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Information on EC 1.14.14.16 - steroid 21-monooxygenase and Organism(s) Bos taurus and UniProt Accession P00191
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1.14.14.16 steroid 21-monooxygenaseIUBMB Comments
A P-450 heme-thiolate protein 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. The electron donor is EC 1.6.2.4, NADPH---hemoprotein reductase.
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Word Map
- 1.14.14.16
- adrenal
- hyperplasia
- cortisol
-
virilize
- androgen
- 17-hydroxyprogesterone
- glucocorticoid
- children
- acth
-
steroidogenic
- testosterone
-
adrenocortical
-
salt-wasting
- aldosterone
- hirsutism
-
nonclassical
-
prenatal
- girl
- androstenedione
- genitalia
- steroidogenesis
-
mineralocorticoid
- hyperandrogenism
- hydrocortisone
- addison
- puberty
- 11-deoxycortisol
- dehydroepiandrosterone
-
beta-hydroxysteroid
-
11beta-hydroxylase
-
adrenarche
- p450scc
- c4a
-
endocrinologist
-
acth-stimulated
- pseudohermaphroditism
- debrisoquine
- biotechnology
-
1'-hydroxylation
- hsd3b2
-
incidentalomas
- cyp11b1
-
masculinization
- bufuralol
- medicine
- deoxycorticosterone
-
adrenocorticotropic
- 11-deoxycorticosterone
- 17alpha-hydroxylase
- dextromethorphan
-
fludrocortisone
-
hyperandrogenemia
The taxonomic range for the selected organisms is: Bos taurus
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
21-hydroxylase, cyp21a2, cyp21, cyp2d, steroid 21-hydroxylase, p450c21, progesterone 21-hydroxylase, cytochrome p450c21, p-450(c21), 21-hydroxylase cytochrome p-450, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
17alpha-hydroxyprogesterone 21-hydrolase
-
-
-
-
21-hydroxylase
-
-
-
-
21-hydroxylase cytochrome P-450
-
-
-
-
Cytochrome P-450 specific for steroid C-21 hydroxylation
-
-
-
-
cytochrome P-450-linked mixed function oxidase system
-
-
-
-
cytochrome P-450C-21
-
-
-
-
P-450(C21)
-
-
-
-
P450-C21
-
-
-
-
P450-C21B
-
-
-
-
Steroid 21-hydroxylase
-
-
-
-
steroid 21-hydroxylase system
-
-
-
-
steroid 21-hydroxylation system
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
hydroxylation
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -
SYSTEMATIC NAME
IUBMB Comments
steroid,NADPH-hemoprotein reductase:oxygen oxidoreductase (21-hydroxylating)
A P-450 heme-thiolate protein 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. The electron donor is EC 1.6.2.4, NADPH---hemoprotein reductase.
CAS REGISTRY NUMBER
COMMENTARY
9029-68-9
-
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
17alpha-hydroxyprogesterone + NADPH + O2
17,21-dihydroxyprogesterone + NADP+ + H2O
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycortisol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
progesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
progesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
-
-
-
?
(+)-benzphetamine + [reduced NADPH-cytochrome P-450 reductase] + O2
N-demethyl-benzphetamine + [oxidized NADPH-cytochrome P-450 reductase] + H2O
-
-
-
?
17alpha-hydroxyprogesterone + NADPH + H+ + O2
11-deoxycortisol + NADP+ + H2O
-
P450c21 catalyzes hydroxylation at C-21, 17alpha-hydroxyprogesterone is a better substrate for P450c21 than progesterone from the catalytic coupling with consumption of NADPH
-
-
?
a steroid + electron donor + O2
a 21-hydroxysteroid + oxidized electron donor + H2O
-
essential step in synthesis of steroid hormones by adrenal gland
-
?
progesterone + NADPH + H+ + O2
11-deoxycorticosterone + NADP+ + H2O
-
P450c21 catalyzes hydroxylation at C-21
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
two 17alpha-hydroxyprogesterone molecules are bound to the enzyme, the distal one being located at the entrance of the substrate access channel and the proximal one bound in the active site
-
-
?
17alpha-hydroxyprogesterone + NADH + O2
17,21-dihydroxyprogesterone + NAD+ + H2O
-
-
-
?
17alpha-hydroxyprogesterone + NADH + O2
17,21-dihydroxyprogesterone + NAD+ + H2O
-
-
-
?
17alpha-hydroxyprogesterone + NADH + O2
17,21-dihydroxyprogesterone + NAD+ + H2O
-
-
-
-
?
17alpha-hydroxyprogesterone + NADPH + O2
11-deoxycortisol + NADP+ + H2O
-
-
-
?
17alpha-hydroxyprogesterone + NADPH + O2
11-deoxycortisol + NADP+ + H2O
-
-
-
?
DELTA5-pregnen-3beta-ol-20-one + NADH + O2
deoxycorticosterone + NAD+ + H2O
-
-
very small yields, presumably via dehydrogenation followed by C-21 hydroxylation
?
DELTA5-pregnen-3beta-ol-20-one + NADH + O2
deoxycorticosterone + NAD+ + H2O
-
no substrate of cytochrome P-450-linked mixed function oxidase system
-
-
?
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
17alpha-hydroxyprogesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycortisol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
-
-
-
?
progesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
progesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
-
-
-
?
a steroid + electron donor + O2
a 21-hydroxysteroid + oxidized electron donor + H2O
-
essential step in synthesis of steroid hormones by adrenal gland
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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)
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
p-chloromercuribenzoate
-
complete inhibition at 1 mM, but not inhibitory at 0.1 mM
sodium o-(3-hydroxymercuri-2-methoxypropyl)carbamyl-phenoxyacetc acid (mersalyl)
-
complete inhibition at 1 mM
additional information
-
not inhibited by iodoacetate, o-iodosobenzoate, glutathione, EDTA, dipyridyl, quinacrine, ribonuclease, cytochrome c + CN-
-
additional information
-
not inhibited by superoxide dismutase, cytochrome b5, NADH-cytochrome b5 reductase
-
additional information
-
not inhibited by metal ions such as Cr3+, Fe3+, Zn2+, Pb2+, Mn2+, Co2+ at 1 mM
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0108
17alpha-hydroxyprogesterone
37°C, pH 7.4, presence of 0.002% Cymal 5
additional information
additional information
Pre-steady-state and steady-state binding kinetics with 17alpha-hydroxyprogesterone, stopped-flow spectroscopic measurements, overview
-
additional information
additional information
-
Pre-steady-state and steady-state binding kinetics with 17alpha-hydroxyprogesterone, stopped-flow spectroscopic measurements, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.22
17alpha-hydroxyprogesterone
pH 7.4, 37°C, CYP21 wild-type and mutant T241R/L442A
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0195
-
cytochrome P-450-linked mixed function oxidase system from adrenal gland, 21-hydroxylation of 17alpha-hydroxyprogesterone
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
deficiency of this enzyme is involved in about 95% of cases of human congenital adrenal hyperplasia
additional information
additional information
the key substrate recognition residues are not only around the heme but also along the substrate access channel, structure-function analysis, overview
additional information
-
the key substrate recognition residues are not only around the heme but also along the substrate access channel, structure-function analysis, overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). CP21A_BOVIN
496
1
56077
Swiss-Prot
Secretory Pathway (Reliability: 1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
12400
gel filtration in presence of 0.12% Cymal 5, protein elutes as monomer surrounded by a micelle of detergent
167000
gel filtration in presence of 0.056% Cymal 6, protein elutes as monomer surrounded by a micelle of detergent
80700
gel filtration in presence of 1% cholate, protein elutes as monomer surrounded by a micelle of cholate
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
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
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant His-tagged CYP21A2 mutant T241R/L442A in complex with substrate 17-hydroxyprogesterone, hanging drop vapor diffusion method, 0.2 mM protein in 50 mM potassium phosphate, pH 7.4, 20% glycerol, 0.1 mM DTT, 0.1 mM EDTA, 0.25% Cymal 5, and 50 mM NaCl, is mixed with 0.4 mM, containing 2% v/v C2H5OH, and 5-15% w/v PEG 3350, 0.5 M ammonium sulfate, and 0.1 M HEPES, pH 7.0, 20°C, few days, X-ray diffraction structure determination and analysis at 3.0 A resolution
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
T241R
site-directed mutagenesis, the mutant shows improved solubility properties compared to the wild-type enzyme
T241R/L442A
site-directed mutagenesis, the mutant shows greatly improved solubility properties compared to the wild-type enzyme
additional information
additional information
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
additional information
-
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
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70°C, 0.2 M potassium phospate, pH 7.4, 20% glycerol, 0.1 mM EDTA, 3 months without loss of activity
-
0°C, 50 mM Tris, pH 7.2, 20% glycerol, 0.15% emulgen, 0.1 mM dithiothreitol, 0.1 mM EDTA, several weeks spectroscopically stable
-
25°C, 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
LITERATURE
recombinant His-tagged wild-type enzyme and mutants from Escherichia coli by nickel affinity and anion exchange chromatography, followed by gel filtration
enzyme system consisting of cytochrome P-450S21 and NADPH-cytochrome P-450 reductase
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene CYP21A2 or C3B21RA, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged wild-type enzyme and mutants in Escherichia coli
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ryan, K.J.; Engel, L.L.
Hydroxylation of steroids at carbon 21
J. Biol. Chem.
225
103-114
1957
Bos taurus
Hiwatashi, A.; Ichikawa, Y.
Purification and reconstitution of the steroid 21-hydroxylase system (cytochrome P-450-linked mixed function oxidase system) of bovine adrenocortical microsomes
Biochim. Biophys. Acta
664
33-48
1981
Bos taurus
Greenfield, N.; Ponticorvo, L.; Chasalow, F.; Lieberman, S.
Activation and Inhibition of the adrenal steroid 21-hydroxylation system by cytosolic constituents: influence of glutathione, glutathione reductase, and ascorbate
Arch. Biochem. Biophys.
200
232-244
1980
Bos taurus
Narasimhulu, S.; Eddy, C.R.
Adrenal microsomal hydroxylating system: purification and substrate binduing properties of cytochrome P-450C-21
Biochemistry
24
4287-4294
1985
Bos taurus
Belanger, A.; Tremblay, Y.; Vallee, M.; Provencher, P.H.; Perron, S.
Regulation of 21 hydroxylase activity by steroids
Endocr. Res.
21
329-41
1995
Bos taurus
Kominami, S.; Ochi, H.; Kobayashi, Y.; Takemori, S.
Studies on the steroid hydroxylation system in adrenal cortex microsomes
J. Biol. Chem.
255
3386-3394
1980
Bos taurus
Bumpus, J.A.; Dus, K.
Bovine adrenocortical microsomal hemeproteins P-45017alpha and P-450C-21: Isolation, partial characterization, and comparison to P-450SCC
J. Biol. Chem.
257
12696-12704
1982
Bos taurus
Arase, M.; Waterman, M.R.; Kagawa, N.
Purification and characterization of bovine steroid 21-hydroxylase (P450c21) efficiently expressed in Escherichia coli
Biochem. Biophys. Res. Commun.
344
400-405
2006
Bos taurus (P00191), Bos taurus
Tosha, T.; Kagawa, N.; Arase, M.; Waterman, M.; Kitagawa, T.
Interaction between substrate and oxygen ligand responsible for effective O-O bond cleavage in bovine cytochrome P450 steroid 21-hydroxylase proved by Raman spectroscopy
J. Biol. Chem.
283
3708-3717
2008
Bos taurus
Zhao, B.; Lei, L.; Kagawa, N.; Sundaramoorthy, M.; Banerjee, S.; Nagy, L.D.; Guengerich, F.P.; Waterman, M.R.
Three-dimensional structure of steroid 21-hydroxylase (cytochrome P450 21A2) with two substrates reveals locations of disease-associated variants
J. Biol. Chem.
287
10613-10622
2012
Bos taurus (P00191), Bos taurus, Homo sapiens (P08686)
Pallan, P.S.; Wang, C.; Lei, L.; Yoshimoto, F.K.; Auchus, R.J.; Waterman, M.R.; Guengerich, F.P.; Egli, M.
Human cytochrome P450 21A2, the major steroid 21-hydroxylase: structure of the enzyme-progesterone substrate complex and rate-limiting C-H bond cleavage
J. Biol. Chem.
290
13128-13143
2015
Bos taurus (P00191), Bos taurus, Homo sapiens (P08686), Homo sapiens
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