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Information on EC 1.14.15.6 - cholesterol monooxygenase (side-chain-cleaving) and Organism(s) Bos taurus and UniProt Accession P00189

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IUBMB Comments
A heme-thiolate protein (cytochrome P-450). The reaction proceeds in three stages, with two hydroxylations at C-22 and C-20 preceding scission of the side-chain between carbons 20 and 22. The initial source of the electrons is NADPH, which transfers the electrons to the adrenodoxin via EC 1.18.1.6, adrenodoxin-NADP+ reductase.
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Bos taurus
UNIPROT: P00189
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The taxonomic range for the selected organisms is: Bos taurus
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
hide(Overall reactions are displayed. Show all >>)
cholesterol
+
6
reduced adrenodoxin
+
3
O2
+
6
H+
=
pregnenolone
+
4-methylpentanal
+
6
oxidized adrenodoxin
+
4
H2O
+
6
reduced adrenodoxin
+
3
+
6
=
+
+
6
oxidized adrenodoxin
+
4
Synonyms
p450scc, cyp11a1, cytochrome p-450scc, cytochrome p450scc, cyp11a, cholesterol side-chain cleavage enzyme, p450(scc), cholesterol side chain cleavage enzyme, cholesterol side-chain cleavage cytochrome p450, cytochrome p450 cholesterol side-chain cleavage, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cholesterol side chain cleavage enzyme
-
cytochrome P450 11A1
-
P450 11A1
-
C27-side chain cleavage enzyme
-
-
-
-
cholesterol 20-22-desmolase
-
-
-
-
cholesterol C20-22 desmolase
-
-
-
-
cholesterol C20-C22 lyase
-
-
-
-
cholesterol desmolase
-
-
-
-
cholesterol hydroxylase
-
-
cholesterol side-chain cleavage enzyme
cholesterol side-chain-cleaving enzyme
-
-
-
-
CYPXIA1
-
-
-
-
cytochrome P-450scc
-
-
-
-
cytochrome P450 cholesterol side chain cleavage
-
-
cytochrome P450scc
-
-
desmolase, steroid 20-22
-
-
-
-
endoenzymes, cholesterol side-chain-cleaving
-
-
-
-
enzymes, cholesterol side-chain-cleaving
-
-
-
-
P450 cholesterol side-chain cleavage enzyme
-
-
P450(scc)
-
-
-
-
steroid 20-22 desmolase
-
-
-
-
steroid 20-22-lyase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
cholesterol,reduced-adrenal-ferredoxin:oxygen oxidoreductase (side-chain-cleaving)
A heme-thiolate protein (cytochrome P-450). The reaction proceeds in three stages, with two hydroxylations at C-22 and C-20 preceding scission of the side-chain between carbons 20 and 22. The initial source of the electrons is NADPH, which transfers the electrons to the adrenodoxin via EC 1.18.1.6, adrenodoxin-NADP+ reductase.
CAS REGISTRY NUMBER
COMMENTARY hide
37292-81-2
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(20R,22R)-20,22-dihydroxy-cholesterol + 2 reduced adrenodoxin + O2
pregnenolone + 4-methylpentanal + 2 oxidized adrenodoxin + 2 H2O
show the reaction diagram
-
-
-
?
(22R)-22-hydroxycholesterol + 2 reduced adrenodoxin + O2
(20R,22R)-20,22-dihydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
cholesterol + 2 reduced adrenodoxin + O2
(22R)-22-hydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
?
cholesterol + 6 reduced adrenodoxin + 3 O2
pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O
show the reaction diagram
overall reaction
-
-
?
(20S)-22-thiacholesterol + reduced adrenodoxin + O2
(20S,22R)-22-thiacholesterol S-oxide + (20S,22S)-22-thiacholesterol S-oxide
show the reaction diagram
-
-
(22R)-sulfoxide preferentially formed by a factor of 4.2 to 1 over (22S)-sulfoxide
?
17,20-dihydroxyvitamin D2 + reduced adrenodoxin + O2
17,20,24-trihydroxyvitamin D2 + oxidized adrenodoxin + O2
show the reaction diagram
-
is slowly metabolized
-
-
?
20,23-dihydroxyvitamin D3 + reduced adrenodoxin + O2
17,20,23-trihydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20,23-dihydroxyvitamin D3 + reduced adrenodoxin + O2
17alpha,20,23-trihydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20-hydroxyvitamin D2 + reduced adrenodoxin + O2
17,20,24-trihydroxyvitamin D2 + oxidized adrenodoxin + O2
show the reaction diagram
-
is a better substrate than vitamin D2 itself
-
-
?
20-hydroxyvitamin D3 + reduced adrenodoxin + O2
20,23-dihydroxyvitamin D3 + 17alpha,20,23-trihydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20-hydroxyvitamin D3 + reduced adrenodoxin + O2
20,23-dihydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20alpha, 22(R)-dihydroxycholesterol + reduced adrenodoxin + O2
? + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
20alpha-hydroxycholesterol + reduced adrenodoxin + O2
pregnenolone + 4-methylpentanal + oxidized adrenodoxin + H2O
show the reaction diagram
22(R)-hydroxycholesterol + reduced adrenodoxin + O2
? + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
22(R)-hydroxycholesterol + reduced adrenodoxin + O2
pregnenolone + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
cholesterol + 6 reduced adrenodoxin + 3 O2
pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O
show the reaction diagram
cholesterol + reduced adrenal ferredoxin + O2
pregnenolone + 4-methylpentanal + oxidized adrenal ferredoxin
show the reaction diagram
-
-
-
-
?
cholesterol + reduced adrenodoxin + O2
? + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
?
cholesterol + reduced adrenodoxin + O2
pregnenolone + 4-methylpentanal + oxidized adrenodoxin + H2O
show the reaction diagram
cholesterol + reduced adrenodoxin + O2 + H+
pregnenolone + 4-methylpentanal + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
-
r
cholesterol + reduced ferredoxin + O2
pregnenolone + 4-methylpentanal + oxidized ferredoxin
show the reaction diagram
cholesterol sulfate + reduced adrenodoxin + O2
pregnenolone sulfate + 17-hydroxy-pregnenolone + dehydroisoandrosterone sulfate + oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
?
vitamin D2 + reduced adrenodoxin + O2
17,20,24-trihydroxyvitamin D2 + oxidized adrenodoxin + H2O
show the reaction diagram
-
P450scc catalyzes three sequential hydroxylations of D2 producing 20-hydroxyvitamin D2, 17,20-dihydroxyvitamin D2, and 17,20,24-trihydroxyvitamin D2, which dissociate from the active site of P450scc and accumulate in the reaction mixture
-
-
?
vitamin D3 + reduced adrenodoxin + O2
20-hydroxyvitamin D3 + oxidized adrenodoxin + H2O
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(20R,22R)-20,22-dihydroxy-cholesterol + 2 reduced adrenodoxin + O2
pregnenolone + 4-methylpentanal + 2 oxidized adrenodoxin + 2 H2O
show the reaction diagram
-
-
-
?
(22R)-22-hydroxycholesterol + 2 reduced adrenodoxin + O2
(20R,22R)-20,22-dihydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
?
cholesterol + 2 reduced adrenodoxin + O2
(22R)-22-hydroxycholesterol + 2 oxidized adrenodoxin + H2O
show the reaction diagram
-
-
-
?
cholesterol + 6 reduced adrenodoxin + 3 O2
pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O
show the reaction diagram
overall reaction
-
-
?
cholesterol + 6 reduced adrenodoxin + 3 O2
pregnenolone + 4-methylpentanal + 6 oxidized adrenodoxin + 4 H2O
show the reaction diagram
cholesterol + reduced adrenal ferredoxin + O2
pregnenolone + 4-methylpentanal + oxidized adrenal ferredoxin
show the reaction diagram
-
-
-
-
?
cholesterol + reduced ferredoxin + O2
pregnenolone + 4-methylpentanal + oxidized ferredoxin
show the reaction diagram
-
metabolic regulation of enzyme expression, overview
-
-
?
additional information
?
-
-
the enzyme is important in steroidogenesis, regulatory mechanism, overview
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adrenodoxin
-
-
cytochrome P450
-
-
heme
site of catalysis
adrenodoxin
-
cytochrome P450
-
-
-
Ferredoxin
-
-
-
NADPH
reduced adrenal ferredoxin
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
-
cytochrome P450 enzyme
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(20R,S)-20-amino-5-pregnen-3beta-ol
-
20-amine derivative, amine is attached closer to the D-ring than in the 22-amine, very weak inhibitor, 0.1 mM causes less than 20% inhibition
(20S)-22-thiacholesterol
-
competitive inhibitor, is converted enzymatically to a more potent inhibitor, the (22S) and (22R) sulfoxides, inhibition by approximately 75% at 0.001 mM, no inactivation in absence of NADPH and O2
(20S,22R)-22-thiacholesterol S-oxide
-
competitive versus cholesterol, binds 10times more tightly than (22S) diastereomer, 25% and 44% inhibition at 0.00005 mM and 0.0001 mM, respectively, complete inhibition at 0.001 mM, no substrate for P-450
(20S,22S)-22-thiacholesterol S-oxide
-
competitive versus cholesterol, no substrate for P-450
(22R)-22-Aminocholesterol
-
completely inhibited by 0.001 mM, affinity toward the P-450scc almost 3fold greater than that for the (22S) form, competitive versus cholesterol, no substrate for P-450
(22S)-22-Aminocholesterol
-
not inhibited below 0.001 mM, competitive versus cholesterol, no substrate for P-450
17,20-dihydroxyvitamin D2
-
competitive inhibitor
17beta-amino-5-androsten-3beta-ol
-
17-amine derivative, amine is attached closer to the D-ring than in the 22-amine, very weak inhibitor, 0.1 mM causes less than 20% inhibition
22-Amino-23,24-bisnor-5-cholen-3beta-ol
22-amino-23,24-bisnor-5alpha-cholen-3beta-ol
-
50% inhibition at 0.003 mM
23,24-bisnor-5-cholene-3beta,22-diol
-
competitive inhibitor, 40% inhibition at 0.01 mM, 50% at 0.015 mM, resembles the intermediate 22-hydroxycholesterol but acts as an inhibitor rather than serving as a substrate
23-Amino-24-nor-5-cholen-3beta-ol
-
23-amine derivative, same steroid ring structure as cholesterol, competitive inhibitor with respect to cholesterol, 50% inhibition at 0.0001 mM, reversible cooperative binding
24-Amino-5-cholen-3beta-ol
-
24-amine derivative, amine attached in greater distance from steroid ring, same steroid ring structure as cholesterol, causes a progressive decrease in inhibitory potency, 50% inhibition at 0.0023 mM, reversible noncooperative binding
25-Amino-26,27-bisnor-5-cholesten-3beta-ol
-
25-amine derivative, amine attached in greater distance from steroid ring, causes a progressive decrease in inhibitory potency, 50% inhibition at more than 0.1 mM
adrenodoxin
-
oxidized form, high affinity to P-450scc, inhibits side chain cleavage by competition with reduced form
-
cholesterol
-
inhibition above 0.003 mM, mitochondrial
Cholesterol sulfate
-
inhibition above 0.005 mM, mitochondrial
glycerol
-
substrate cholesterol, 50% inhibition at 25% glycerol, substrate 22(R)-hydroxycholesterol, 50% inhibition at 44% glycerol, substrate 20alpha-hydroxycholesterol, 50% inhibition at 48% glycerol, substrate 20alpha, 22(R)-dihydroxycholesterol, 50% inhibition at 51% glycerol
methoxychlor
-
pesticide of DDT, suicide inhibitor, competitive to cholesterol, substantial irreversible loss of activity, 5% inhibition within 5 min at 0.2 mM, decrease is suppressed by the presence of cholesterol
phosphatidyl choline
-
-
phosphatidyl ethanolamine
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,2-di-(2'-hexyl-decanoyl)-sn-glycero-3-phosphocholine
-
alpha-branched phosphatidylcholine, inclusion in vesicle-reconstituted system, partially in connection with the nonactivator lipids dimyristoyl-/dioleoyl-phosphatidylcholine, efficiency close to cardiolipin
1,2-di-(2'-octyl-dodecanoyl)-sn-glycero-phosphocholine
-
alpha-branched phosphatidylcholine, inclusion in vesicle-reconstituted system, partially in connection with the nonactivator lipids dimyristoyl-/dioleoyl-phosphatidylcholine, efficiency close to cardiolipin
Brij
-
56, 76 and 96
-
cardiolipin
Emulgen
-
911 and 913
-
fatty acid
-
C18, natural detergents in DMPC vesicles, stimulation similar to octyl glucoside
glycerol
-
5-20% concentration, 20-50% increase of enzyme activity
Lipid
-
from adrenal mitochondria accelerates activity
octyl glucoside
-
high concentrations of this detergent cause 50% stimulation of cholesterol side-chain-cleavage in large unilamellar vesicles at low cholesterol concentration, 0.01 mM increase the proportion of P-450 bound by cholesterol
Phospholipid
Triton X-100
-
effect onto P-450 itself suggested
Tween 20
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0181
17,20-dihydroxyvitamin D2
-
in 0.45% cyclodextrin
0.012
20-hydroxyvitamin D2
-
in 0.45% cyclodextrin
0.067
20-hydroxyvitamin D3
-
in the presence of 0.45% cyclodextrin
0.012
20alpha-hydroxycholesterol
-
-
0.0005 - 1.18
cholesterol
0.0003 - 0.0232
Cholesterol sulfate
0.0004 - 0.0012
reduced adrenodoxin
-
0.0175
vitamin D2
-
in 0.45% cyclodextrin
0.0296 - 3.67
vitamin D3
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.123
20-hydroxyvitamin D3
-
in the presence of 0.45% cyclodextrin
0.05 - 0.5
cholesterol
0.328 - 1.6
vitamin D3
additional information
additional information
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.83
20-hydroxyvitamin D3
-
in the presence of 0.45% cyclodextrin
0.212 - 11.17
cholesterol
0.435 - 11.1
vitamin D3
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8
-
for cholesterol and 20alpha-hydroxycholesterol, 75 mM potassium phosphate buffer
7.4
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7.5
-
less than 50% of maximal activity below and above range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 42
-
tested in different dimyristoyl-phosphatidylcholine vesicles, breaks in activity at 27-30°C
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
granulosa cell
Manually annotated by BRENDA team
additional information
-
immunohistochemic localization analysis, overview
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
the activity of P450scc is dependent upon electron delivery from NADPH-dependent adrenodoxin reductase (AdR), via adrenodoxin (Adx). The lipid membrane assists in the assembly of electron transport proteins and the activity of P450scc by providing a surface for the localized concentration of proteins. Using sequential deposition with P450scc first (to saturation), there is effectively no further binding of AdR or Adx. When premixing the three proteins, P450scc, Adx and AdR, all bind the membrane in a two-stage process
metabolism
-
vitamin D2 metabolism occurs with lower efficiency than that observed for both cholesterol and D3 metabolism by P450scc
physiological function
-
Escherichia coli membrane interacting peptides in CYP11A1 are from the putative F-G loop (residues 218-225) and the C-terminal portion of the G helix (residues 238-250)
additional information
the active site cavity in CYP11A1 represents a long curved tube that extends from the protein surface to the heme group, the site of catalysis. Shuttling of the sterol intermediates between the active site entrance and the heme group during the three-step reaction. Structural basis of the strict substrate specificity and high catalytic efficiency of the enzyme, overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
CP11A_BOVIN
520
0
60333
Swiss-Prot
Mitochondrion (Reliability: 4)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
100000
-
native-PAGE, incomplete enzyme or molecular environmental conditions not optimal
120000
-
x * 120000, fusion protein of enzyme plus adrenodoxin reductase plus adrenodoxin
200000
220000
-
corpus luteum
225000
-
sedimentation equilibrium
400000
-
native-PAGE
415000
-
sedimentation equilibrium
46000
470000
-
sedimentation equilibrium performed in 100 mM potassium phosphate buffer, pH 7.6
48000
-
2,4,6,8 * 48000, SDS-PAGE, corpus luteum. x * 46000, x * 60000, x * 53000, x * 52000-53000 adrenal mitochondria
52000
53000
60000
800000
-
exclusion chromatography
850000
97000
-
sedimentation equilibrium
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexadecamer
-
16 * 53000, sedimentation equilibrium performed in 6 M guanidine after heating, 16 * 52000, SDS-PAGE. Can also exist in forms of 4 and 8 subunits after treatment with 100 mM potassium phosphate buffer, 7.6, with molecular weights of 200000 and 470000, respectively
tetramer
-
4 * 46000, sedimentation equilibrium analysis after guanidine treatment and SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CYP11A1 in complex with (22R)-22-hydroxycholesterol, sitting drop vapour diffusion methdo, mixing of 0.001 ml of 23 mg/ml protein in 50 mM potassium phosphate, pH 7.2, 20% glycerol, 0.1 M NaCl, 0.1% octyl pentaethylene glycol ether, 1 mM EDTA, and 0.05 mM (22R)-22-hydroxycholesterol, with 0.001 ml of precipitant solution containing 14% PEG 1000, 20% glycerol, 12% JEFFAMINE ED-2001, 0.1 M MES, pH 7.0, and 10% isopropyl alcohol, 18°C, overnight, X-ray diffraction structure determination and analysis at 2.8 A resolution, molecular replacement using rat mitochondrial CYP24A1
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K103Q
K104Q
K109Q
K110Q
K145Q
K148Q
K267Q
-
participation in electrostatic interaction of P-450scc with adrenodoxin
K338Q
-
removed from heme group but still very important for interaction with adrenodoxin, K helix
K342Q
-
removed from heme group but still very important for interaction with adrenodoxin, K helix
K394Q
K403Q
K405Q
R425Q
-
most harmful substitution, L helix, heme-binding region
R425Q/R426C
-
double mutant, most harmful substitution
R425Q/R426Q
-
double mutant, most harmful substitution
R426Q
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
increase of complex formation of adrenodoxin and cytochrome P-450scc in absence of cholesterol with decrease of temperature to 6°C, elevated temperature decreases the affinity of P-450scc for cholesterol
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
catalase, protects P-450scc against activity loss
-
glycerol, 50%, stabilizes
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Glycerol
-
induces sigmoidal low-spin response of otherwise high-spin spectrum, weakens adrenodoxin binding, inhibitory effect being substrate dependent
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 10 mM sodium phosphate buffer, pH 7.4, 0.1 mM EDTA, 20% glycerol, stable for months
-
-20°C, 50 mM potassium phosphate buffer, pH 7.0, 50% glycerol, 3 months
-
-20°C, 50% glycerol, for months
-
-20°C, mitochondrial precipitate, 10 mM sodium phosphate buffer, pH 7.4, 0.1 mM EDTA
-
-20°C, purified, 10 mM sodium phosphate buffer, pH 7.4, 0.1 mM EDTA, 20% glycerol, for months without loss of P-450 activity
-
-70°C, mitochondrial pellet, 30-40 mg/ml, 100 mM potassium phosphate, pH 7.3, 0.2 mM EDTA
-
-80°C, 50 mM potassium phosphate buffer, pH 7.0, 0.1 mM EDTA, 0.1 mM dithiothreitol, 0.01% cholate, a few days without substantial loss of activity
-
0°C on ice, purified, 20 mM potassium phosphate buffer, pH 7.4, 20% glycerol, v/v, 0.1 mM EDTA, 0.1 mM dithioerythritol, 0.01% Emulgen 911, 3 weeks, without substantial loss of activity
-
4°C, purified, 50 mM potassium phosphate, pH 7.3, 0.1 mM dithiothreitol, 10% glycerol, stable a few weeks
-
5°C, 50 mM potassium phosphate buffer, pH 7.0, without glycerol, 21-43% decrease of activity within 30 days, completely lost in 4 months
-
purified, 25 mM potassium phosphate, pH 7.3, 0.05 mM EDTA, 0.05 mM dithiothreitol, 0.025 mM deoxycorticosterone, 0.25% Tween 20, 0.25% sodium cholate
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant C-terminally His4-tagged CYP11A1 by anion exchange, nickel affinity, hydroxylapatite, adrenodoxin affinity chromatography, and gel filtration
affinity chromatography
-
ammonium sulfate fractioning, aniline-Sepharose, adrenodoxin-Separose
-
ammonium sulfate precipitation, affinity chromatography
-
cholate extraction, pyrophosphate treatment, affinity chromatography
-
cholic acid extraction, ammonium sulfate precipitation, DEAE-cellulose, hydroxylapatite and gel filtration
-
column chromatography, gel filtration
-
DEAE-cellulose, adrenodoxin-Sepharose
-
iso-octane and ammonium sulfate fractionating, gel filtration
-
precipitation with polyethylene glycol, affinity chromatography
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
several ion exchange and heptyl-Sepharose hydrophobic chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli and Saccharomyces cerevisiae
recombinant expression of C-terminally His4-tagged CYP11A1
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
heterologous expression in Escherichia coli
-
mutants and wild-type expressed in Escherichia coli
-
Nicotiana tabacum plants expressing CYP11A1 cDNA
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
contents of soluble protein and carbohydrates in leaves and seeds of transgenic Nicotiana tabacum plants are essentially higher than the contents of these components in leaves and seeds of control plants
-
CYP11A1 is elevated in dominant follicles. CYP11A1 mRNA levels are highly correlated with the focimatrix genes COL4A1, NID1 and -2 and HSPG2. Focimatrix may potentially regulate CYP11A1 expression, and the regulation of both may be important in follicular dominance
-
mRNA levels for CYP11A1 are significantly lower in subordinate follicles in comparison to dominant follicles and this effect is maintained following adjustment for follicle size
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
examination of protein-membrane interactions between P450scc and its redox partners on 1,2-dimyristoyl-sn-glycero-3-phosphocholine membranes containing cholesterol (20%), using a quartz crystal microbalance with dissipation monitoring
synthesis
construction of a fusion protein consisting of cytochrome P450scc (CYP11A1), adrenodoxin and adrenodoxin reductase including 2A peptide from Picornaviridae which is capable of self-cleavage. Introduction to Escherichia coli leads to a high level of expression but no cleavage. In yeast Saccharomyces cerevisiae, the discrete proteins P450scc-2A, adrenodoxin-2A and adrenodoxin reductase are expressed, with a significant proportion present in a fusion adrenodoxin-2A-adrenodoxin reductase. The enzyme system is catalytically active
drug development
-
the vitamin D3 derivatives produced by the action of P450scc are good candidates for use in the therapy of hyperproliferative disorders
medicine
-
treatment of certain hormone-related pathologies
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Hanukoglu, I.; Jefcoate, C.R.
Mitochondrial cytochrome P-450sec. Mechanism of electron transport by adrenodoxin
J. Biol. Chem.
255
3057-3061
1980
Bos taurus
Manually annotated by BRENDA team
Hanukoglu, I.; Spitsberg, V.; Bumpus, J.A.; Dus, K.M.; Jefcoate, C.R.
Adrenal mitochondrial cytochrome P-450scc. Cholesterol and adrenodoxin interactions at equilibrium and during turnover
J. Biol. Chem.
256
4321-4328
1981
Bos taurus
Manually annotated by BRENDA team
Hanukoglu, I.; Privalle, C.T.; Jefcoate, C.R.
Mechanisms of ionic activation of adrenal mitochondrial cytochromes P-450scc and P-45011beta
J. Biol. Chem.
256
4329-4335
1981
Bos taurus
Manually annotated by BRENDA team
Wang, H.P.; Kimura, T.
Purification and characterization of adrenal cortex mitochondrial cytochrome P-450 specific for cholesterol side chain cleavage activity
J. Biol. Chem.
251
6068-6074
1976
Bos taurus
Manually annotated by BRENDA team
Nakajin, S.; Ishii, Y.; Shinoda, M.
Binding of Triton X-100 to purified cytochrome P-450scc and enhancement of the cholesterol side chain cleavage activity
Biochem. Biophys. Res. Commun.
87
524-531
1979
Bos taurus
Manually annotated by BRENDA team
Hume, R.; Kelly, R.W.; Taylor, P.L.; Boyd, G.S.
The catalytic cycle of cytochrome P-450scc and intermediates in the conversion of cholesterol to pregnenolone
Eur. J. Biochem.
140
583-591
1984
Bos taurus
Manually annotated by BRENDA team
Shikita, M.; Hall, P.F.
Cytochrome P-450 from bovine adrenocortical mitochondria: an enzyme for the side chain cleavage of cholesterol. I. Purification and properties
J. Biol. Chem.
248
5598-5604
1973
Bos taurus
Manually annotated by BRENDA team
Takemori, S.; Sukara, K.; Hashimoto, K.; Hashimoto, M.; Sato, H.; Gomi, T.; Katagiri, M.
Purification of cytochrome P-450 from bovine adrenocortical mitochondria by an aniline-Sepharose and the properties
Biochem. Biophys. Res. Commun.
63
588-593
1975
Bos taurus
Manually annotated by BRENDA team
Dhariwal, M.S.; Jefcoate, C.R.
Cholesterol metabolism by purified cytochrome P-450scc is highly stimulated by octyl glucoside and stearic acid exclusively in large unilamellar phospholipid vesicles
Biochemistry
28
8397-8402
1989
Bos taurus
Manually annotated by BRENDA team
Lambeth, J.D.; Kriengsiri, S.
Cytochrome P-450scc-adrenodoxin interactions. Ionic effects on binding, and regulation of cytochrome reduction by bound steroid substrates
J. Biol. Chem.
260
8810-8816
1985
Bos taurus
Manually annotated by BRENDA team
Nagahisa, A.; Foo, T.; Gut, M.; Orme-Johnson, W.H.
Competitive inhibition of cytochrome P-450scc by (22R)- and (22S)-22-aminocholesterol. Side-chain stereochemical requirements for C-22 amine coordination to the active-site heme
J. Biol. Chem.
260
846-851
1985
Bos taurus
Manually annotated by BRENDA team
Kashiwagi, K.; Dafeldecker, W.P.; Salhanick, H.A.
Purification and characterization of mitochondrial cytochrome P-450 associated with cholesterol side chain cleavage from bovine corpus luteum
J. Biol. Chem.
255
2606-2611
1980
Bos taurus
Manually annotated by BRENDA team
Sheets, J.J.; Vickery, L.E.
Active site-directed inhibitors of cytochrome P-450scc. Structural and mechanistic implications of a side chain-substituted series of amino-steroids
J. Biol. Chem.
258
11446-11452
1983
Bos taurus
Manually annotated by BRENDA team
Sheets, J.J.; Vickery, L.E.
C-22-Substituted steroid derivatives as substrate analogues and inhibitors of cytochrome P-450scc
J. Biol. Chem.
258
1720-1725
1983
Bos taurus
Manually annotated by BRENDA team
Warne, P.A.; Greenfield, N.J.; Lieberman, S.
Modulation of the kinetics of cholesterol side-chain cleavage by an activator and by an inhibitor isolated from the cytosol of the cortex of bovine adrenals
Proc. Natl. Acad. Sci. USA
80
1877-1881
1983
Bos taurus
Manually annotated by BRENDA team
Shikita, M.; Hall, P.F.
Cytochrome P-450 from bovine adrenocortical mitochondria: an enzyme for the side chain cleavage of cholesterol. II. Subunit structure
J. Biol. Chem.
248
5605-5609
1973
Bos taurus
Manually annotated by BRENDA team
Miao, E.; Joardar, S.; Zuo, C.; Cloutier, N.J.; Nagahisa, A.; Byon, C.; Wilson, S.R.; Orme-Johnson, W.H.
Cytochrome P-450scc-mediated oxidation of (20S)-22-thiacholesterol: Characterization of mechanism-based inhibition
Biochemistry
34
8415-8421
1995
Bos taurus
Manually annotated by BRENDA team
Schwarz, D.; Kisselev, P.; Wessel, R.; Jueptner, O.; Schmid, R.D.
alpha-Branched 1,2-diacyl phosphatidylcholines as effectors of activity of cytochrome P450SCC (CYP11A1). Modeling the structure of the fatty acyl chain region of cardiolipin
J. Biol. Chem.
271
12840-12846
1996
Bos taurus
Manually annotated by BRENDA team
Lepesheva, G.I.; Azeva, T.N.; Strushkevich, N.V.; Gilep, A.A.; Usanov, S.A.
Site-directed mutagenesis of cytochrome P450scc (CYP11A1). Effect of lysine residue substitution on its structural and functional properties
Biochemistry (Moscow)
65
1409-1418
2000
Bos taurus
Manually annotated by BRENDA team
Ahmed, S.
The use of the novel substrate-heme complex approach in the derivation of a representation of the active site of the enzyme cholesterol side chain cleavage
Biochem. Biophys. Res. Commun.
274
821-824
2000
Bos taurus
Manually annotated by BRENDA team
Tsujita, M.; Ichikawa, Y.
Substrate-binding region of cytochrome P-450scc (P-450 XIA1). Identification and primary structure of the cholesterol binding region in cytochrome P-450scc
Biochim. Biophys. Acta
1161
124-130
1993
Bos taurus
Manually annotated by BRENDA team
Usanov, S.A.; Graham, S.E.; Lepesheva, G.I.; Azeva, T.N.; Strushkevich, N.V.; Gilep, A.A.; Estabrook, R.W.; Peterson, J.
Probing the interaction of bovine cytochrome P450scc (CYP11A1) with adrenodoxin: evaluating site-directed mutations by molecular modeling
Biochemistry
41
8310-8320
2002
Bos taurus
Manually annotated by BRENDA team
Headlam, M.J.; Tuckey, R.C.
The effect of glycerol on cytochrome P450scc (CYP11A1) spin state, activity, and hydration
Arch. Biochem. Biophys.
407
95-102
2002
Bos taurus
Manually annotated by BRENDA team
Nazarov, P.A.; Drutsa, V.L.; Miller, W.L.; Shkumatov, V.M.; Luzikov, V.N.; Novikova, L.A.
Formation and functioning of fused cholesterol side-chain cleavage enzymes
DNA Cell Biol.
22
243-252
2003
Bos taurus, Homo sapiens
Manually annotated by BRENDA team
Rekawiecki, R.; Nomik, M.; Kotwica, J.
Stimulatory effect of LH, PGE2 and progesterone on StAR protein, cytochrome P450 cholesterol side chain cleavage and 3beta hydroxysteroid dehydrogenase gene expression in bovine luteal cells
Prostaglandins other Lipid Mediat.
78
169-184
2005
Bos taurus
Manually annotated by BRENDA team
Chiu, C.H.; Wei, H.W.; Wu, L.S.
Generation and utilization of P450 cholesterol side-chain cleavage enzyme and 3beta-hydroxysteroid dehydrogenase antibodies for universal detection
J. Immunoassay Immunochem.
29
152-160
2008
Bos taurus, Sus scrofa, Capra aegagrus
Manually annotated by BRENDA team
Nguyen, M.N.; Slominski, A.; Li, W.; Ng, Y.R.; Tuckey, R.C.
Metabolism of vitamin d2 to 17,20,24-trihydroxyvitamin d2 by cytochrome p450scc (CYP11A1)
Drug Metab. Dispos.
37
761-767
2009
Bos taurus
Manually annotated by BRENDA team
Tuckey, R.C.; Li, W.; Zjawiony, J.K.; Zmijewski, M.A.; Nguyen, M.N.; Sweatman, T.; Miller, D.; Slominski, A.
Pathways and products for the metabolism of vitamin D3 by cytochrome P450scc
FEBS J.
275
2585-2596
2008
Bos taurus
Manually annotated by BRENDA team
Spivak, S.; Berdichevets, I.; Iarmolinskiǐ, D.; Maneshina, T.; Shpakovskiǐ, G.; Kartel, N.
Construction and characteristics of transgenic tobacco Nicotiana tabacum L. plants expressing CYP11A1 cDNA encoding cytochrome P450scc
Genetika
45
1217-1224
2009
Bos taurus
Manually annotated by BRENDA team
Tuckey, R.C.; Nguyen, M.N.; Slominski, A.
Kinetics of vitamin D3 metabolism by cytochrome P450scc (CYP11A1) in phospholipid vesicles and cyclodextrin
Int. J. Biochem. Cell Biol.
40
2619-2626
2008
Bos taurus
Manually annotated by BRENDA team
Pikuleva, I.A.; Mast, N.; Liao, W.L.; Turko, I.V.
Studies of membrane topology of mitochondrial cholesterol hydroxylases CYPs 27A1 and 11A1
Lipids
43
1127-1132
2008
Bos taurus
Manually annotated by BRENDA team
Irving-Rodgers, H.F.; Harland, M.L.; Sullivan, T.R.; Rodgers, R.J.
Studies of granulosa cell maturation in dominant and subordinate bovine follicles: novel extracellular matrix focimatrix is co-ordinately regulated with cholesterol side-chain cleavage CYP11A1
Reproduction
137
825-834
2009
Bos taurus
Manually annotated by BRENDA team
Mast, N.; Annalora, A.; Lodowski, D.; Palczewski, K.; Stout, C.; Pikuleva, I.
Structural basis for three-step sequential catalysis by the cholesterol side chain cleavage enzyme CYP11A1
J. Biol. Chem.
286
5607-5613
2011
Bos taurus (P00189)
Manually annotated by BRENDA team
Kubeil, C.; Yeung, J.; Tuckey, R.; Rodgers, R.; Martin, L.
Membrane-mediated protein-protein interactions of cholesterol side-chain cleavage cytochrome P450 with its associated electron transport proteins
ChemPlusChem
81
995-1002
2016
Bos taurus (P00189)
-
Manually annotated by BRENDA team
Efimova, V.S.; Isaeva, L.V.; Makeeva, D.S.; Rubtsov, M.A.; Novikova, L.A.
Expression of cholesterol hydroxylase/lyase system proteins in yeast S. cerevisiae cells as a self-processing polyprotein
Mol. Biotechnol.
59
394-406
2017
Bos taurus (P00189), Bos taurus
Manually annotated by BRENDA team