Information on EC 1.3.1.3 - DELTA4-3-oxosteroid 5beta-reductase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.3.1.3
-
RECOMMENDED NAME
GeneOntology No.
DELTA4-3-oxosteroid 5beta-reductase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
17,21-dihydroxy-5beta-pregnane-3,11,20-trione + NADP+ = cortisone + NADPH + H+
show the reaction diagram
5beta-cholestan-3-one + NADP+ = cholest-4-en-3-one + NADPH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
reduction
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
bile acid biosynthesis, neutral pathway
Primary bile acid biosynthesis
-
-
Steroid hormone biosynthesis
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
5beta-cholestan-3-one:NADP+ 4,5-oxidoreductase
The enzyme from human efficiently catalyses the reduction of progesterone, androstenedione, 17alpha-hydroxyprogesterone and testosterone to 5beta-reduced metabolites; it can also act on aldosterone, corticosterone and cortisol, but to a lesser extent [8]. The bile acid intermediates 7alpha,12alpha-dihydroxy-4-cholesten-3-one and 7alpha-hydroxy-4-cholesten-3-one can also act as substrates [9].
CAS REGISTRY NUMBER
COMMENTARY hide
37255-35-9
-
9029-08-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
-
deficient 5beta-reductase activity can lead to cholestasis and neo-natal liver failure and is often lethal if it remains untreated
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,4-androstadien-17beta-ol-3-one + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
11-deoxycortisol + NADPH
?
show the reaction diagram
-
-
-
-
?
11beta,17alpha,21-trihydroxy-pregn-4-en-3,20-dione + NADPH
11beta,17alpha,21-trihydroxypregnan-3,20-dione + NADP+
show the reaction diagram
17alpha,21-dihydroxy-pregn-4-en-3,20-dione + NADPH
17alpha,21-dihydroxy-pregnan-3,20-dione + NADP+
show the reaction diagram
17alpha-hydroxyprogesterone + NADPH
?
show the reaction diagram
-
-
-
-
?
17alpha-methylestr-4-en-3-one + NADPH
17alpha-methylestran-3-one + NADP+
show the reaction diagram
17beta-hydroxyandrost-4-en-3-one + NADPH
17beta-hydroxy-5beta-androstan-3-one + NADP+
show the reaction diagram
17beta-hydroxyestr-4-en-3-one + NADPH
17-hydroxyestran-3-one + NADP+
show the reaction diagram
2-cyclohexen-1-one + NADPH + H+
cyclohexanone + NADP+
show the reaction diagram
4-androstene-3,17-dione + NADPH
5beta-androstan-3,15-dione + NADP+
show the reaction diagram
4-androstene-3,17-dione + NADPH
5beta-androstan-3,17-dione + NADP+
show the reaction diagram
4-androstene-3,17-dione + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
4-cholesten-7alpha-ol-3-one + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
4-estren-17beta-ol-3-one + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
7alpha,12alpha-dihydroxy-4-cholesten-3-one + NADPH
7alpha,12alpha-dihydroxy-5-beta-cholestan-3-one + NADP+
show the reaction diagram
-
-
-
-
?
7alpha,12alpha-dihydroxy-4-cholesten-3-one + NADPH + H+
?
show the reaction diagram
-
-
-
-
-
7alpha,12beta-dihydroxy-4-cholesten-3-one + NADPH
(5beta,7alpha,12beta)-7,12-dihydroxycholestan-3-one + NADPH
show the reaction diagram
-
-
-
?
7alpha-hydroxy-4-cholesten + NADPH
7alpha-hydroxy-5beta-cholestan-3-one + NADP+
show the reaction diagram
-
-
-
?
7alpha-hydroxy-4-cholesten-3-one + NADPH
(5beta,7alpha)-7-hydroxycholestan-3-one + NADP+
show the reaction diagram
-
-
-
?
7alpha-hydroxy-4-cholesten-3-one + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
aldosterone + NADPH
?
show the reaction diagram
aldosterone + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
androst-4-en-3,17-dione + NADPH
5beta-androstan-3,17-dione + NADP+
show the reaction diagram
androstenedione + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
cholest-4-en-3-one + NADPH + H+
5beta-cholestan-3-one + NADP+
show the reaction diagram
-
-
-
-
?
cholestenone + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
cortexone + NADPH
21-trihydroxy-5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
corticosterone + NADPH
?
show the reaction diagram
-
low activity
-
-
?
corticosterone + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
cortisol + NADPH
11beta,17,21-trihydroxy-5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
cortisol + NADPH
?
show the reaction diagram
cortisol + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
cortisone + NADPH
11beta,21dihydroxy-pregnan-3,11,20-trione + NADP+
show the reaction diagram
cortisone + NADPH
5beta-17,21dihydroxy-pregnan-3,11,20-trione + NADP+
show the reaction diagram
-
-
-
-
?
cortisone + NADPH
?
show the reaction diagram
cortisone + NADPH + H+
11beta,21dihydroxy-pregnan-3,11,20-trione + NADP+
show the reaction diagram
-
-
-
-
?
cortisone + NADPH + H+
17,21-dihydroxy-5beta-pregnane-3,11,20-trione + NADP+
show the reaction diagram
-
-
-
-
?
cortisone + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
DELTA 4-cholestene-7alpha,12alpha-diol-3-one + NADPH + H+
5beta-cholestane-7alpha,12alpha-diol-3-one + NADP+
show the reaction diagram
-
a bile acid intermediate
-
-
?
DELTA4-cholesten-7alpha-ol-3-one + NADPH + H+
5beta-cholestan-7alpha-ol-3-one + NADP+
show the reaction diagram
-
a bile acid intermediate
-
-
?
deoxycorticosterone + NADPH
pregnan-21-ol-3,20-dione + NADP+
show the reaction diagram
epitestosterone + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
progesterone + NADPH
5beta-pregnan-3,11,20-trione + NADP+
show the reaction diagram
-
-
-
?
progesterone + NADPH
?
show the reaction diagram
progesterone + NADPH
pregnan-3,20-dione + NADP+
show the reaction diagram
progesterone + NADPH + H+
5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
progesterone + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
testosterone + NADPH
?
show the reaction diagram
-
-
-
-
?
testosterone + NADPH + H+
(5beta,17beta)-17-hydroxyandrostan-3-one + NADP+
show the reaction diagram
testosterone + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
17alpha-hydroxyprogesterone + NADPH
?
show the reaction diagram
-
-
-
-
?
2-cyclohexen-1-one + NADPH + H+
cyclohexanone + NADP+
show the reaction diagram
4-androstene-3,17-dione + NADPH
5beta-androstan-3,17-dione + NADP+
show the reaction diagram
aldosterone + NADPH
?
show the reaction diagram
-
low activity
-
-
?
cholest-4-en-3-one + NADPH + H+
5beta-cholestan-3-one + NADP+
show the reaction diagram
-
-
-
-
?
corticosterone + NADPH
?
show the reaction diagram
-
low activity
-
-
?
cortisol + NADPH
?
show the reaction diagram
-
very low activity
-
-
?
cortisone + NADPH + H+
11beta,21dihydroxy-pregnan-3,11,20-trione + NADP+
show the reaction diagram
-
-
-
-
?
cortisone + NADPH + H+
17,21-dihydroxy-5beta-pregnane-3,11,20-trione + NADP+
show the reaction diagram
-
-
-
-
?
DELTA 4-cholestene-7alpha,12alpha-diol-3-one + NADPH + H+
5beta-cholestane-7alpha,12alpha-diol-3-one + NADP+
show the reaction diagram
-
a bile acid intermediate
-
-
?
DELTA4-cholesten-7alpha-ol-3-one + NADPH + H+
5beta-cholestan-7alpha-ol-3-one + NADP+
show the reaction diagram
-
a bile acid intermediate
-
-
?
progesterone + NADPH
?
show the reaction diagram
-
-
-
-
?
progesterone + NADPH + H+
5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
progesterone + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
testosterone + NADPH
?
show the reaction diagram
-
-
-
-
?
testosterone + NADPH + H+
(5beta,17beta)-17-hydroxyandrostan-3-one + NADP+
show the reaction diagram
-
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD(P)H
-
-
additional information
-
NADH: no effect
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
17beta-hydroxy-5alpha-androstan-3-one
-
0.01 mM, 57% inhibition
17beta-hydroxy-5beta-androstan-3-one
-
0.01 mM, 28% inhibition
4-benzoylbenzoic acid
-
-
5beta-dihydrotestosterone
-
product inhibition
androstenedione
chenodeoxycholic acid
Cu2+
-
0.5 mM 66% inhibition
estradiol-17beta
-
0.01 mM, 29% inhibition
estrone
-
0.01 mM, 29% inhibition
Hg2+
-
0.1 mM, 92% inhibition
indomethacin
-
-
iodoacetate
-
-
Mefenamic acid
-
-
N-ethylmaleimide
-
-
NADP+
-
product inhibition
p-chloromercuribenzenesulfonate
-
-
p-chloromercuribenzoate
p-Chloromercuribenzyl sulfonate
-
2 mM, complete inhibition, 50% inhibition in the presence of 10 mM glutathione
progesterone
testosterone
Ursodeoxycholic acid
-
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0032
1,4-androstadien-17beta-ol-3-one
-
pH 6.0, 37C
0.116 - 1.15
2-cyclohexen-1-one
0.0009 - 2.193
4-androstene-3,17-dione
0.0008
4-cholesten-7alpha-ol-3-one
-
pH 6.0, 37C
0.003
4-estren-17beta-ol-3-one
-
pH 6.0, 37C
0.0122
7alpha,12alpha-dihydroxy-4-cholesten-3-one
-
-
0.0091
7alpha-hydroxy-4-cholesten-3-one
-
-
0.0025
aldosterone
-
pH 6.0, 37C
0.022
androst-4-en-3,17-dione
-
-
0.0022
androst-4-ene-3,17-dione
-
-
0.00037 - 0.01629
androstenedione
0.0003
cholestenone
-
pH 6.0, 37C
1.597 - 2.373
cortexone
0.0022
corticosterone
-
pH 6.0, 37C
0.011 - 2.079
cortisol
0.0013 - 0.034
cortisone
0.0029
Epitestosterone
-
pH 6.0, 37C
0.003 - 0.085
NADPH
0.00219 - 0.362
progesterone
0.0016 - 0.0127
testosterone
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.035
1,4-androstadien-17beta-ol-3-one
Homo sapiens
-
pH 6.0, 37C
0.4 - 1.11
2-cyclohexen-1-one
0.0337 - 0.1
4-androstene-3,17-dione
0.033
4-cholesten-7alpha-ol-3-one
Homo sapiens
-
pH 6.0, 37C
0.045
4-estren-17beta-ol-3-one
Homo sapiens
-
pH 6.0, 37C
0.0128
7alpha,12alpha-dihydroxy-4-cholesten-3-one
Rattus norvegicus
-
-
0.01
7alpha-hydroxy-4-cholesten-3-one
Rattus norvegicus
-
-
0.15
aldosterone
Homo sapiens
-
pH 6.0, 37C
0.013 - 0.0425
androstenedione
0.01
cholestenone
Homo sapiens
-
pH 6.0, 37C
0.032
corticosterone
Homo sapiens
-
pH 6.0, 37C
0.00817 - 0.045
cortisol
0.01 - 0.195
cortisone
0.1
Epitestosterone
Homo sapiens
-
pH 6.0, 37C
0.0075 - 1.8
progesterone
0.012 - 1.3
testosterone
additional information
additional information
Sus scrofa
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
10.94
1,4-androstadien-17beta-ol-3-one
Homo sapiens
-
pH 6.0, 37C
42618
0.588 - 9.607
2-cyclohexen-1-one
2446
2.77
4-androstene-3,17-dione
Homo sapiens
-
pH 6.0, 37C
1186
41.25
4-cholesten-7alpha-ol-3-one
Homo sapiens
-
pH 6.0, 37C
42619
15
4-estren-17beta-ol-3-one
Homo sapiens
-
pH 6.0, 37C
42617
60
aldosterone
Homo sapiens
-
pH 6.0, 37C
2825
2.609 - 43.08
androstenedione
1319
33.33
cholestenone
Homo sapiens
-
pH 6.0, 37C
28125
14.54
corticosterone
Homo sapiens
-
pH 6.0, 37C
1103
3.44
cortisol
Homo sapiens
-
pH 6.0, 37C
919
7.69 - 12.91
cortisone
920
34.48
Epitestosterone
Homo sapiens
-
pH 6.0, 37C
2140
0.052 - 0.628
progesterone
286
3.5 - 51.85
testosterone
284
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0092
chenodeoxycholic acid
-
pH not specified in the publication, temperature not specified in the publication
0.00024
progesterone
-
-
0.0145
testosterone
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000036
-
testosterone reduction, enzyme activity in forelimb muscle
0.000167
-
substrate testosterone
0.0153
-
purified enzyme
0.034
-
purified enzyme, substrate cortisone
0.0354
-
purified enzyme, substrate androstenedione
1.89
-
purified wild type enzyme, using progesterone as substrate
13.1
-
purified enzyme, substrate testosterone
additional information
-
105 units/mg, 1 unit is defined as a change in optical density of 0.001 in 20 min
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.4
-
Tris-maleate buffer
6.5
-
androst-4-ene-3,17-dione
7.3
-
assay at
7.8
-
with progesterone as substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2 - 8
-
about 50% of activity maximum at pH 5.2 and 8
5.7 - 8
-
pH 5.7: about 50% of activity maximum, pH 8.0: about 70% of activity maximum
5.8 - 8.2
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
maximally transcribed in mature flowers
Manually annotated by BRENDA team
-
specifically in crustaceans
Manually annotated by BRENDA team
-
at the base of the rosette
Manually annotated by BRENDA team
-
at the basal part of the leaf, lowest amount
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25000 - 30000
-
gel filtration
37000
-
SDS-PAGE, gel filtration
65000
-
recombinant GST-5beta-reductase fusion protein, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
5beta-POR, in the presence and absence of the cofactor NADP+, hanging drop vapour diffusion method, using 15% polyethylene glycol 4000, 0.1 M ammonium acetate, and 0.1 M sodium citrate, pH 5.6
-
by the hanging-drop method, at 2.7 A resolution, crystals belong to space group P43212, contain a single 5beta-POR molecule in the asymmetric unit and tend to diffract anisotropically, identification of six out of eight possible Se-atom positions
-
recombinant wild-type and mutant enzymes from Escherichia coli strain M15 by nickel affinity chromatography
-
the crystal structure of the 5beta-POR-NADP+ binary complex reveals that 5beta-POR exhibits a characteristic short-chain dehydrogenase fold with an N-terminal domain consisting of a double Rossmann fold for cofactor binding and an insertional domain between strands betaF and betaG of about 100 residues for substrate binding, crystal structure of the 5beta-POR-NADP+ binary complex reveals that the side chain of K147 is found in a similar position to the lysine residue of the YXX(S)K motif in standard short-chain dehydrogenases
-
enzyme in binary complex with product NADP+, containing two monomers of AKR1D1, each of which consists of a 325-residue polypeptide chain that adopts an (alpha/beta)8-barrel fold, the AKR1D1-NADP+ complex adopts an extended anti-conformation, X-ray diffraction structure determination and analysis at 1.79 A resolution, comparison with other enzyme binary and tertiary ligand complex structures, overview
-
hanging drop vapour diffusion method in 20% (w/v) PEG-4K, 0.1 M sodium cacodylate (pH 6.5), 0.2 M sodium citrate, 0.5 M (NH4)2SO4
-
hanging drop vapour diffusion method, in 0.1 M Tris/HEPES (pH 7.5), 10-20% (w/v) polyethylene glycol 4000, and 10% isopropyl alcohol
-
purifed recombinant h5beta-red in ternary complex with NADPH and androstenedione, hanging-drop vapor diffusion technique, 2:1 v/v ratio of protein and well solution, about 5 days, X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
-
4C, 30 days, 25% loss of activity
390596
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3
-
complete loss of activity after 24 h
50
-
60 s, 50% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol stabilizes during purification, 4C
-
bovine serum albumin stabilizes
-
highly labile in low buffer concentration
-
markedly stabilized in 20% glycerol
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10C, stable for 2-3 days
-
4C, 72 h, 0.05 M Tris-maleate buffer, pH 7.4-8.4, stable
-
4C, pH 8, 32 days, 25% loss of activity
-
4C, rapid loss of activity, 2-mercaptoethanol stabilizes
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; cortisone 5beta-reduction (EC 1.3.1.3) and androstenedione reduction (EC 1.3.1.23) are catalyzed by the same catalytic site of a single protein
-
by Ni-NTA affinity chromatography
cortisone 5beta-reduction (EC 1.3.1.3) and androstenedione reduction (EC 1.3.1.23) are catalyzed by the same catalytic site of a single protein
-
glutathione-Sepharose column chromatography and Sephacryl-S100 gel filtration
-
Ni2+-Sepharose column chromatography
-
nickel-Sepharose affinity column chromatography and Superdex 75 gel filtration
-
recombinant AKR1D1 enzyme mutant P133R from HEK-293 cells, the other expressed mutants cannot be purified
-
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity and anion exchange chromatography, and gel filtration
-
recombinant wild-type and mutant enzymes from Escherichia coli strain M15 by nickel affinity chromatography
to apparent homogeneity by affinity chromatography
-
to homogeneity by NiNTA affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, sequence comparisons, expression of wild-type and mutant enzymes in Escherichia coli strain M15
expressed in Escherichia coli
-
expressed in Escherichia coli strain C41(DE3)
-
expressed in Escherichia coli strain M15
-
expression in Escherichia coli and HEK-293 cell; recombinant expression of AKR1D1 enzyme mutants in HEK-293 cells
-
expression in human HEK-293 cells; pseudogene found
-
expression of GST-tagged enzyme in Escherichia coli
-
full-length P5betaR cDNA overexpressed in Escherichia coli as a fusion protein with MBP or His-tag
-
gene SRD5B1, DNA and amino acid sequence determination and analysis of wild-type and mutant enzymes
overproduced by the pQE expression system and transformed into Escherichia coli strain B834(DE3)
-
recombinant expression of AKR1D1 enzyme mutants in HEK-293 cells only at low levels possible and with low activities
-
Sph1/Sal1 5beta-POR fragment cloned into the pQE vector and transformed into Escherichia coli strain M15(pREP4)
-
SPHI /SALI IC5beta-POR fragment cloned into the pQE vector system and transformed into Escherichai coli strain M15(pREP4)
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
presence of chenodeoxycholic acid reduces transcript abundance in H4iiE cell and promoter activity in HepG2 cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C352G
-
site-directed mutagenesis, mutation at the substrate binding site, the mutant shows reduced activity compared to the wild-type enzyme
D181T/L182Q
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site-directed mutagenesis, mutation at motif V, the mutant shows reduced activity compared to the wild-type enzyme
F353M
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site-directed mutagenesis, mutation at the substrate binding site, the mutant shows similar activity as the wild-type enzyme
F353P
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site-directed mutagenesis, mutation at the substrate binding site, the mutant shows similar activity as the wild-type enzyme
G204N
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site-directed mutagenesis, mutation at motif IV, the mutant shows reduced activity compared to the wild-type enzyme
L182Q
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site-directed mutagenesis, mutation at motif V, the mutant shows similar activity as the wild-type enzyme
M150L
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site-directed mutagenesis, mutation at motif IV, the mutant shows similar activity as the wild-type enzyme
N205A
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site-directed mutagenesis, mutation at the substrate binding site, the mutant shows increased activity compared to the wild-type enzyme
N205M
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site-directed mutagenesis, mutation at the substrate binding site, the mutant shows increased activity compared to the wild-type enzyme
N205M/Y156V
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site-directed mutagenesis, double mutation at the substrate binding site, the mutant shows increased activity compared to the wild-type enzyme
R146T
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site-directed mutagenesis, mutation at motif IV, the mutant shows similar activity as the wild-type enzyme
S248M
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site-directed mutagenesis, mutation near the substrate binding site, inactive mutant
T65P
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site-directed mutagenesis, mutation at near motif II, the mutant shows similar activity as the wild-type enzyme
Y156V
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site-directed mutagenesis, mutation at the substrate binding site, the mutant shows increased activity compared to the wild-type enzyme
Y179A
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inactive
Y179F
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inactive
Y302F
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site-directed mutagenesis, mutation at motif IV, the mutant shows similar activity as the wild-type enzyme
E120A
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mutant is devoid of activity
R217stop
the naturally occuring mutation of gene SRD5B1 are involved in hyper-3-oxo-DELTA4 bile aciduria from primary 3-oxo-DELTA4-steroid 5beta-reductase deficiency, phenotypes, overview
V309F
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replacement of one of the residues delineating this site by a phenylalanine completely abolishes the enzyme's substrate inhibition, but the catalytic efficiency of the mutated enzyme is similar to that of the wild-type h5beta-red enzyme
Y132F
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replacement of one of the residues delineating this site by a phenylalanine completely abolishes the enzyme's substrate inhibition, but the catalytic efficiency of the mutated enzyme is similar to that of the wild-type h5beta-red enzyme
Y58F
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mutant is devoid of activity
additional information
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determination of AKR1D1 reaction mechanism by mutational analysis revealing that the 4-pro-R hydride is transferred from the re-face of the nicotinamide ring to C5 of the steroid substrate. E120, a unique substitution in the AKR catalytic tetrad, permits a deeper penetration of the steroid substrate into the active site to promote optimal reactant positioning. It participates with Y58 to create a superacidic oxyanion hole for polarization of the C3 ketone, no role for K87 in the proton relay, overview
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
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role for bile acids in inhibiting hepatic glucocorticoid clearance, of sufficient magnitude to suppress hypothalamic-pituitary-adrenal axis activity. Elevated hepatic bile acids may account for adrenal insufficiency in liver disease
additional information