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
Steroid hormone biosynthesis
-
Metabolic pathways
-
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
-
C3B21RA protein
P00191
-
CYP21
P08686
-
CYP21A2
P00191
-
CYP21A2
P08686
-
Cytochrome P-450 specific for steroid C-21 hydroxylation
-
-
-
-
cytochrome P-450-linked mixed function oxidase system
-
-
-
-
cytochrome P-450C-21
-
-
-
-
cytochrome p450 21A2
P00191
-
cytochrome p450 21A2
P08686
-
cytochrome P450 steroid 21-hydroxylase
-
-
cytochrome P450c21
P08686
-
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
-
Progesterone 21-hydroxylase
P08686
-
Steroid 21-hydroxylase
-
-
-
-
Steroid 21-hydroxylase
P00191
-
Steroid 21-hydroxylase
-
-
Steroid 21-hydroxylase
P08686
-
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
gene CYP21A2 or C3B21RA
SwissProt
Manually annotated by BRENDA team
expression in human embryonic kidney 293 cells
-
-
Manually annotated by BRENDA team
; gene CYP21A2
-
-
Manually annotated by BRENDA team
expression in COS-1 cells
-
-
Manually annotated by BRENDA team
gene CYP21A2
UniProt
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
-
mutations in the 21-hydroxylase CYP21A2 gene, e.g. H365Y, cause the autosomal recessive disorder congenital adrenal hyperplasia, phenotype, overview; 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
malfunction
-
steroid 21-hydroxylase deficiency accounts for about 95% of individuals with congenital adrenal hyperplasia, a common autosomal recessive metabolic disorder of adrenal steroidogenesis, mutations on CYP21A2 activity lead to impairment of the synthesis of cortisol and aldosterone and the excessive production of androgens
malfunction
-
deficiency of CYP21A2 leading to congenital adrenal hyperplasia, is the most common autosomal recessive disease in human beings
physiological function
-
CYP21 catalyses reactions of progesterone and 17alpha-hydroxyprogesterone conversion to 11-deoxycorticosterone and 11-deoxycortisol, respectively, providing an important step in aldosterone and cortisol biosynthesis. Direct molecular interactions, with electrostatic interactions playing a crucial role, between steroidogenic enzymes CYP17, EC 1.14.99.9, and CYP21 that are localized in endoplasmic reticulum membranes of adrenal cortex and involved in biosynthesis of corticosteroid hormones. The interaction in vitro reduces the catalytic activities of both enzymes at high ionic strength, i.e. 300 mM NaCl, while it increases activity at low ionic strength, i.e. 100 mM NaCl, overview
physiological function
-
CYP21A2 activities are required for cortisol and aldosterone biosynthesis, and involve the formation of energetically disfavored primary carbon radicals
physiological function
-
human steroidogenic cytochrome P450 CYP21A2 is required for the biosynthesis of androgens, glucocorticoids, and mineralocorticoids
malfunction
P08686
mutations in the cytochrome p450 (CYP)21A2 gene, which encodes the enzyme steroid 21-hydroxylase, cause the majority of cases in congenital adrenal hyperplasia, an autosomal recessive disorder. The mutations can be associated either with severe salt-wasting or simple virilizing phenotypes or with milder nonclassical phenotypes, structure-phenotype correlations, overview. Mutations that affect membrane anchoring, disrupt heme and/or substrate binding, or impair stability of CYP21A2 cause complete loss of function and salt-wasting disease, while mutations altering the transmembrane region or conserved hydrophobic patches cause up to a 98% reduction in enzyme activity and simple virilizing disease
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
additional information
-
residues L107, L109, V470, I471, and V359 contribute to the CYP21A2 substate-binding pocket. Progesterone binds to CYP21A2 in a fundamentally different orientation than to CYP17A1, EC 1.14.99.9, expansion of the CYP21A2 substrate-binding pocket allows additional substrate trajectories and metabolic switching. CYP21A2 structure modelling, overview
additional information
P00191
the key substrate recognition residues are not only around the heme but also along the substrate access channel, structure-function analysis, overview
additional information
P08686
structure homology modelling using bovine CYP21A2 crystal structure as template. Close contact between the carbon of P30 in the N-terminal loop and the side chain of Y376 within the beta5-beta6 hairpin loop is critical for protein folding
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
-
-
-
?
17,17-dimethyl-18-norandrosta-1,4,13-trien-3-one + [reduced NADPH-hemoprotein reductase] + O2
16beta-hydroxy-17,17-dimethyl-18-norandrosta-1,4,13-trien-3-one + 16alpha-hydroxy-17,17-dimethyl-18-norandrosta-1,4,13-trien-3-one + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
i.e. metandienone
product identification by GC-EI-MS analysis
-
?
17,17-dimethyl-18-norandrosta-1,4,13-trien-3-one + [reduced NADPH-hemoprotein reductase] + O2
20beta-hydroxy-17,17-dimethyl-18-norandrosta-1,4,13-trien-3-one + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
i.e. metandienone
product identification by GC-EI-MS analysis
-
?
17-fluoro-progesterone + [reduced NADPH-P450 reductase] + O2
17-fluoro-11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + 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
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycortisol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycortisol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
preferred substrate. The 17alpha-hydroxyprogesterone metabolites formed by mutants of V470 and I471 include a second, unidentified product. Both mutants V359A and V359G also metabolize 17alpha-hydroxyprogesterone to 11-deoxycortisol and this second product, and for mutation V359G, the unidentified compound is the major product. The second compound is not androstenedione and is neither the 16alpha- nor 6beta-hydroxylation products (pregn-4-ene-16alpha,17alpha-diol-3,20-dione and pregn-4-ene-6beta,17alpha-diol-3,20-dione, respectively)
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
-
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
P00191
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
-
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
P00191
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
-
-
?
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 + NADP+ + H2O
show the reaction diagram
-
-
-
?
progesterone + NADPH + O2
deoxycorticosterone + NADP+ + H2O
show the reaction diagram
-
-
-
?
progesterone + NADPH + O2
deoxycorticosterone + NADP+ + H2O
show the reaction diagram
P00191
-
-
-
?
progesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
progesterone + [reduced NADPH-hemoprotein reductase] + O2
16alpha-hydroxy-progesterone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
mutant V359A produces an additional metabolite, 16alpha-hydroxyprogesterone of progesterone, very low activity with the wild-type enzyme
product identification by TLC
-
?
progesterone + [reduced NADPH-hemoprotein reductase] + O2
16alpha-hydroxyprogesterone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
mutants V359A and V359G both produce an additional metabolite, 16alpha-hydroxyprogesterone
product identification by TLC
-
?
progesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
-
-
-
-
?
progesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
P00191
-
-
-
?
progesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
-
-
-
-
?
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
-
-
-
additional information
?
-
-
metandienone hydroxylation activities and metabolism in vivo, product analysis, overview
-
-
-
additional information
?
-
-
17alpha-hydroxyprogesterone is hydroxylated 28times faster than progesterone by the wild-type enzyme. No activity of the wild-type enzyme with pregnenolone, 17alpha-hydroxypregnenolone, and 5alpha-pregnane-3alpha-ol-20-one (allopregnanolone)
-
-
-
additional information
?
-
-
21-trifluorosteroids mht by a substrates for CYP21A2, substrate specificity analysis of CP21A2 with diverse halogenated steroid substrates, e.g. introducing one or more halogen atom to the 17- and 21-positions of progesterone and pregnenolone, 21,21,21-tribromoprogesterone and 21,21,21-trichloroprogesterone, overview
-
-
-
additional information
?
-
-
human CYP21A2 also has progesterone 16alpha-hydroxylase activity. Product analysis by LC-MS/MS
-
-
-
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
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycortisol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
-
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
P00191
-
-
-
?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
-
-
-
-
?
progesterone + NADPH + H+ + O2
11-deoxycorticosterone + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
progesterone + [reduced NADPH-hemoprotein reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
progesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
-
-
-
-
?
progesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
show the reaction diagram
P00191
-
-
-
?
progesterone + [reduced NADPH-P450 reductase] + O2
11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + 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
-
-
-
additional information
?
-
-
metandienone hydroxylation activities and metabolism in vivo, product analysis, overview
-
-
-
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
heme
P00191
-
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
Fe2+
P00191
heme enzyme
Fe2+
-
heme enzyme
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
HgCl2
-
complete inhibition at 0.1 mM
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
Ketoconazole
-
inhibits hydroxylation activity with progesterone and 17-fluoroprogesterone to the same amount
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
-
additional information
-
no inhibition of progesterone 21-hydroxylation by the wild-type enzyme by pregnenolone, 17alpha-hydroxypregnenolone, and 5alpha-pregnane-3alpha-ol-20-one (allopregnanolone)
-
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
0.00061
-
17alpha-hydroxyprogesterone
-
pH 7.4, 37C, mutant V359G
0.00082
-
17alpha-hydroxyprogesterone
-
pH 7.4, 37C, mutant V359A
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
0.0016
-
17alpha-hydroxyprogesterone
-
pH 7.4, 37C, mutant V470A/I471A; pH 7.4, 37C, wild-type enzyme
0.00181
-
17alpha-hydroxyprogesterone
-
mutant enzyme A265V, in 50 mM potassium phosphate buffer (pH 7.4), at 37C
0.0019
-
17alpha-hydroxyprogesterone
P00191
pH 7.4, 37C, wild-type CYP21
0.002
-
17alpha-hydroxyprogesterone
-
mutant enzyme P453S
0.0024
-
17alpha-hydroxyprogesterone
P00191
pH 7.4, 37C, CYP21 mutant T241R/L442A
0.0025
-
17alpha-hydroxyprogesterone
-
pH 7.4, 37C, mutant V470A
0.0026
-
17alpha-hydroxyprogesterone
-
mutant enzyme K121Q
0.00048
-
progesterone
-
pH 7.4, 37C, mutant V359A
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
0.001
-
progesterone
-
pH 7.4, 37C, mutant V470A
0.0011
-
progesterone
-
pH 7.4, 37C, wild-type enzyme
0.0015
-
progesterone
-
mutant enzyme P453S; wild type enzyme
0.0015
-
progesterone
-
pH 7.4, 37C, mutant V470A/I471A
0.0024
-
progesterone
-
mutant enzyme K121Q
0.0032
-
progesterone
-
pH 7.4, 37C, mutant V359G
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
-
additional information
-
additional information
-
calculation of intramolecular and intermolecular kinetic isotope effects for wild-type and mutant enzymes with substrates 21-[2H]-progesterone, 21,21,21-[2H3]-progesterone, and 21,21,21-[2H3]-17-hydroxyprogesterone, overview
-
additional information
-
additional information
P00191
Pre-steady-state and steady-state binding kinetics with 17alpha-hydroxyprogesterone, stopped-flow spectroscopic measurements, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.322
-
17a-hydroxyprogesterone
-
-
0.22
-
17alpha-hydroxyprogesterone
P00191
pH 7.4, 37C, CYP21 wild-type and mutant T241R/L442A
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
-
-
7.2
-
-
assay at
7.4
-
-
-
7.4
-
-
assay at
7.4
-
P00191
assay at
7.4
-
-
assay at
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
37
-
-
assay at
37
-
P00191
assay at
37
-
-
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
additional information
P00191
structure-function analysis, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
3.6% carbohydrate
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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, 20C, few days, X-ray diffraction structure determination and analysis at 3.0 A resolution
P00191
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
-
recombinant His-tagged wild-type enzyme and mutants from Escherichia coli by nickel affinity and anion exchange chromatography, followed by gel filtration
P00191
HisTrap HP column chromatography (using Ni2+ ions immobilized onto a metal-chelating column)
-
recombinant His-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography
-
recombinant wild-type and mutant enzymes from yeast microsomes
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli
-
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
P00191
expressed in COS-1 cells
-
expressed in Sf9 insect cell microsomes
-
expressed in Spodoptera frugiperda insect cell microsomes using a baculovirus expression system
-
expression of His-tagged enzyme in Escherichia coli strain JM109, coexpression with molecular chaperones GroES and GroEL
-
expression of wild-type and mutant enzymes in yeast microsomes
-
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; gene CYP21A2, genotyping, expression of wild-type and mutant enzymes in HEK-293T and hepatoblastoma C3A cells, the H365Y enzyme is produced in more variable amounts than wild-type CYP21A2
-
mutant enzymes are expressed in COS-7 cells
-
mutant enzymes are expressed in Saccharomyces cerevisiae strain W303B
-
recombinant expression in yeast microsomes
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
T241R
P00191
site-directed mutagenesis, the mutant shows improved solubility properties compared to the wild-type enzyme
T241R/L442A
P00191
site-directed mutagenesis, the mutant shows greatly improved solubility properties compared to the wild-type enzyme
A15T
-
natural mutation found in patients with classical congenital adrenal hyperplasia, no significant difference in activity compared to wild-type
A265C
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
A265V
-
the mutant enzyme activity is similar to wild type
A265V
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
A391T
P08686
naturally occuring mutation, the mutation disrupts the hydrophobicity of the region
A434V
P08686
naturally occuring mutation, the mutation causes steric clashes with the heme rendering the enzyme almost inactive
C169R
P08686
naturally occuring mutation, the mutation alters the region's hydrophobicity, conserved residue C169 makes hydrophobic interactions with the loop between E-F helices and F-helix
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
D322G
P08686
naturally occuring mutation, the mutation prevents salt bridge formation resulting in a localized, as opposed to global, destabilization of tertiary structure
D407N
P08686
naturally occuring mutation, the mutation prevents salt bridge formation resulting in a localized, as opposed to global, destabilization of tertiary structure
E320K
P08686
naturally occuring mutation, the mutation of E320, which is a highly conserved residue on a negatively charged Glu-Glu-Leu-Asp (EELD) motif, alters the charge on the motif
E351K
-
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
E431K
P08686
naturally occuring mutation, the mutation prevents salt bridge formation resulting in a localized, as opposed to global, destabilization of tertiary structure
F404S
P08686
naturally occuring mutation, the mutation prevents stable packing interactions resulting in salt-wasting congenital adrenal hyperplasia
G178A
P08686
naturally occuring mutation, the mutation causes reduced structural flexibility of the sharp fold between the E- and F-helices
G291C
P08686
naturally occuring mutation, the mutation abolishes substrate binding causing salt-wasting congenital adrenal hyperplasia
G291R
P08686
naturally occuring mutation, the mutation abolishes substrate binding causing salt-wasting congenital adrenal hyperplasia
G291S
P08686
naturally occuring mutation, the mutation abolishes substrate binding causing salt-wasting congenital adrenal hyperplasia
G292D
P08686
naturally occuring mutation, the mutation abolishes substrate binding causing salt-wasting congenital adrenal hyperplasia
G424S
P08686
naturally occuring mutation, the mutation imparts rigidity to the loop between K'- and L-helix
G56R
P08686
naturally occuring mutation, P57 and G56 form the hinge between the membrane interacting N-terminus and rest of the protein. The substitution of G56 with a polar and rigid Arg residue disrupts the hinge affecting the interactions of CYP21A2 with the membrane
G90V
P08686
naturally occuring mutation, mutation of G90 to valine affects the ability of R91 to hydrogen bond with heme, causing salt-wasting congenital adrenal hyperplasia
H119R
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
H365W
-
the naturally occuring CYP21A2 mutant exhibits minimal 21-hydroxylase activity to convert 17-hydroxyprogesterone to 11-deoxycortisol or progesterone to 11-deoxycorticosterone compared to the wild-type
H365Y
-
the naturally occuring CYP21A2 mutant exhibits minimal 21-hydroxylase activity to convert 17-hydroxyprogesterone to 11-deoxycortisol or progesterone to 11-deoxycorticosterone compared to the wild-type
H365Y
P08686
naturally occuring mutation, the mutations causes the salt-wasting disease
H365Y/R356W
-
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; heterozygote H365Y/R356W individuum for two CYP21A2 gene mutations each inherited from a different parent
H62L
P08686
naturally occuring mutation, the mutation may disrupt hydrogen bonding to reduce, but not eliminate, enzyme activity
I171N
-
mutation identified in Italian patient with congenital adrenal hyperplasia, less than 1% of wild-type enzyme activity
I172N
-
naturally occuring mutant, 0-2% of wild-type activity, dominant negative effect over wild-type with 11% decrease in activity
I172N
-
the mutation is associated with congenital adrenal hyperplasia
I172N
P08686
naturally occuring mutation, the mutation causes a loss of the hydrophobic pocket, I172 forms a hydrophobic pocket with M186 and M187 residues of F-helix
I194N
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
I230T
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
I236N/V237E/M239K
-
naturally occuring mutant, no enzymic activity, dominant negative effect over wild-type with 35% decrease in activity
I471A
-
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
I471G
-
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
I77T
P08686
naturally occuring mutation, the mutation disrupts the hydrophobic environment
K121Q
P08686
naturally occuring mutation, the mutation impairs the interaction with the P450 oxidoreductase
K122Q
-
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
L107R
P08686
naturally occuring mutation, the mutation abolishes heme binding and causes salt-wasting congenital adrenal hyperplasia
L107X
-
site-directed mutagenesis, inactive mutant
L109X
-
site-directed mutagenesis, inactive mutant
L142P
P08686
naturally occuring mutation, the mutation of the D-helix causes helical disruption and destabilization of secondary structures
L166P
P08686
naturally occuring mutation, the mutation of the E-helix causes helical disruption and destabilization of secondary structures
L167P
P08686
naturally occuring mutation, the mutation of the E-helix causes helical disruption and destabilization of secondary structures
L236N/V237E/M239K
-
the mutation is associated with congenital adrenal hyperplasia
L261P
P08686
naturally occuring mutation, the mutation of the H-helix causes helical disruption and destabilization of secondary structures
L300F
P08686
naturally occuring mutation, the mutation causes localized destabilization of secondary structure
L307M
P08686
naturally occuring mutation, the mutation disrupts the optimal packing of side chains but does not alter the hydrophobic environment
L307V
P08686
naturally occuring mutation, the mutation disrupts the optimal packing of side chains but does not alter the hydrophobic environment
L308F
P08686
naturally occuring mutation, the mutation causes localized destabilization of secondary structure
L353R
P08686
naturally occuring mutation, the mutation abolishes heme binding and causes salt-wasting congenital adrenal hyperplasia
L363W
P08686
naturally occuring mutation, the mutation causes steric clashes with the heme rendering the enzyme almost inactive
L446P
-
mutation identified in Italian patient with congenital adrenal hyperplasia, less than 1% of wild-type enzyme activity
N387K
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
P30E
P08686
naturally occuring mutation causing disruption of the interaction between the carbon of P30 in the N-terminal loop and the side chain of Y376 within the beta5-beta6 hairpin loop resuting in the salt-wasting disease
P30L
-
missense mutation associated with congenital adrenal hyperplasia
P30L
-
the mutation is associated with congenital adrenal hyperplasia
P432L
P08686
naturally occuring mutation, the mutation makes the structure more flexible and prevents cysteine from being presented to heme
P453S
-
the mutant shows a reduced activity of 36% of wild type for the conversion of 17alpha-hydroxyprogesterone and 44% for the conversion of progesterone
P453S
P08686
naturally occuring mutation, the mutation disrupts the hydrophobicity of the region
P459H
P08686
naturally occuring mutation, the mutation disrupts the hydrophobicity of the region
P463L
P08686
naturally occuring mutation, the mutation interferes with the conformation of the beta8-beta9 loop with the subsequent closure of substrate entrance channel
P482S
-
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
-
the mutation is associated with congenital adrenal hyperplasia
Q481P
P08686
naturally occuring mutation, the mutation destabilizes the structure rendering the protein inactive
R124H
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
R132C
P08686
naturally occuring mutation, the mutation impairs the interaction with the P450 oxidoreductase
R149C
P08686
naturally occuring mutation, the mutation prevents salt bridge formation resulting in a localized, as opposed to global, destabilization of tertiary structure
R233G
P08686
naturally occuring mutation, the mutation may prevent R233 from binding to the 3-keto oxygen of the proximal 17OHP in the proper orientation, it does not influence protein activity significantly, resulting in minimal phenotype
R233K
P08686
naturally occuring mutation, the mutation may prevent R233 from binding to the 3-keto oxygen of the proximal 17OHP in the proper orientation, it does not influence protein activity significantly, resulting in minimal phenotype
R339H
P08686
naturally occuring mutation, the mutation impairs the interaction with the P450 oxidoreductase
R341P
-
mutation identified in Italian patient with congenital adrenal hyperplasia, less than 1% of wild-type enzyme activity
R341P
P08686
naturally occuring mutation, the mutation impairs the interaction with the P450 oxidoreductase
R341W
P08686
naturally occuring mutation, the mutation impairs the interaction with the P450 oxidoreductase
R356P
P08686
naturally occuring mutation, the mutation disrupts the interaction of R356 with Q389 rendering the enzyme inactive and causing salt-wasting congenital adrenal hyperplasia
R356W
-
naturally occuring mutant, no enzymic activity, no dominant negative effect on wild-type
R356W
-
the mutation is associated with congenital adrenal hyperplasia
R369Q
P08686
naturally occuring mutation, the mutation impairs the interaction with the P450 oxidoreductase
R408C/L
P08686
naturally occuring mutation, the mutation destabilizes structural elements because of the extensive loss of hydrogen bonds
R408H
P08686
naturally occuring mutation, the mutation prevents normal hydrogen bonding with E351 and R354
R426C
P08686
naturally occuring mutation, the mutation disrupts the interaction of residues R91 and R426 rendering the protein nonfunctional and causing salt-wasting congenital adrenal hyperplasia
R426H
-
mutation identified in Italian patient with congenital adrenal hyperplasia, less than 1% of wild-type enzyme activity
R435C
P08686
naturally occuring mutation, the mutation prevents salt bridge formation resulting in a localized, as opposed to global, destabilization of tertiary structure
R479L
P08686
naturally occuring mutation, the mutation prevents salt bridge formation resulting in a localized, as opposed to global, destabilization of tertiary structure
R483P
P08686
naturally occuring mutation, the mutation prevents salt bridge formation resulting in a localized, as opposed to global, destabilization of tertiary structure
R483Q
-
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
R483Q
P08686
naturally occuring mutation, the mutation prevents salt bridge formation resulting in a localized, as opposed to global, destabilization of tertiary structure
S301Y
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
T168N
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
T295X
P08686
naturally occuring mutation, the mutation abolishes substrate binding and causes salt-wasting congenital adrenal hyperplasia
T450P
P08686
naturally occuring mutation, the mutation reduces flexibility of beta8-sheet, which helps stabilize the very long C-terminal loop
V139E
P08686
naturally occuring mutation, mutation to glutamate disrupts the interaction with residues V440 and L436 on the L-helix causing instability of the enzyme, charge repulsions between the side chain of mutated V139E and E437 of the E-helix render the protein unstable and inactive causing salt-wasting congenital adrenal hyperplasia
V249A
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
V281G
P08686
naturally occuring mutation, the mutation causes a loss of the hydrophobic pocket
V281L
-
naturally occuring mutant, 50% of wild-type activity, dominant negative effect over wild-type with 30% decrease in activity
V281L
-
the mutation is associated with congenital adrenal hyperplasia
V304M
P08686
naturally occuring mutation, the mutation causes side-chain steric clashes with the neighboring residues
V359A
-
site-directed mutagenesis, the mutant shows 60% reduced activity compared to the wild-type enzyme
V359A
-
site-directed mutagenesis, the mutant shows altered product formation compared to the wild-type enzyme
V359G
-
site-directed mutagenesis, the mutant shows 10% reduced activity compared to the wild-type enzyme
V470A
-
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
V470A/I471A
-
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
V470G
-
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
W302R
P08686
naturally occuring mutation, the mutation prevents stable packing interactions resulting in salt-wasting congenital adrenal hyperplasia
W302S
-
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
W302S
P08686
naturally occuring mutation, the mutation prevents stable packing interactions resulting in salt-wasting congenital adrenal hyperplasia
Y47C
P08686
naturally occuring mutation, the mutation disables hydrogen bonding with H38, the interaction is compensated by a weak His-Cys interaction
Y47L
P08686
naturally occuring mutation, the mutation disrupts hydrogen bonds and causes salt-wasting congenital adrenal hyperplasia
Y59N
P08686
naturally occuring mutation, the mutation disrupts the hydrophobicity of the region resulting in loss of function
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
P08686
naturally occuring mutation, the mutation of the L-helix causes helical disruption and destabilization of secondary structures
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