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Cholesterol side-chain cleavage enzyme

The cholesterol side-chain cleavage enzyme, also known as CYP11A1 or P450scc, is a mitochondrial cytochrome P450 enzyme encoded by the CYP11A1 gene on human chromosome 15q24.1 that catalyzes the conversion of cholesterol to pregnenolone, the initial and rate-limiting step in steroid hormone biosynthesis.^[1]^[2] This enzymatic reaction occurs in the inner mitochondrial membrane of steroidogenic cells and proceeds through three sequential monooxygenation steps—hydroxylations at C22 and C20, followed by cleavage of the C20-C22 bond—to produce pregnenolone and isocaproaldehyde.^[3]^[4] CYP11A1 is predominantly expressed in tissues involved in steroid production, including the adrenal cortex, gonads (testes and ovaries), and placenta, where it initiates the synthesis of all classes of steroid hormones such as glucocorticoids, mineralocorticoids, and sex steroids.^[2]^[5] The enzyme's activity is tightly regulated by the steroidogenic acute regulatory protein (StAR), which facilitates cholesterol transport to the mitochondrial inner membrane, and by cAMP-dependent signaling pathways activated by hormones like adrenocorticotropic hormone (ACTH) and luteinizing hormone (LH).^[2]^[5] Structural studies, including X-ray crystal structures of human and bovine CYP11A1 in complex with cholesterol and reaction intermediates like 22-hydroxycholesterol, reveal a canonical cytochrome P450 fold comprising 12 α-helices (A–L), β-sheets, and a heme-containing active site cavity that accommodates the substrate's hydrophobic side chain.^[3]^[6]^[7] Deficiencies in CYP11A1, resulting from mutations in the CYP11A1 gene, cause a rare form of congenital adrenal hyperplasia characterized by adrenal insufficiency, impaired steroidogenesis, and sex reversal in genetic males due to insufficient cortisol, aldosterone, and androgen production. In recent years, selective inhibitors of CYP11A1, such as MK-5684, have entered clinical trials as potential therapies for castration-resistant prostate cancer by blocking steroid hormone production.^[8]^[9]^[10] Beyond its canonical role, CYP11A1 exhibits broader substrate specificity, metabolizing compounds like 7-dehydrocholesterol to 7-dehydropregnenolone and vitamin D3 to 20S-hydroxyvitamin D3, a derivative with demonstrated antiproliferative and prodifferentiation effects in skin keratinocytes.^[2] These novel activities suggest potential non-steroidogenic functions in cholesterol homeostasis and vitamin D signaling, underscoring the enzyme's physiological versatility.^[2]

Nomenclature and Genetics

Nomenclature

The cholesterol side-chain cleavage enzyme is officially classified under the Enzyme Commission number EC 1.14.15.6, with the systematic name cholesterol monooxygenase (side-chain-cleaving).^[11] This nomenclature reflects its role as an oxidoreductase that catalyzes the monooxygenation and subsequent cleavage of the cholesterol side chain, utilizing reduced adrenodoxin and molecular oxygen as cofactors. Commonly used synonyms for the enzyme include CYP11A1, reflecting its identity as a member of the cytochrome P450 superfamily; P450scc, denoting its specific side-chain cleavage activity; cholesterol desmolase; and simply side-chain cleavage enzyme (SCC). These terms are widely adopted in biochemical literature and databases to describe the same protein encoded by the human CYP11A1 gene.^[12] The enzyme's classification as a cytochrome P450 places it within the CYP11 family (involved in steroid metabolism), subfamily A, and polypeptide 1, highlighting its evolutionary and functional ties to other heme-containing monooxygenases essential for endogenous substrate transformations.^[12] Historically, the enzyme's activity was first characterized in the 1960s as the rate-limiting step in steroid biosynthesis, with seminal identification of its mitochondrial localization and catalytic function reported in 1966 by Simpson and Boyd using adrenal and placental tissues.^[13] This discovery built on earlier 1960s studies linking adrenocorticotropic hormone (ACTH) stimulation to protein synthesis requirements for steroid production, establishing the enzyme's central nomenclature in endocrinology.^[13]

Gene Structure

The CYP11A1 gene is located on the long arm of human chromosome 15 at the cytogenetic band 15q24.1, spanning approximately 29.9 kb on the reverse strand from positions 74,337,746 to 74,367,680.^[14]^[15] The gene consists of 9 exons and 8 introns, with the first intron being notably large at about 13.8 kb and dominated by repetitive elements.^[16]^[15] The promoter region of CYP11A1 features a TATA box and binding sites for key transcription factors, including a proximal steroidogenic factor-1 (SF-1) site at approximately -40 bp relative to the transcription start site and an upstream SF-1 site around -1,600 bp.^[17] These SF-1 binding sites are essential for tissue-specific expression, with the proximal site contributing to basal promoter activity in steroidogenic tissues.^[17] Additional regulatory elements in the promoter include Sp1 binding sites and a cAMP-response sequence upstream.^[15] Several genetic variants have been identified in the CYP11A1 gene, particularly in the promoter region, that can influence expression levels. For instance, the polymorphism rs4887139 (C/T) at -2,228 bp in the promoter has been associated with altered transcriptional activity, with the C allele linked to higher expression in certain haplotypes.^[15] In population studies, the minor allele frequency of rs4887139 is approximately 0.27 in East Asian cohorts and varies up to 0.45 in European populations based on HapMap data.^[15] Another promoter variant, the microsatellite D15S520 (TAAAA)n repeat at -487 bp, shows an 8-repeat allele frequency of about 0.086 in Chinese populations, and haplotypes incorporating this repeat have been shown to increase gene expression up to twofold in lymphoblastoid cell lines.^[15] The CYP11A1 gene exhibits strong evolutionary conservation across vertebrates, with orthologs identified in over 190 species ranging from mammals (e.g., Mus musculus, Bos taurus) to fish (e.g., Danio rerio, which has duplicated cyp11a1 and cyp11a2 genes).^[14] This conservation underscores its fundamental role in steroidogenesis, with the core gene structure and promoter elements preserved from jawed vertebrates to mammals.^[18]

Protein Structure and Localization

Protein Structure

The cholesterol side-chain cleavage enzyme, also known as CYP11A1, is synthesized as a precursor protein that undergoes processing to yield the mature form with an approximate molecular mass of 55 kDa. This maturation involves the cleavage and removal of an N-terminal mitochondrial import signal sequence of 39 amino acids, which directs the protein to the mitochondrial matrix and inner membrane. The resulting mature protein consists of 482 amino acids and adopts a structure optimized for its role in steroid biosynthesis within the mitochondrial environment.^[19]^[7] CYP11A1 exhibits the characteristic domain organization of mitochondrial cytochrome P450 enzymes, featuring an N-terminal helical domain followed by a core beta-sheet cytochrome P450 domain. The helical domain includes alpha-helices A through L, with additional unique extensions such as helices A' and K'', which contribute to membrane association and substrate access. The beta-sheet domain houses the heme prosthetic group, coordinated by a conserved cysteine residue within the heme-binding motif FxxGxRxCxG (specifically, Phe435-Gly438-Arg441-Cys444-Gly445 in human CYP11A1), essential for oxygen activation and catalysis. This motif is part of a broader conserved region that stabilizes the heme and facilitates electron transfer from redox partners. Unlike some eukaryotic proteins, CYP11A1 lacks N-glycosylation sites, reflecting its synthesis and maturation outside the endoplasmic reticulum-Golgi pathway.^[4]^[20]^[21] High-resolution crystal structures, such as that deposited in PDB entry 3NA0 (resolved at 2.50 Å), provide detailed insights into the enzyme's architecture, capturing CYP11A1 in complex with adrenodoxin and the intermediate 20,22-dihydroxycholesterol. These structures reveal a canonical P450 fold comprising 12 alpha-helices, four beta-sheets, and interconnecting loops, forming a triangular prism-shaped molecule with a buried heme and an accessible substrate-binding pocket. The active site pocket, lined by hydrophobic residues such as Ile99, Leu104, and Phe285, accommodates the cholesterol molecule in an extended conformation, positioning its side chain near the heme iron for sequential hydroxylations. Recent studies, including NMR analysis of CYP11A1–adrenodoxin interactions and crystal structures of ancestral forms (e.g., PDB 9NJW), further elucidate evolutionary adaptations and redox partner binding. In solution, CYP11A1 predominantly exists as a monomer, as evidenced by size-exclusion chromatography and spectroscopic analyses; however, atomic force microscopy and membrane reconstitution studies suggest potential dimerization in the lipid bilayer, possibly mediated by the F-G loop region for enhanced stability or function.^[22]^[23]^[24]^[25]^[26]

Tissue and Cellular Localization

The cholesterol side-chain cleavage enzyme, encoded by the CYP11A1 gene, is primarily expressed in steroidogenic tissues, including the adrenal cortex, gonads, and placenta. In the adrenal gland, it is highly enriched in the zona fasciculata and zona reticularis of the cortex, where it supports glucocorticoid and mineralocorticoid production. Within the gonads, expression is prominent in Leydig cells of the testis and granulosa-lutein cells of the ovary, facilitating androgen and estrogen biosynthesis, respectively. In the placenta, CYP11A1 is localized to syncytiotrophoblast cells, contributing to progesterone synthesis essential for pregnancy maintenance.^[27]^[28]^[29] At the subcellular level, CYP11A1 is anchored to the inner mitochondrial membrane in a monotopic orientation, with the majority of the protein, including its active site, facing the mitochondrial matrix to enable efficient catalysis of cholesterol side-chain cleavage. This positioning allows direct access to cholesterol substrates delivered to the matrix side of the membrane. Quantitative expression data from the GTEx database reveal very high levels in the adrenal gland (median TPM ~1056) and lower but significant levels in testis (median TPM ~22), with negligible levels in non-steroidogenic tissues such as liver or brain. Placenta shows moderate expression based on other datasets. Fetal testis exhibits elevated expression (median TPM ~20.5).^[30]^[31]^[32]^[33] Developmentally, CYP11A1 expression in the human fetal adrenal gland is upregulated by approximately week 8 of gestation, coinciding with the onset of cortisol production in response to adrenocorticotropic hormone (ACTH), which supports fetal organ maturation. This early activation marks the beginning of functional steroidogenesis in the provisional zone of the fetal adrenal cortex. In non-mammalian species, such as teleost fish, CYP11A1 expression exhibits broader patterns beyond classical steroidogenic tissues; for instance, in zebrafish, duplicate cyp11a paralogs (cyp11a1 and cyp11a2) are expressed not only in gonads and interrenal (adrenal equivalent) tissue but also in the brain, enabling extra-adrenal steroid production for neurosteroid roles. Similarly, in species like the olive flounder, cyp11a1 is detected in head kidney and additional peripheral sites, reflecting adaptations for diverse steroid functions in aquatic environments.^[34]^[35]^[36]^[37]

Biochemical Function

Mechanism of Action

The cholesterol side-chain cleavage enzyme, also known as CYP11A1, catalyzes the conversion of cholesterol to pregnenolone through three sequential monooxygenation reactions occurring at the mitochondrial inner membrane, which facilitates access to the substrate cholesterol.^[38] The first step involves the 22R-hydroxylation of cholesterol to form (22R)-hydroxycholesterol, mediated by the enzyme's heme iron center activating molecular oxygen. This is followed by a second monooxygenation at the 20α position (20R configuration), yielding (20R,22R)-dihydroxycholesterol as the key intermediate. The third and final step entails oxidative cleavage of the C20-C22 bond in (20R,22R)-dihydroxycholesterol, producing pregnenolone and isocaproaldehyde (4-methylpentanal), with involvement of radical intermediates during the bond scission facilitated by the ferryl-oxo species (Compound I).^[39] Each monooxygenation step requires molecular oxygen (O₂) and reducing equivalents from NADPH, delivered via the electron transport chain involving adrenodoxin reductase and adrenodoxin (Adx), a ferredoxin that shuttles electrons to the P450 heme.^[38] The overall reaction stoichiometry is:

\text{Cholesterol} + 3\, \text{O}_2 + 3\, \text{NADPH} + 3\, \text{H}^+ \rightarrow \text{[Pregnenolone](/page/Pregnenolone)} + \text{isocaproaldehyde} + 3\, \text{H}_2\text{O} + 3\, \text{NADP}^+

This six-electron oxidation process is highly processive, with intermediates remaining bound to the enzyme active site to minimize dissociation.^[39] Kinetic studies indicate a Michaelis constant (K_m) for cholesterol of approximately 1-5 μM under reconstituted conditions, reflecting efficient substrate binding despite cholesterol's hydrophobicity.^[38] The turnover rate (k_cat) ranges from 1-5 min⁻¹, limiting the overall rate of steroidogenesis.^[39] As a heme-containing cytochrome P450, CYP11A1 is inhibited by carbon monoxide, which binds to the ferrous heme and prevents O₂ activation, underscoring the enzyme's dependence on the porphyrin cofactor for catalysis.^[38]

Role in Steroidogenesis

The cholesterol side-chain cleavage enzyme, also known as CYP11A1 or P450scc, catalyzes the conversion of cholesterol to pregnenolone through a series of hydroxylations and side-chain cleavage, marking the committed and rate-limiting initial step in the biosynthesis of all steroid hormones.^[8] This process occurs on the inner mitochondrial membrane of steroidogenic cells and serves as the foundational gateway for producing glucocorticoids, mineralocorticoids, and sex steroids across diverse endocrine tissues, including the adrenal cortex, gonads, and placenta.^[5] By transforming cholesterol—a ubiquitous lipid precursor—into pregnenolone, CYP11A1 enables the divergence of steroid pathways that are critical for stress response, electrolyte balance, and reproductive function.^[40] In the broader steroidogenic cascade, pregnenolone generated by CYP11A1 is rapidly metabolized by downstream enzymes to yield tissue-specific hormones. For instance, 3β-hydroxysteroid dehydrogenase (3β-HSD) converts pregnenolone to progesterone in the mitochondria, providing a substrate for further processing, while CYP17A1 performs 17α-hydroxylation and 17,20-lyase activities in the endoplasmic reticulum to direct synthesis toward androgens or estrogens depending on the cellular context.^[5] In the adrenal glands, this integration under adrenocorticotropic hormone (ACTH) stimulation amplifies steroid output, with CYP11A1 controlling the majority of glucocorticoid production—such as cortisol in the zona fasciculata—by facilitating cholesterol mobilization via the steroidogenic acute regulatory protein (StAR).^[41] This coordinated enzymatic progression ensures efficient, localized hormone production tailored to physiological demands, such as ACTH-driven responses that enhance adrenal capacity for stress adaptation.^[42] Beyond its canonical function, CYP11A1 supports non-steroidogenic roles in extra-adrenal sites like the skin, where it hydroxylates vitamin D3 at the 20-position to generate bioactive precursors such as 20-hydroxyvitamin D3 and 20,23-dihydroxyvitamin D3, which promote keratinocyte differentiation, inhibit proliferation, and provide photoprotection against UVB damage without significant calcemic effects.^[2] These metabolites, produced in epidermal keratinocytes, contribute to cutaneous barrier integrity and immune modulation, highlighting CYP11A1's versatility in peripheral tissues.^[43] Evolutionarily, CYP11A1 represents a conserved vertebrate innovation as the primary gateway enzyme in steroidogenesis, with ancestral reconstructions revealing high structural and functional similarity in catalytic domains across species from early chordates to mammals, underscoring its essential role in the emergence of complex endocrine systems.^[44]

Regulation

Transcriptional Regulation

The transcription of the CYP11A1 gene, encoding the cholesterol side-chain cleavage enzyme, is primarily regulated by the orphan nuclear receptor steroidogenic factor-1 (SF-1, also known as NR5A1), which binds to specific sites in the promoter region to drive basal expression and enhance cAMP-induced transcription in steroidogenic tissues such as the adrenal cortex and gonads.^[45] SF-1 interacts with coactivators like CBP/p300 to facilitate chromatin remodeling and gene activation, ensuring tissue-specific expression essential for steroidogenesis.^[46] Additionally, the cAMP-responsive element-binding protein (CREB) plays a critical role in the acute hormonal response by binding to cAMP response elements (CREs) in the promoter, where phosphorylation by protein kinase A (PKA) triggers rapid transcriptional upregulation following cAMP elevation.^[46] Tissue-specific enhancers further refine CYP11A1 expression; for instance, GATA transcription factors, particularly GATA-6, cooperate with SF-1 in gonadal cells to potentiate promoter activity, while in the placenta, the related nuclear receptor NR5A2 (also known as LRH-1) binds similar response elements to drive expression independently of SF-1.^[46]^[47] Hormonally, adrenocorticotropic hormone (ACTH) in the adrenal cortex and luteinizing hormone (LH)/follicle-stimulating hormone (FSH) in the gonads stimulate CYP11A1 transcription via the cAMP/PKA pathway, leading to CREB phosphorylation and SF-1 activation for increased steroid production.^[45] This positive regulation is counterbalanced by negative feedback, where glucocorticoids bind the glucocorticoid receptor (GR) in the hypothalamus and pituitary to repress corticotropin-releasing hormone (CRH) and ACTH secretion, thereby indirectly reducing CYP11A1 expression in the adrenal gland.^[48] Epigenetic modifications also govern CYP11A1 accessibility; in expressing steroidogenic cells, histone acetylation (e.g., on H3 and H4) at the promoter loosens chromatin structure to promote transcription factor binding, whereas in non-expressing tissues, hypermethylation of CpG islands in the promoter region silences the gene by inhibiting SF-1 recruitment and maintaining a repressive chromatin state.^[49] During embryonic development, retinoic acid signaling activates CYP11A1 expression in fetal steroidogenic cells, such as Leydig precursors, by enhancing SF-1-dependent transcription in combination with cAMP stimuli, contributing to the timely differentiation of adrenal and gonadal tissues.^[50]

Post-translational Regulation

The activity of cholesterol side-chain cleavage enzyme (CYP11A1) is critically dependent on the steroidogenic acute regulatory protein (StAR), which facilitates cholesterol transport from the outer mitochondrial membrane to the inner membrane where CYP11A1 is localized. This cholesterol import step is rate-limiting during acute hormonal stimulation of steroidogenesis, as StAR enhances substrate availability without altering CYP11A1's intrinsic catalytic rate.^[51]^[52] Under basal conditions, CYP11A1 activity is substrate-limited, but StAR activation rapidly increases pregnenolone production by promoting cholesterol delivery to the enzyme's active site.^[53] Post-translational phosphorylation modulates both StAR and CYP11A1 function. For StAR, protein kinase A (PKA) phosphorylates serine 194 (in mice) or serine 195 (in humans), approximately doubling its cholesterol-transfer efficiency and thereby amplifying CYP11A1 activity; conversely, dephosphorylation by protein phosphatase 2A (PP2A) diminishes this effect, reducing steroidogenic output.^[13] Directly on CYP11A1, PKA and protein kinase C (PKC) phosphorylate serine and threonine residues, enhancing enzymatic activity by improving electron transfer efficiency from its redox partner adrenodoxin and increasing overall pregnenolone synthesis rates.^[54]^[55] Redox regulation of CYP11A1 occurs primarily through its interaction with adrenodoxin, an iron-sulfur protein that delivers electrons from NADPH-adrenodoxin reductase; variations in adrenodoxin availability directly influence CYP11A1's reduction potential and catalytic turnover.^[56] Superoxide modulation further impacts this process, as elevated reactive oxygen species in steroidogenic mitochondria can impair adrenodoxin function and CYP11A1 activity, while superoxide dismutase protects against such inhibition to maintain steroid output.^[57] Substrate binding to CYP11A1 induces allosteric conformational changes that strengthen its interaction with adrenodoxin, optimizing electron delivery for the multi-step cholesterol cleavage reaction.^[25] Protein-protein interactions also stabilize CYP11A1 function; for instance, adrenodoxin binding supports electron transfer.^[58]

Clinical and Pathological Aspects

Associated Diseases

Mutations in the CYP11A1 gene, encoding the cholesterol side-chain cleavage enzyme (P450scc), cause a rare autosomal recessive form of congenital adrenal hyperplasia characterized by primary adrenal insufficiency and impaired steroidogenesis, clinically resembling but distinct from classic lipoid congenital adrenal hyperplasia due to StAR deficiency.^[59] This disorder, known as adrenal insufficiency, congenital, with 46,XY sex reversal, type 1 (AICSR1; OMIM 613743), results from defective conversion of cholesterol to pregnenolone, leading to deficiencies in glucocorticoids, mineralocorticoids, and sex steroids.^[59] Affected individuals typically present with salt-wasting adrenal crisis in infancy or early childhood, hyponatremia, hyperkalemia, and hypoglycemia, often requiring lifelong hormone replacement therapy.^[60] In 46,XY individuals, the steroid deficiency disrupts gonadal development, causing a spectrum of disorders of sex development ranging from female external genitalia to hypospadias and partial virilization, while 46,XX individuals may exhibit normal female genitalia but adrenal insufficiency.^[61] Unlike StAR deficiency, CYP11A1 mutations do not typically cause massive lipid-laden adrenal enlargement, though cholesterol accumulation in steroidogenic tissues can occur due to blocked pregnenolone synthesis.^[59] Over 50 cases have been reported worldwide, with identified mutations including homozygous or compound heterozygous variants such as p.Leu222Pro, p.Arg405Pro, and p.Arg451Trp, which severely impair enzymatic activity.^[62]^[61] Partial deficiencies arising from milder CYP11A1 mutations manifest with delayed-onset adrenal insufficiency, often presenting beyond infancy with fatigue, poor weight gain, or stress-induced crises, and may include isolated hypogonadism without full sex reversal.^[60] For instance, compound heterozygous mutations like p.Glu314Lys and a splice-site variant can retain residual enzyme activity (10-20% of normal), leading to partial steroid production sufficient for survival but inadequate during stress, and in males, associated with micropenis, cryptorchidism, or later hypogonadotropic hypogonadism.^[63] These phenotypes highlight the enzyme's dose-dependent role in maintaining steroid homeostasis, with some patients achieving partial recovery of adrenal function under glucocorticoid therapy but requiring monitoring for gonadal defects.^[64] Promoter polymorphisms in CYP11A1, such as the (tttta)n repeat in the 5' untranslated region, have been associated with polycystic ovary syndrome (PCOS) by enhancing transcriptional activity and increasing ovarian androgen production.^[65] Specifically, shorter repeat alleles (e.g., 4 or 6 repeats) correlate with elevated testosterone levels and hyperandrogenemia in PCOS patients and their unaffected relatives, potentially contributing to ovarian dysfunction through upregulated pregnenolone synthesis as the first step in theca cell steroidogenesis.01468-4/abstract) These genetic variants increase PCOS susceptibility, particularly in populations with higher allele frequencies, though environmental factors also influence penetrance.^[66] CYP11A1 deficiency is extremely rare, with a prevalence estimated at less than 1 in 1,000,000 births, though higher incidence occurs in consanguineous populations due to its autosomal recessive inheritance, facilitating prenatal diagnosis via genetic testing of amniotic fluid for pathogenic variants.^[67] In animal models, homozygous Cyp11a1 knockout mice are viable at birth but exhibit neonatal lethality within days due to profound steroid deficiency, manifesting as growth retardation, dehydration, hypo- and hyperglycemia, and placental insufficiency contributing to impaired fetal adrenal and gonadal development.^[68] These mice lack all steroid hormones, underscoring the enzyme's essential role in embryonic viability and highlighting the reliance on maternal steroids for gestation.^[68]

Inhibitors and Therapeutics

Aminoglutethimide acts as a competitive inhibitor of cholesterol side-chain cleavage enzyme (CYP11A1) by binding to the heme iron in the active site, thereby preventing cholesterol substrate access and blocking the initial step of steroidogenesis.^[69] This inhibition reduces adrenal steroid production, making it useful in treating Cushing's syndrome by suppressing excess cortisol and in advanced breast cancer by limiting estrogen synthesis from adrenal precursors.^[70] Historically, aminoglutethimide combined with glucocorticoids achieved medical adrenalectomy, effectively ablating adrenal function in metastatic breast and prostate cancers to deprive tumors of steroid hormones.^[71] Azole antifungals such as ketoconazole and metyrapone serve as inhibitors of CYP11A1, with ketoconazole competitively coordinating the heme iron to disrupt enzyme activity at concentrations achieving over 65% inhibition at 10 μM.^[69] Metyrapone similarly targets CYP11A1 alongside other steroidogenic enzymes, exhibiting potency in the micromolar range (IC50 values approximately 1-10 μM for related P450 inhibitions).^[72] These agents are employed as adjunct therapies in prostate cancer for androgen deprivation by curtailing steroid precursor availability and in ectopic ACTH syndrome to control hypercortisolism, often normalizing urinary free cortisol in 30-80% of Cushing's disease cases at doses of 600-1200 mg/day for ketoconazole.^[72] Emerging selective CYP11A1 inhibitors, such as tetrandrine, covalently bind to Cys423 in the enzyme's active site, destabilizing heme and inhibiting aldosterone synthesis with an IC50 of 9.024 μM; structure-activity relationships highlight the necessity of this cysteine residue for binding efficacy, as its mutation abolishes inhibition.^[73] These compounds show promise in treating hyperaldosteronism by reducing serum aldosterone levels and exerting antihypertensive effects in animal models without broadly disrupting other steroid pathways.^[73] Similarly, investigational agents like ODM-208 demonstrate antitumor activity in metastatic castration-resistant prostate cancer, achieving PSA50 responses in over 50% of patients with androgen receptor mutations.^[74] Common side effects of CYP11A1 inhibitors include hypoaldosteronism manifesting as adrenal insufficiency (reported in 13-36% of patients), fatigue (affecting 30-38%), and potential gynecomastia due to altered steroid balances; clinical management involves monitoring serum steroid levels and providing glucocorticoid replacement as needed.^[74]

References

[1]
CYP11A1 cytochrome P450 family 11 subfamily A member 1 ... - NCBI
Sep 5, 2025 · CYP11A1 encodes a cytochrome P450 enzyme that converts cholesterol to pregnenolone, the first step in steroid hormone synthesis.
[2]
Novel activities of CYP11A1 and their potential physiological ... - NIH
CYP11A1, found only in vertebrates, catalyzes the first step of steroidogenesis where cholesterol is converted to pregnenolone.
[3]
Structural basis for three-step sequential catalysis by the cholesterol ...
Feb 18, 2011 · We present the 2.5-Å crystal structure of CYP11A1 in complex with the first reaction intermediate, 22HC. The active site cavity in CYP11A1 represents a long ...
[4]
Structural Basis for Three-step Sequential Catalysis by the ... - NIH
Herein, we present the 2.5-Å crystal structure of CYP11A1 in complex with the first reaction intermediate, 22HC. The active site cavity in CYP11A1 represents a ...
[5]
Early steps in steroidogenesis: intracellular cholesterol trafficking - NIH
Cytochrome P450scc (CYP11A1) then initiates steroidogenesis by converting cholesterol to pregnenolone on the inner mitochondrial membrane (IMM).
[6]
3N9Y: Crystal structure of human CYP11A1 in complex with ... - NCBI
May 31, 2010 · Molecular Graphic ? 3N9Y: Crystal structure of human CYP11A1 in complex with cholesterol. 3D view. full-featured 3D viewer. Download Cn3D.
[7]
The A'-helix of CYP11A1 remodels mitochondrial cristae - PMC
Aug 18, 2022 · CYP11A1 participates in the formation of tubulovesicular cristae in the mitochondria of steroidogenic cells. Its A'-helix is sufficient for the formation of ...Fig. 3 · Cyp11a1 Does Not Change... · Hsp60 Together With Cyp11a1...<|control11|><|separator|>
[8]
Cholesterol Side-Chain Cleavage Enzyme (SCC) Deficiency - PMC
Cholesterol side-chain cleavage enzyme (SCC) catalyzes the conversion of cholesterol to pregnenolone in steroidogenic cells, which is the first step in the ...
[9]
Partial Defect in the Cholesterol Side-Chain Cleavage Enzyme ... - NIH
The cholesterol side-chain cleavage enzyme (P450scc), encoded by the CYP11A1 gene, converts cholesterol to pregnenolone to initiate steroidogenesis. Genetic ...
[10]
EC 1.14.15.6 - IUBMB Nomenclature
EC 1.14.15.6. Accepted name: cholesterol monooxygenase (side-chain-cleaving). Reaction: cholesterol + 6 reduced adrenodoxin + 3 O2 + ...
[11]
CYTOCHROME P450, SUBFAMILY XIA, POLYPEPTIDE 1; CYP11A1
The cholesterol side-chain cleavage enzyme (P450scc; EC 1.14.15.6), encoded by the CYP11A1 gene, initiates steroidogenesis by converting cholesterol to ...Missing: systematic | Show results with:systematic
[12]
Thirty years of StAR gazing. Expanding the universe of the ...
This brief overview provides a timeline of events in four converging areas of study: mechanisms of the acute regulation of steroidogenesis, leading to StAR; ...
[13]
Gene: CYP11A1 (ENSG00000140459) - Summary - Homo_sapiens
Chromosome 15: 74,337,746-74,367,680 reverse strand. This gene has 13 transcripts (splice variants), 1 gene allele, 193 orthologues, 2 paralogues and is ...
[14]
Haplotype Analysis of CYP11A1 Identifies Promoter Variants ...
Here we conduct a systematic investigation of the role of genetic variation of the CYP11A1 gene in breast cancer risk in a study of 1193 breast cancer cases.Missing: rs2279356 | Show results with:rs2279356
[15]
Homozygous Disruption of P450 Side-Chain Cleavage (CYP11A1 ...
P450scc is encoded by the CYP11A1 gene, which is 29 kb long and consists of nine exons and eight introns. The resulting protein is 521 amino acids in length and ...
[16]
Functions of the upstream and proximal steroidogenic factor 1 (SF-1)
Two functional SF-1-binding sites, P and U, located at -40 and -1,600 regions of the CYP11A1 gene, have been identified, but their exact functions with respect ...
[17]
Evolution of vertebrate Cyp11a orthologs. Maximum likelihood...
... cyp11a1 is maternally expressed and cyp11a2 is considered the ortholog of Cyp11a1 in mammals. ... conserved among non-mammalian vertebrates, especially in fish.
[18]
https://www.researchgate.net/figure/Evolution-of-vertebrate-Cyp11a-orthologs-Maximum-likelihood-phylogenetic-analyses-of_fig2_236741092
[19]
Computational Identification of the Paralogs and Orthologs of ...
Jun 28, 2016 · We have identified at least 3 well conserved motifs in human CYPs: “FXXGXRXCXG” located in the heme-binding domain, “AGXDTT”, and “EXXR” located ...
[20]
p05108 · cp11a_human - UniProt
A cytochrome P450 monooxygenase that catalyzes the side-chain hydroxylation and cleavage of cholesterol to pregnenolone, the precursor of most steroid hormones.
[21]
RCSB PDB - 3NA0: Crystal structure of human CYP11A1 in complex with 20,22-dihydroxycholesterol
### Summary of Crystal Structure of CYP11A1 (PDB ID: 3NA0)
[22]
https://www.rcsb.org/structure/3NA0
[23]
Atomic Force Microscopy Study of Protein–Protein Interactions in the ...
UV/VIS spectra of monomeric or oligomeric proteins revealed no significant changes (data not shown). UV/VIS spectra of CYP11A1 displayed a pronounced peak at ...
[24]
The Steroidogenic Enzyme Cyp11a1 is Essential for Development of ...
Cyp11a1 is expressed primarily in the cortex of the adrenal gland, but testis, ovary, placenta, thymus, and intestine also express Cyp11a1 (6, 9).
[25]
CYP11A1 protein expression summary - The Human Protein Atlas
The categories include: tissue enriched, group enriched, tissue enhanced, low tissue specificity and not detected. Tissue enriched (Adrenal gland).Missing: mature size
[26]
Tissue-Specific, Hormonal, and Developmental Regulation of SCC ...
This expression pattern correlates well with the function of steroids in ovarian cycles. In the testis, CYP11A1 is expressed in the Leydig cells (17).β-Galactosidase Assays · Results · Analysis Of Cyp11a1 Promoter...
[27]
The A'-helix of CYP11A1 remodels mitochondrial cristae
Aug 18, 2022 · The mature CYP11A1 is composed of 12 helices (helix A to L), four β sheets, and the loops between these structures [5]. CYP11A1 is partially ...Missing: organization | Show results with:organization
[28]
The F–G loop region of cytochrome P450scc (CYP11A1) interacts ...
We conclude that CYP11A1 has a monotopic association with the membrane that is mediated, at least in part, by the F–G loop region.Missing: organization | Show results with:organization<|control11|><|separator|>
[29]
CYP11A1 Gene - GeneCards | CP11A Protein
This gene encodes a member of the cytochrome P450 superfamily of enzymes. The cytochrome P450 proteins are monooxygenases which catalyze many reactions ...
[30]
The human fetal adrenal produces cortisol but no detectable ... - NIH
Feb 12, 2018 · The adrenals secrete cortisol in response to adrenocorticotropic hormone (ACTH) as early as week 8 of gestation [1], although the main steroids ...
[31]
Development and function of the fetal adrenal
Oct 18, 2022 · The earliest significant function of the fetal adrenals appears to be the timely production of cortisol between 8 and 10 weeks-post-conception ( ...
[32]
Identification and developmental expression of the full complement ...
Nov 18, 2010 · Here we identify and annotate the full suite of CYP genes in zebrafish, compare these to the human CYP gene complement, and determine the expression of CYP ...
[33]
The Brain of Teleost Fish, a Source, and a Target of Sexual Steroids
This article aims at reviewing the available information regarding expression and/or activity of the main steroidogenic enzymes in the brain of fish.
[34]
Rapid kinetic methods to dissect steroidogenic cytochrome P450 ...
The cholesterol side-chain cleavage enzyme (CYP11A1) catalyzes the conversion of cholesterol to pregnenolone in three steps, via the sequential intermediates ...
[35]
The multistep oxidation of cholesterol to pregnenolone by human ...
Cytochrome P450 (P450, CYP) 11A1 is the classical cholesterol side chain cleavage enzyme (P450scc) that removes six carbons of the side chain, the first and ...<|control11|><|separator|>
[36]
Cholesterol Side-Chain Cleavage Enzyme - ScienceDirect.com
The primary enzyme that catalyzes the cleavage of the side chain of cholesterol to initiate the process of steroidogenesis is P450scc that produces as its ...
[37]
ACTH Regulation of Adrenal SR-B1 - Frontiers
ACTH Stimulation Regulates the Expression of SR-B1. Initial studies established a functional correlation between SR-B1 expression, HDL-CE uptake, and the ...
[38]
Impact of ACTH Signaling on Transcriptional Regulation of ... - NIH
Mar 29, 2016 · The trophic peptide hormone adrenocorticotropic (ACTH) stimulates steroid hormone biosynthesis evoking both a rapid, acute response and a long- ...
[39]
The significance of CYP11A1 expression in skin physiology ... - NIH
Vitamin D analogs 17,20S(OH)2pD and 17,20R(OH)2pD are noncalcemic and exhibit antifibrotic activity, J Invest Dermatol. 131, 1167–9. [DOI] [PMC free article] ...
[40]
Resurrection and characterization of ancestral CYP11A1 enzymes
Jun 6, 2021 · Mitochondrial P450s are localized on the matrix side of the inner mitochondrial membrane and are primarily involved in biosynthesis of steroid ...
[41]
Cyclic AMP Stimulates SF-1-Dependent CYP11A1 Expression ... - NIH
Steroidogenic factor 1 (SF-1) (also called Ad4BP or NR5A1) is a ... In this study, we examined the mechanism underlying SF-1-regulated CYP11A1 expression.
[42]
Impact of ACTH Signaling on Transcriptional Regulation ... - Frontiers
Transcriptional regulation of the cholesterol side chain cleavage cytochrome P450 gene (CYP11A1) revisited: binding of GATA, cyclic adenosine 3′,5 ...
[43]
Liver receptor homologue-1 is expressed in human steroidogenic tissues and activates transcription of genes encoding steroidogenic enzymes
### Summary of LRH-1 (NR5A2) Expression and Regulation of CYP11A1 in Steroidogenic Tissues (Placenta)
[44]
Role of glucocorticoid negative feedback in the regulation of HPA ...
In this review, we will discuss our knowledge of the negative feedback mechanisms that regulate basal ultradian synthesis and secretion of glucocorticoids.
[45]
Epigenetic Changes of the Cyp11a1 Promoter Region in Granulosa ...
This study investigated in vivo whether epigenetic controls including histone modifications and DNA methylation in the promoter region are associated with the ...
[46]
Steroidogenic Factor 1 Differentially Regulates Fetal and Adult ... - NIH
... Cyp11a1 was expressed only after treatment with cAMP and retinoic acid. Together, data show SF-1 is needed for proper development of fetal and adult Leydig ...
[47]
STAR/STARD1: A mitochondrial intermembrane space cholesterol ...
The import of cholesterol to the inner mitochondrial membrane by the steroidogenic acute regulatory protein (STAR/STARD1) is essential for de novo steroid ...
[48]
Mitochondrial Protein Import and the Genesis of Steroidogenic ...
Human CYP11A1 is encoded as a 521-amino acid polypeptide with a 39-amino acid mitochondrial presequence, which is cleaved upon import (DuBois et al., 1981).
[49]
STARD1 Functions in Mitochondrial Cholesterol Metabolism and ...
STARD1 moves cholesterol (CHOL) from the outer mitochondrial membrane (OMM) to the inner membrane (IMM) in steroidogenic cells.Abstract · Introduction · The Unique Role of STARD1... · Complexity of STARD1...
[50]
The Catalytic Mechanism of Steroidogenic Cytochromes P450 from ...
Experimental data indicate that PKA can phosphorylate CYP11A1 serine and threonine amino acids, enhancing as a result their enzymatic activity [106]. In ...Cyp19a1 And Cyp17a1 On The... · 6.1. Cyp19a1 Phosphorylation · 6.3. Cyp11a1 Phosphorylation
[51]
https://www.pnas.org/doi/10.1073/pnas.2508809122
[52]
Redox Partner Adrenodoxin Induces Substrate Binding to ...
Oct 6, 2025 · CYP11A1 converts cholesterol to pregnenolone in three steps, including two hydroxylation steps on the C-20 and C-22 positions, followed by a ...Missing: stereochemistry 20S-
[53]
Ablation of Steroidogenic Superoxide Dismutase 2 Increases ...
At low to modest doses, ROS play an essential role in the regulation of normal physiological functions, such as cell-cycle progression and proliferation, ...
[54]
NMR-guided identification of CYP11A1–Adrenodoxin interactions ...
Jun 26, 2025 · The crystal structure of the human CYP11A1-Adx complex positions the Adx α helix-3 residue L80 as forming a likely nonpolar interaction ...<|control11|><|separator|>
[55]
The Interaction of Bovine Adrenodoxin with CYP11A1 (Cytochrome ...
The adrenodoxin mutants forming the most stable complexes with CYP11A1 show the fastest rates of reduction and the highest rate constants for cholesterol to ...Missing: superoxide | Show results with:superoxide
[56]
613743 - ADRENAL INSUFFICIENCY, CONGENITAL, WITH 46,XY ...
P450scc deficiency is a rare disorder that can present as acute adrenal insufficiency in infancy or childhood.
[57]
Partial Defect in the Cholesterol Side-Chain Cleavage Enzyme ...
The cholesterol side-chain cleavage enzyme (P450scc), encoded by the CYP11A1 gene, converts cholesterol to pregnenolone to initiate steroidogenesis. Genetic ...Missing: translational | Show results with:translational
[58]
Primary Adrenal Insufficiency, Complete Sex Reversal, and Unique ...
Oct 12, 2024 · CYP11A1 (P450scc) deficiency is a rare and complex disorder that leads to primary adrenal insufficiency and may present with 46, XY disorders of ...
[59]
A novel homozygous mutation in CYP11A1 gene is associated with ...
A novel homozygous mutation in CYP11A1 gene is associated with late-onset adrenal insufficiency and hypospadias in a 46,XY patient.Missing: rs2279356 polymorphism promoter
[60]
A novel splice site variant in CYP11A1 in trans with the p.E314K ...
Jul 20, 2017 · CYP11A1 deficiency results in complete or partial adrenal insufficiency with a wide range of clinical manifestations. Severe or classical ...Missing: hypogonadism | Show results with:hypogonadism
[61]
Long-term outcome of partial P450 side-chain cleavage enzyme ...
The P450scc enzyme plays a key role in the initial steps of steroidogenesis by catalyzing the conversion of cholesterol to pregnenolone in steroidogenic tissues ...
[62]
CYP11A1 and CYP17 promoter polymorphisms associate ... - PubMed
Polymorphisms in promoter regions of the two key androgen-regulating genes, CYP11A1 and CYP17, were found to be significantly associated with T levels.Missing: rs2279356 | Show results with:rs2279356
[63]
CYP11A1 microsatellite (tttta)n polymorphism in PCOS women from ...
Conclusion. CYP11A1 (tttta)n repeat polymorphism appeared to be a potential molecular marker for PCOS risk in our population. Gene-gene and gene-environmental ...
[64]
46,XY difference of sex development-adrenal insufficiency due to ...
46,XY disorder of sex development-adrenal insufficiency due to CYP11A1 deficiency; XY sex reversal-adrenal failure. Prevalence: <1 / 1 000 000. Inheritance ...
[65]
Steroid Deficiency Syndromes in Mice with Targeted Disruption of ...
Cyp11a1 knockout mice manifested growth retardation, muscle atrophy, dehydration, lethargy, and anorexia. In addition, XY Cyp11a1 knockout pups exhibited ...Abstract · RESULTS · DISCUSSION · MATERIALS AND METHODS
[66]
Inhibition and stimulation of activity of purified recombinant ... - NIH
Oct 23, 2012 · In vertebrates, the biosynthesis of steroid hormones is initiated by cytochrome P450 CYP11A1 which converts cholesterol to pregnenolone.
[67]
Aminoglutethimide: review of pharmacology and clinical use - PubMed
Aminoglutethimide is indicated for treatment of certain patients with Cushing's syndrome and breast cancer. Other potential uses (prostate carcinoma, low renin ...
[68]
Medical adrenalectomy in patients with advanced breast cancer
Medical adrenalectomy, consisting of aminoglutethimide plus either dexamethasone or hydrocortisone, was administered to 53 women with advanced breast cancer.
[69]
Treatment of Cushing's disease: a mechanistic update in
Ketoconazole inhibits the side-chain cleavage complex (P450scc, CYP11A1, or 20,22 desmolase), 11β-hydroxylase, and 17α-hydroxylase (Table 4) (Feldman 1986 ...
[70]
Tetrandrine inhibits aldosterone synthesis by covalently targeting ...
Jun 13, 2024 · This interaction alters the stability of heme within CYP11A1, subsequently impeding electron transfer and inhibiting aldosterone biosynthesis.
[71]
Targeted Inhibition of CYP11A1 in Castration-Resistant Prostate ...
Dec 26, 2023 · Prostate cancer is regulated by steroid hormones, even in castration-resistant disease. ODM-208, a novel inhibitor of cytochrome P450 11A1 ...