Information on EC 1.14.99.10 - steroid 21-monooxygenase

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The expected taxonomic range for this enzyme is: Euteleostomi

EC NUMBER
COMMENTARY
1.14.99.10
-
RECOMMENDED NAME
GeneOntology No.
steroid 21-monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
a C21 steroid + [reduced NADPH-hemoprotein reductase] + O2 = a 21-hydroxy-C21-steroid + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydroxylation
-
-
-
-
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Metabolic pathways
-
Steroid hormone biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
steroid,NADPH-hemoprotein reductase:oxygen oxidoreductase (21-hydroxylating)
Requires NADPH and EC 1.6.2.4, NADPH---hemoprotein reductase. A heme-thiolate protein (P-450) enzyme responsible for the conversion of progesterone and 17alpha-hydroxyprogesterone to their respective 21-hydroxylated derivatives, 11-deoxycorticosterone and 11-deoxycortisol. Involved in the biosynthesis of the hormones aldosterone and cortisol.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
17alpha-hydroxyprogesterone 21-hydrolase
-
-
-
-
21-hydroxylase
-
-
-
-
21-hydroxylase
P08686
-
21-hydroxylase cytochrome P-450
-
-
-
-
21OH
-
-
21OH
P08686
-
CYP21
-
-
CYP21
P08686
-
CYP21A2
-
-
Cytochrome P-450 specific for steroid C-21 hydroxylation
-
-
-
-
cytochrome P-450-linked mixed function oxidase system
-
-
-
-
cytochrome P-450C-21
-
-
-
-
cytochrome P450 steroid 21-hydroxylase
-
-
EC 1.14.1.8
-
-
formerly
-
EC 1.99.1.11
-
-
formerly
-
P-450(C21)
-
-
-
-
P450 oxidoreductase-21-hydroxylase
-
-
P450-C21
-
-
-
-
P450-C21B
-
-
-
-
P450c21
-
-
P450c21
P00191
-
P450c21
-
-
Steroid 21-hydroxylase
-
-
-
-
Steroid 21-hydroxylase
-
-
steroid 21-hydroxylase system
-
-
-
-
steroid 21-hydroxylation system
-
-
-
-
steroid cytochrome P450 21-hydroxylase
-
-
steroid cytochrome P450 21-hydroxylase
P08686
-
CAS REGISTRY NUMBER
COMMENTARY
9029-68-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
expressed in Escherichia coli
SwissProt
Manually annotated by BRENDA team
expression in human embryonic kidney 293 cells
-
-
Manually annotated by BRENDA team
expression in COS-1 cells
-
-
Manually annotated by BRENDA team
gene CYP21A2
-
-
Manually annotated by BRENDA team
Italian patients with congenital adrenal hyperplasia
-
-
Manually annotated by BRENDA team
Middle European patients with congenital adrenal hyperplasia
-
-
Manually annotated by BRENDA team
rainbow trout
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
steroid 21-hydroxylase deficiency accounts for more than 90% of congenital adrenal hyperplasia
malfunction
-
the naturally occuring 21-hydroxylase mutation H365Y/R356W is involved in congenital adrenal hyperplasia, an autosomal recessive The CYP21A2 H365Y mutant exhibits minimal 21-hydroxylase activity to convert 17-hydroxyprogesterone to 11-deoxycortisol or progesterone to 11-deoxycorticosterone. The H365Y mutant protein may be unstable and/or subject to a more rapid degradation by the human proteosome as well as catalytically inefficient. The double mutant genotype with a severe mutation on each allele is compatible with the clinical presentation
additional information
-
In autoimmune adrenal deficiency, autoantibodies target the 21-hydroxylase protein, 21-hydroxylase-specific T cells are CD8+ T cells. A T-cell epitope mapping study using 49 overlapping 20mer peptides covering the 21OH sequence in patients with isolated Addison's disease, Autoimmune Polyendocrine Syndrome 1 and 2 for determination of the epitopes targeted in autoimmune adrenal deficiency, detailed overview
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(+)-benzphetamine + NADH + O2
N-demethyl-benzphetamine + NAD+ + H2O
show the reaction diagram
-
-
-
?
11,17-dihydroxyprogesterone + NADH + O2
cortisol + NAD+ + H2O
show the reaction diagram
-
-
-
?
11beta-hydroxyprogesterone + NADH + O2
corticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
17-hydroxy-progesterone + NADH + O2
17-hydroxy-11-deoxy-corticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
17-hydroxy-progesterone + NADH + O2
17-hydroxy-11-deoxy-corticosterone + NAD+ + H2O
show the reaction diagram
-
isozyme CYP2D
-
-
?
17-hydroxyprogesterone + NADPH + H+ + O2
11-deoxycortisol + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
17-hydroxyprogesterone + NADPH + H+ + O2
11-deoxycortisol + NADP+ + H2O
show the reaction diagram
-
the redox cofactor provides the electrons required for a P450-dependent hydroxylation. Maximal CYP 21 activity is observed in presence of low substrate concentrations of 0.2 mM. Higher substrate concentrations reduce the activity and inhibit the enzyme
-
-
?
17-hydroxyprogesterone + NADPH + O2
corticosteroids + NADP+ + H2O
show the reaction diagram
-
-
-
?
17alpha-hydroxy-progesterone + NADH + O2
17alpha-hydroxy-11-deoxy-corticosterone + NAD+ + H2O
show the reaction diagram
-
preferred substrate
-
-
?
17alpha-hydroxyprogesterone + AH2 + O2
11-deoxycortisol + A + H2O
show the reaction diagram
-
-
-
-
?
17alpha-hydroxyprogesterone + NADH + O2
17,21-dihydroxyprogesterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
17alpha-hydroxyprogesterone + NADH + O2
17,21-dihydroxyprogesterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
17alpha-hydroxyprogesterone + NADH + O2
17,21-dihydroxyprogesterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
17alpha-hydroxyprogesterone + NADH + O2
17,21-dihydroxyprogesterone + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
17alpha-hydroxyprogesterone + NADH + O2
17,21-dihydroxyprogesterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
17alpha-hydroxyprogesterone + NADPH + H+ + O2
11-deoxycortisol + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
17alpha-hydroxyprogesterone + NADPH + H+ + O2
11-deoxycortisol + NADP+ + H2O
show the reaction diagram
-
P450c21 catalyzes hydroxylation at C-21, 17alpha-hydroxyprogesterone is a better substrate for P450c21 than progesterone from the catalytic coupling with consumption of NADPH
-
-
?
17alpha-hydroxyprogesterone + NADPH + O2
11-deoxycortisol + NADP+ + H2O
show the reaction diagram
-
-
-
?
17alpha-hydroxyprogesterone + NADPH + O2
11-deoxycortisol + NADP+ + H2O
show the reaction diagram
-
-
-
?
17alpha-hydroxyprogesterone + NADPH + O2
17,21-dihydroxyprogesterone + NADP+ + H2O
show the reaction diagram
P00191
-
-
-
?
a steroid + electron donor + O2
a 21-hydroxysteroid + oxidized electron donor + H2O
show the reaction diagram
-
essential step in synthesis of steroid hormones by adrenal gland
-
-
allopregnanolone + NADH + O2
21-hydroxy-allopregnanolone + NAD+ + H2O
show the reaction diagram
-
isozyme CYP2D4, CYP2D6
-
-
?
DELTA5-pregnen-3beta,17alpha-diol-20-one + NADH + O2
? + NAD+ + H2O
show the reaction diagram
-
-
-
-
-
DELTA5-pregnen-3beta-ol-20-one + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
very small yields, presumably via dehydrogenation followed by C-21 hydroxylation
?
progesterone + AH2 + O2
deoxycorticosterone + A + H2O
show the reaction diagram
-
-
-
-
?
progesterone + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
progesterone + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
progesterone + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
-
-
progesterone + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
progesterone + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
progesterone + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
progesterone + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
progesterone + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
isozyme CYP2D
-
-
?
progesterone + NADH + O2
11-deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
progesterone + NADPH + H+ + O2
11-deoxycorticosterone + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
progesterone + NADPH + H+ + O2
11-deoxycorticosterone + NADP+ + H2O
show the reaction diagram
-
P450c21 catalyzes hydroxylation at C-21
-
-
?
progesterone + NADPH + H+ + O2
deoxycorticosterone + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
progesterone + NADPH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
progesterone + NADPH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
-
-
?
progesterone + NADPH + O2
deoxycorticosterone + NADP+ + H2O
show the reaction diagram
P00191
-
-
-
?
DELTA5-pregnen-3beta-ol-20-one + NADH + O2
deoxycorticosterone + NAD+ + H2O
show the reaction diagram
-
no substrate of cytochrome P-450-linked mixed function oxidase system
-
-
-
additional information
?
-
-
no substrate: 17alpha-hydroxy-progesterone
-
-
-
additional information
?
-
-
enzyme deficiency, i.e. 21-OHD, causes congenital adrenal hyperplasia in pubertal subjects
-
-
-
additional information
?
-
-
CYP21 performs the regio- and stereo-selective 21-hydroxylation of 17-alpha-hydroxyprogesterone when recombinantly expressed in Schizosaccharomyces pombe, reaction method optimization, overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
17-hydroxyprogesterone + NADPH + H+ + O2
11-deoxycortisol + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
17-hydroxyprogesterone + NADPH + O2
corticosteroids + NADP+ + H2O
show the reaction diagram
-
-
-
?
progesterone + NADPH + H+ + O2
11-deoxycorticosterone + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
a steroid + electron donor + O2
a 21-hydroxysteroid + oxidized electron donor + H2O
show the reaction diagram
-
essential step in synthesis of steroid hormones by adrenal gland
-
-
additional information
?
-
-
enzyme deficiency, i.e. 21-OHD, causes congenital adrenal hyperplasia in pubertal subjects
-
-
-
additional information
?
-
-
CYP21 performs the regio- and stereo-selective 21-hydroxylation of 17-alpha-hydroxyprogesterone when recombinantly expressed in Schizosaccharomyces pombe
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
cytochrome P450
-
-
-
cytochrome P450
-
-
-
cytochrome P450
-
-
-
cytochrome P450
-
Cyp21 is a cytochrome P450 monooxygenase
-
cytochrome P450S21
-
steroid 21-hydroxylase system consists of cytochrome P-450S21, NADPH-cytochrome P-450 reductase (EC 1.6.2.4) and steroid 21-monooxygenase (EC 1.14.99.10)
-
heme
-
0.8 mol/mol peptide
NADH
-
NADH is 50% as effective as NADPH
NADPH
-
the redox cofactor provides the electrons required for a P450-dependent hydroxylation
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Fe2+
-
a cytochrome P450 monooxygenase
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
17alpha-progesterone
-
inhibits above 0.2mM
-
antibody to cytochrome P-450BPA
-
-
-
antibody to NADPH-cytochrome P-450 reductase
-
-
-
antimycin A
-
15% inhibition at 1mg per l
ascorbate
-
10% inhibition at 10 mM
ascorbate
-
-
azide
-
9% inhibition at 1 mM
carbon monoxide
-
dark 65% inhibition at 90%, light 57% inhibition at 90%
carbon monoxide
-
-
CuSO4
-
50% inhibition at 1 mM
cyanide
-
14% inhibition at 1 mM
cytochrome c
-
complete inhibition at 0.1 mM, 0.003 mM
diethylaminoethyldiphenylpropylacetic acid SKF 525 A
-
9% inhibition at 1 mM
fluoxetine
-
50% inhibition at 0.002 mM, substrate allopregnanolone
p-chloromercuribenzoate
-
complete inhibition at 1 mM, but not inhibitory at 0.1 mM
Phenylisocyanide
-
inhibition at 0.5 mM
resveratrol
-
decreases adrenal enzyme expression in vivo and in cell culture. Corticosterone production is inhibited 47% by 0.05 mM resveratrol in vitro and 20% ex vivo, while progesterone production is elevated to 400% of control in vitro
RU38486
-
-
sodium o-(3-hydroxymercuri-2-methoxypropyl)carbamyl-phenoxyacetc acid (mersalyl)
-
complete inhibition at 1 mM
HgCl2
-
complete inhibition at 0.1 mM
additional information
-
not inhibited by catalase; not inhibited by iodoacetate, o-iodosobenzoate, glutathione, EDTA, dipyridyl, quinacrine, ribonuclease, cytochrome c + CN-; not inhibited by metal ions such as Cr3+, Fe3+, Zn2+, Pb2+, Mn2+, Co2+ at 1 mM
-
additional information
-
not inhibited by catalase; not inhibited by superoxide dismutase, cytochrome b5, NADH-cytochrome b5 reductase
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
-
Bovine serum albumin
-
-
-
Cymal 5
P00191
optimal concentration is 0.002%, decrease in activity above 0.05%
dithiothreitol
-
-
Emulgen 911
-
-
-
Emulgen 913
-
maximum with 0.008% v/v
L-cysteine
-
-
L-cystine
-
-
lysophosphatidylcholine
-
-
Sodium cholate
-
-
Sodium deoxycholate
-
-
Triton X-100
-
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2.3
-
17-hydroxy-progesterone
-
37C, mutant P482S
8.5
-
17-hydroxy-progesterone
-
37C, wild-type
9.2
-
17-hydroxy-progesterone
-
37C, mutant A15T
0.0038
-
17-hydroxyprogesterone
-
head kidney microsomes
0.0055
-
17-hydroxyprogesterone
-
-
0.031
-
17-hydroxyprogesterone
-
liver microsomes
0.00135
-
17alpha-hydroxyprogesterone
-
mutant enzyme D322G, in 50 mM potassium phosphate buffer (pH 7.4), at 37C
0.00138
-
17alpha-hydroxyprogesterone
-
wild type enzyme, in 50 mM potassium phosphate buffer (pH 7.4), at 37C
0.0016
-
17alpha-hydroxyprogesterone
-
wild type enzyme
0.00181
-
17alpha-hydroxyprogesterone
-
mutant enzyme A265V, in 50 mM potassium phosphate buffer (pH 7.4), at 37C
0.002
-
17alpha-hydroxyprogesterone
-
mutant enzyme P453S
0.0026
-
17alpha-hydroxyprogesterone
-
mutant enzyme K121Q
0.00072
-
progesterone
-
mutant enzyme D322G, in 50 mM potassium phosphate buffer (pH 7.4), at 37C; wild type enzyme, in 50 mM potassium phosphate buffer (pH 7.4), at 37C
0.00079
-
progesterone
-
mutant enzyme A265V, in 50 mM potassium phosphate buffer (pH 7.4), at 37C
0.0015
-
progesterone
-
mutant enzyme P453S; wild type enzyme
0.0024
-
progesterone
-
mutant enzyme K121Q
0.0055
-
progesterone
-
-
0.0129
-
progesterone
P00191
37C, pH 7.4, presence of 0.002% Cymal 5
3
-
progesterone
-
37C, mutant P482S
6.4
-
progesterone
-
37C, wild-type
9.2
-
progesterone
-
37C, mutant A15T
0.0108
-
17alpha-hydroxyprogesterone
P00191
37C, pH 7.4, presence of 0.002% Cymal 5
additional information
-
additional information
-
reaction kinetic, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.322
-
17a-hydroxyprogesterone
-
-
0.172
-
progesterone
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.00027
-
-
37C, mutant P482S, substrate 17-hydroxy-progesterone
0.00038
-
-
37C, mutant P482S, substrate progesterone
0.00117
-
-
37C, wild-type, substrate progesterone
0.00122
-
-
37C, wild-type, substrate 17-hydroxy-progesterone
0.00129
-
-
37C, mutant A15T, substrate 17-hydroxy-progesterone
0.00148
-
-
37C, mutant A15T, substrate progesterone
0.003
-
-
21-hydroxylation of progesterone at 26C
0.0038
-
-
21-hydroxylation of 17alpha-hydroxyprogesterone at 26C
0.0052
-
-
N-demethylation of (+)-benzphetamine
0.0077
-
-
21-hydroxylation of progesterone
0.0144
-
-
21-hydroxylation of progesterone
0.0195
-
-
cytochrome P-450-linked mixed function oxidase system from adrenal gland, 21-hydroxylation of 17alpha-hydroxyprogesterone
0.0452
-
-
21-hydroxylation of 17alpha-hydroxyprogesterone
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
7
-
-
7.4
-
-
-
7.5
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
8
-
about 50% of activity maximum at pH 5.5 and 8.0
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
sole source of enzyme expression
Manually annotated by BRENDA team
-
anterior qurter of head kidney
Manually annotated by BRENDA team
additional information
-
no activity in: gill, heart, liver, intestine, kidney, immature gonad, skeletal muscle
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
microsomal steroid 21-hydroxylation is not catalyzed by P450c21, but by CYP2D isoforms
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
12400
-
P00191
gel filtration in presence of 0.12% Cymal 5, protein elutes as monomer surrounded by a micelle of detergent
47000
-
-
SDS-PAGE
47500
-
-
SDS-PAGE
48870
-
-
calculation form amino acid composition
52000
-
-
SDS-PAGE
54000
-
-
SDS-PAGE
54000
-
-
SDS-PAGE, chromatography on Sephadex
80700
-
P00191
gel filtration in presence of 1% cholate, protein elutes as monomer surrounded by a micelle of cholate
167000
-
P00191
gel filtration in presence of 0.056% Cymal 6, protein elutes as monomer surrounded by a micelle of detergent
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
monomer
P00191
1 * 53000, calculated
additional information
-
steroid 21-hydroxylase system consists of cytochrome P-450S21, NADPH-cytochrome P-450 reductase, EC 1.6.2.4, and steroid 21-monooxygenase, EC 1.14.99.10
additional information
-
monomer-dimer equilibrium
additional information
-
amino acid analysis, N-terminal sequence, comparison with other cytochromes P-450
additional information
-
amino acid analysis
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
3.6% carbohydrate
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-70C, 0.2 M potassium phospate, pH 7.4, 20% glycerol, 0.1 mM EDTA, 3 months without loss of activity
-
0C, 50 mM Tris, pH 7.2, 20% glycerol, 0.15% emulgen, 0.1 mM dithiothreitol, 0.1 mM EDTA, several weeks spectroscopically stable
-
25C, 50 mM Tris, pH 7.2, 20% glycerol, 0.15% emulgen, 0.1 mM dithiothreitol, 0.1 mM EDTA, several hours spectroscopically stable, without Emulgen 50% precipitation after 30 min
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
enzyme system consisting of cytochrome P-450S21 and NADPH-cytochrome P-450 reductase
-
HisTrap HP column chromatography (using Ni2+ ions immobilized onto a metal-chelating column)
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli
-
expressed in COS-1 cells
-
expressed in Sf9 insect cell microsomes
-
expressed in Spodoptera frugiperda insect cell microsomes using a baculovirus expression system
-
gene CYP11B!, DNA and amino acid sequence determination and analysis, genotyping
-
gene CYP21, expression in Schizosaccharomyces pombe strain CAD18, method optimization and evaluation, and quantification of the intracellular redox cofactor pool in transformed cells, overview
-
gene CYP21A2, DNA and amino acid sequence determination and analysis, genotyping. Recombinant expression of mutant H365Y/R356W in HEK-293T cells
-
mutant enzymes are expressed in COS-7 cells
-
mutant enzymes are expressed in Saccharomyces cerevisiae strain W303B
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A15T
-
natural mutation found in patients with classical congenital adrenal hyperplasia, no significant difference in activity compared to wild-type
A265V
-
the mutant enzyme activity is similar to wild type
D322G
-
the mutation impacts significantly on enzyme function and exerts activity compatible with non-classical congenital adrenal hyperplasia, has about 27% activity for the conversion of progesterone to 11-deoxycorticosterone and 18% activity for the conversion of 17alpha-hydroxyprogesterone to 11-deoxycortisol compared to wild type activity
E351K
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rare missense mutation located in the ERR triad and found in a patient with virilizing congenital hyperplasia. Residual activity is about 1% of wild-type for both 17-hydroxyprogesterone and progesterone
H365Y/R356W
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a naturally occuring 21-hydroxylase mutation in the CYP21A2 gene, that is involved in congenital adrenal hyperplasia, an autosomal recessive disorder, phenotype, overview. The H365Y enzyme is produced in more variable amounts than wild type
I171N
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mutation identified in Italian patient with congenital adrenal hyperplasia, less than 1% of wild-type enzyme activity
I172N
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naturally occuring mutant, 0-2% of wild-type activity, dominant negative effect over wild-type with 11% decrease in activity
I172N
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the mutation is associated with congenital adrenal hyperplasia
I236N/V237E/M239K
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naturally occuring mutant, no enzymic activity, dominant negative effect over wild-type with 35% decrease in activity
K122Q
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missense mutation causing nonclassical 21-hydroxylase deficiency, shows reduced activity of 14% for the conversion of 17alpha-hydroxyprogesterone and 19% for the conversion of progesterone compared to wild type
L236N/V237E/M239K
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the mutation is associated with congenital adrenal hyperplasia
P30L
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missense mutation associated with congenital adrenal hyperplasia
P30L
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the mutation is associated with congenital adrenal hyperplasia
P453S
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the mutant shows a reduced activity of 36% of wild type for the conversion of 17alpha-hydroxyprogesterone and 44% for the conversion of progesterone
P482S
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natural mutation found in patients with nonclassical congenital adrenal hyperplasia, precocious pubarche, menstrual irregularities or hypertrichosis, about 70% of activity compared to wild-type
Q318X
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the mutation is associated with congenital adrenal hyperplasia
R341P
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mutation identified in Italian patient with congenital adrenal hyperplasia, less than 1% of wild-type enzyme activity
R356W
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naturally occuring mutant, no enzymic activity, no dominant negative effect on wild-type
R356W
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the mutation is associated with congenital adrenal hyperplasia
R426H
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mutation identified in Italian patient with congenital adrenal hyperplasia, less than 1% of wild-type enzyme activity
R483Q
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the mutant enzyme activities in the conversion of progesterone to deoxycorticosterone and 17alpha-hydroxyprogesterone to 11-deoxycortisol are measured as 2.0 and 1.89% of the wild type, respectively
V281L
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naturally occuring mutant, 50% of wild-type activity, dominant negative effect over wild-type with 30% decrease in activity
V281L
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the mutation is associated with congenital adrenal hyperplasia
W302S
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the mutation impacts significantly on enzyme function and exerts activity compatible with simple virilizing congenital adrenal hyperplasia, has residual enzyme activity of about 3% compared to wild type activity for both, the conversion of progesterone to 11-deoxycorticosterone and the conversion of 17alpha-hydroxyprogesterone to 11-deoxycortisol
additional information
P00191
replacement of N-terminal membrane anchor and basic regions by the basic regions of CYP2C3 for efficient expression and purification. N-terminal membrane anchor and sequence of the basic region do not significantly affect either substrate-specificity or 21-hydroxylase activites
L446P
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mutation identified in Italian patient with congenital adrenal hyperplasia, less than 1% of wild-type enzyme activity
additional information
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enzyme deficiency, i.e. 21-OHD, causes congenital adrenal hyperplasia, growth phenotypes of pubertal humans with salt wasting, simple virilizing, or non-classical 21-OHD, respectively, overview
additional information
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deletions/conversions involving the promoter region of the CYP21A2 gene (IVS2-12C/A>G, F306-L307insT) are associated with congenital adrenal hyperplasia
additional information
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an insertion (duplication) of 9-bp in exon 2 results in addition of three valine residues at codon 71 of the P450c21 protein lowering the structural stability of P450c21 thereby leading to the probable loss of its function
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
biotechnology
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the CYP21 expression model system using resting Schizosaccharomyces pombe cells can be used for biotransformations
medicine
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dominant negative effect of mutant enzymes involved in congenital adrenal hyperplasia on wild-type alleles
medicine
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study on the effects of enzyme-specific monoclonal antibodies on enzyme activity by comparative structural modelling. Antibodies with epitopes located distant from the enzyme active sites have no effect on activity. An antibody with epitope close to the redox binding protein binding site results in almost 50% decrease in activity
medicine
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molecular analysis of enzyme gene from Middle European patients with congenital adrenal hyperplasia. CYP21 enzyme gene deletion and In2 and I172N mutation account for 72.7% of the affected alleles in the whole study group. With exception of I172N and P30L mutations, a good genotype-phenotype correlation is observed. Using high-resulotion genotyping, the causative mutation could be identified in 341 out of 348 patients