using time-resolved vibrational spectroscopy
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2012
Pseudomonas putida
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Parker, M.; Osuna, S.; Bollot, G.; Vaddypally, S.; Zdilla, M.; Houk, K.; Schafmeister, C.
Acceleration of an aromatic claisen rearrangement via a designed spiroligozyme catalyst that mimics the ketosteroid isomerase catalytic dyad
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2014
Pseudomonas putida (P07445)
brenda
Ito, M.; Brinck, T.
Novel approach for identifying key residues in enzymatic reactions Proton abstraction in ketosteroid isomerase
J. Phys. Chem. B
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2014
Comamonas testosteroni
brenda
Jang, D.S.; Choi, G.; Cha, H.J.; Shin, S.; Hong, B.H.; Lee, H.J.; Lee, H.C.; Choi, K.Y.
Contribution of a low-barrier hydrogen bond to catalysis is not significant in ketosteroid isomerase
Mol. Cells
38
409-415
2015
Pseudomonas putida (P07445)
brenda
Cha, H.; Jang, D.; Jin, K.; Lee, H.; Hong, B.; Kim, E.; Kim, J.; Lee, H.; Choi, K.; Ree, M.
Three-dimensional structures of a wild-type ketosteroid isomerase and its single mutant in solution
Sci. Adv. Mat.
6
2325-2333
2014
Pseudomonas putida
-
brenda
Meitinger, N.; Munkert, J.; Maia de Pdua, R.; de Souza Filho, J.; Maid, H.; Bauer, W.; Braga, F.; Kreis, W.
The catalytic mechanism of the 3-ketosteroid isomerase of Digitalis lanata involves an intramolecular proton transfer and the activity is not associated with the 3beta-hydroxysteroid dehydrogenase activity
Tetrahedron Lett.
57
1567-1571
2016
Digitalis lanata
-
brenda
Currin, A.; Dunstan, M.S.; Johannissen, L.O.; Hollywood, K.A.; Vinaixa, M.; Jervis, A.J.; Swainston, N.; Rattray, N.J.W.; Gardiner, J.M.; Kell, D.B.; Takano, E.; Toogood, H.S.; Scrutton, N.S.
Engineering the missing link in biosynthetic (-)-menthol production bacterial isopulegone isomerase
ACS Catal.
8
2012-2020
2018
Pseudomonas putida (P07445)
brenda
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