Cyp17a1

CYP17A1
  • 文章类型: Journal Article
    人细胞色素P450(CYP)酶由执行单加氧酶活性的57种单独的酶组成。它们在代谢异源生物和产生重要的内源性化合物方面具有不同的生理作用,如类固醇激素和维生素。至少七种CYP酶参与类固醇生物合成。类固醇生成主要发生在肾上腺和性腺,将每个反应连接到底物和产物。类固醇对于维持生命至关重要,并显着有助于体内的性分化和生殖功能。类固醇生物合成疾病经常会导致严重的健康问题,并导致疾病的发展。比如前列腺癌,乳腺癌,和库欣综合征。在这次审查中,我们提供有关类固醇生物合成过程中涉及的主要CYP酶的最新知识,关于它们的酶促机制和对开发新候选药物的临床意义。
    Human cytochrome P450 (CYP) enzymes are composed of 57 individual enzymes that perform monooxygenase activities. They have diverse physiological roles in metabolizing xenobiotics and producing important endogenous compounds, such as steroid hormones and vitamins. At least seven CYP enzymes are involved in steroid biosynthesis. Steroidogenesis primarily occurs in the adrenal glands and gonads, connecting each reaction to substrates and products. Steroids are essential for maintaining life and significantly contribute to sexual differentiation and reproductive functions within the body. Disorders in steroid biosynthesis can frequently cause serious health problems and lead to the development of diseases, such as prostate cancer, breast cancer, and Cushing\'s syndrome. In this review, we provide current updated knowledge on the major CYP enzymes involved in the biosynthetic process of steroids, with respect to their enzymatic mechanisms and clinical implications for the development of new drug candidates.
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  • 文章类型: Journal Article
    Testosterone is closely associated with lipid metabolism and known to affect body fat composition and muscle mass in males. However, the mechanisms by which testosterone acts on lipid metabolism are not yet fully understood, especially in teleosts. In this study, cyp17a1-/- zebrafish ( Danio rerio) exhibited excessive visceral adipose tissue (VAT), lipid content, and up-regulated expression and activity of hepatic de novo lipogenesis (DNL) enzymes. The assay for transposase accessible chromatin with sequencing (ATAC-seq) results demonstrated that chromatin accessibility of DNL genes was increased in cyp17a1-/- fish compared to cyp17a1+/+ male fish, including stearoyl-CoA desaturase ( scd) and fatty acid synthase ( fasn). Androgen response element (ARE) motifs in the androgen signaling pathway were significantly enriched in cyp17a1+/+ male fish but not in cyp17a1-/- fish. Both androgen receptor ( ar)-/- and wild-type (WT) zebrafish administered with Ar antagonist flutamide displayed excessive visceral adipose tissue, lipid content, and up-regulated expression and activity of hepatic de novo lipogenesis enzymes. The Ar agonist BMS-564929 reduced the content of VAT and lipid content, and down-regulated acetyl-CoA carboxylase a ( acaca), fasn, and scd expression. Mechanistically, the rescue effect of testosterone on cyp17a1-/- fish in terms of phenotypes was abolished when ar was additionally depleted. Collectively, these findings reveal that testosterone inhibits lipid deposition by down-regulating DNL genes via Ar in zebrafish, thus expanding our understanding of the relationship between testosterone and lipid metabolism in teleosts.
    睾酮可以影响雄性体内脂肪成分和肌肉质量,与脂质代谢密切相关。然而,在硬骨鱼中,睾酮对脂质代谢的作用机制尚不清晰。在该研究中,我们首先观察到雄激素合成的 cyp17a1-/-斑马鱼表现出增多的内脏脂肪组织(VAT),其脂肪含量以及肝脏脂肪从头合成(DNL)酶的表达和活性上调。染色质转座酶可及性测序分析(ATAC-seq)的结果表明,与 cyp17a1+/+雄鱼相比, cyp17a1-/-鱼的DNL基因的染色质可及性增加,包括硬脂酰辅酶a去饱和酶( scd)和脂肪酸合成酶( fasn)。雄激素信号通路中的雄激素反应元件(ARE)基序在 cyp17a1+/+雄鱼中显著富集,但未在 cyp17a1-/-鱼中富集。同样地,雄激素受体( ar)-/-斑马鱼和雄激素受体拮抗剂氟他胺(Flutamide)处理的野生型斑马鱼也表现出VAT增多和脂质含量增加,肝脏脂肪从头合成酶的表达和活性上调。相反,雄激素受体激动剂BMS-564929显著减少了VAT和脂质含量,下调了乙酰辅酶a羧化酶a( acaca)、 fasn和 scd的表达。有趣的是,睾酮处理可以有效挽救 cyp17a1-/-斑马鱼的上述表型,但在 ar被额外敲除后(即在 cyp17a1-/-; ar-/-斑马鱼中)则未见挽救效果。综上所述,我们的研究揭示了睾酮通过Ar下调DNL基因表达和活性,进而抑制斑马鱼的脂质沉积。该研究有助于深入理解硬骨鱼类的雄激素调节脂质代谢的分子机制。.
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  • 文章类型: Journal Article
    内分泌干扰化学物质(EDC)可能通过改变类固醇代谢来影响前列腺癌(PCa)的发展。尽管它们控制肿瘤生长的确切作用机制尚不清楚,EDC可抑制类固醇生成酶,如CYP17A1或CYP19A1,其参与雄激素或雌激素的产生。高水平的循环雄激素与男性的PCa和女性的多囊卵巢综合征(PCOS)有关。精油或其代谢物,像薰衣草油和茶树油,据报道,由于暴露于薰衣草类香料,男孩青春期前男性乳房发育症和女孩早产性类固醇失衡。我们筛选了一系列EO成分以确定它们对CYP17A1和CYP19A1的影响。进行计算对接以预测精油与CYP17A1和CYP19A1的结合。使用放射性标记的底物或液相色谱-高分辨率质谱进行功能测定,并在LNCaP细胞中进行细胞活力测定。许多测试化合物结合接近CYP17A1的活性位点,并且(+)-Cedrol具有与CYP17A1和CYP19A1的最佳结合。桉树脑,二氢-β-离子酮,和(-)-α-pine烯显示20%至40%的脱氢表雄酮产生抑制;一些化合物也影响CYP19A1。在各种美容和卫生用品中广泛使用这些精油是常见的,但对其潜在有害副作用的了解有限。我们的结果表明,长时间接触这些精油可能会导致类固醇失衡。另一方面,由于它们对降低雄激素输出和在类固醇细胞色素P450的活性位点结合的能力的作用,这些化合物可能为抗PCa和PCOS等高雄激素性疾病的新型化合物提供设计思路。
    Endocrine-disrupting chemicals (EDCs) may impact the development of prostate cancer (PCa) by altering the steroid metabolism. Although their exact mechanism of action in controlling tumor growth is not known, EDCs may inhibit steroidogenic enzymes such as CYP17A1 or CYP19A1 which are involved in the production of androgens or estrogens. High levels of circulating androgens are linked to PCa in men and Polycystic Ovary Syndrome (PCOS) in women. Essential oils or their metabolites, like lavender oil and tea tree oil, have been reported to act as potential EDCs and contribute towards sex steroid imbalance in cases of prepubertal gynecomastia in boys and premature thelarche in girls due to the exposure to lavender-based fragrances. We screened a range of EO components to determine their effects on CYP17A1 and CYP19A1. Computational docking was performed to predict the binding of essential oils with CYP17A1 and CYP19A1. Functional assays were performed using the radiolabeled substrates or Liquid Chromatography-High-Resolution Mass Spectrometry and cell viability assays were carried out in LNCaP cells. Many of the tested compounds bind close to the active site of CYP17A1, and (+)-Cedrol had the best binding with CYP17A1 and CYP19A1. Eucalyptol, Dihydro-β-Ionone, and (-)-α-pinene showed 20% to 40% inhibition of dehydroepiandrosterone production; and some compounds also effected CYP19A1. Extensive use of these essential oils in various beauty and hygiene products is common, but only limited knowledge about their potential detrimental side effects exists. Our results suggest that prolonged exposure to some of these essential oils may result in steroid imbalances. On the other hand, due to their effect on lowering androgen output and ability to bind at the active site of steroidogenic cytochrome P450s, these compounds may provide design ideas for novel compounds against hyperandrogenic disorders such as PCa and PCOS.
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  • 文章类型: Journal Article
    先前的体内和体外研究表明,LH/hCG治疗后,大鼠排卵前颗粒细胞(GC)中Krüppel样因子4(Klf4)的显着上调,及其在调节黄体移位期间Cyp19A1表达中的作用在类固醇生成中。在这项研究中,我们检测了Klf4是否也介导了LH诱导的原代大鼠排卵前GCs中Cyp17A1表达抑制。在体外对GC的LH治疗的反应中,Cyp17A1表达下降到不到其初始值的一半,1小时,培养24小时保持低位。Klf4的过表达降低了基础和Sf1诱导的Cyp17A1表达并增加了孕酮分泌。通过siRNA减少内源性Klf4升高基础Cyp17A1表达,但不影响LH刺激的孕酮产生。Klf4的过表达也显著减弱了Sf1诱导的Cyp17A1启动子活性。另一方面,启动子中保守的Sp1/Klf结合基序的突变表明,该基序不是Klf4介导的抑制所必需的。一起来看,这些数据表明Cyp17A1基因可能是排卵前GCs中LH诱导的Klf4的下游靶标之一。这些信息可能有助于确定预防高雄激素性疾病中发生的分子变化的潜在靶标。
    Previous in vivo and in vitro studies have demonstrated a dramatic up-regulation of Krüppel-like factor 4 (Klf4) in rat preovulatory granulosa cells (GCs) after LH/hCG treatment and its role in regulating Cyp19A1 expression during the luteal shift in steroidogenesis. In this study, we examined whether Klf4 also mediates the LH-induced repression of Cyp17A1 expression in primary rat preovulatory GCs. In response to LH treatment of GCs in vitro, Cyp17A1 expression declined to less than half of its initial value by 1 h, remaining low for 24 h of culture. Overexpression of Klf4 decreased basal and Sf1-induced Cyp17A1 expressions and increased progesterone secretion. Reduction of endogenous Klf4 by siRNA elevated basal Cyp17A1 expression but did not affect LH-stimulated progesterone production. Overexpression of Klf4 also significantly attenuated Sf1-induced Cyp17A1 promoter activity. On the other hand, mutation of the conserved Sp1/Klf binding motif in the promoter revealed that this motif is not required for Klf4-mediated repression. Taken together, these data indicate that the Cyp17A1 gene may be one of the downstream targets of Klf4, which is induced by LH in preovulatory GCs. This information may help in identifying potential targets for preventing the molecular changes occurring in hyperandrogenic disorders.
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  • 文章类型: Journal Article
    多囊卵巢综合征(PCOS)的定义特征包括类固醇生成基因的表达升高,卵泡膜细胞雄激素生物合成和雄激素的外周水平。在先前的研究中,我们将血管细胞粘附分子1(VCAM1)鉴定为特定NR2F2/SF1(/)卵泡膜细胞中的选择性雄激素靶基因。通过在小鼠CYP17A1卵泡膜细胞中选择性删除NR2F2和VCAM1,我们记录了NR2F2和VCAM1影响不同的有时相反的卵泡膜细胞功能,改变体内卵泡发育:包括卵巢形态的主要变化,类固醇生成,基因表达谱,免疫定位图像(NR5A1,CYP11A1,NOTCH1,CYP17A1,INSL3,VCAM1,NR2F2)以及颗粒细胞功能。我们建议卵泡膜细胞通过调节雄激素的产生和作用来影响卵泡的完整性,以及颗粒细胞分化/黄体化反应的雄激素和促性腺激素可能是PCOS的基础。
    Defining features of polycystic ovary syndrome (PCOS) include elevated expression of steroidogenic genes, theca cell androgen biosynthesis, and peripheral levels of androgens. In previous studies, we identified vascular cell adhesion molecule 1 (VCAM1) as a selective androgen target gene in specific NR2F2/SF1 (+/+) theca cells. By deleting NR2F2 and VCAM1 selectively in CYP17A1 theca cells in mice, we documented that NR2F2 and VCAM1 impact distinct and sometimes opposing theca cell functions that alter ovarian follicular development in vivo: including major changes in ovarian morphology, steroidogenesis, gene expression profiles, immunolocalization images (NR5A1, CYP11A1, NOTCH1, CYP17A1, INSL3, VCAM1, NR2F2) as well as granulosa cell functions. We propose that theca cells impact follicle integrity by regulating androgen production and action, as well as granulosa cell differentiation/luteinization in response to androgens and gonadotropins that may underlie PCOS.
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  • 文章类型: Journal Article
    六种细胞色素P450酶参与人类类固醇生成,将胆固醇转化为性类固醇,盐皮质激素,和糖皮质激素。虽然早期的工作是用更容易获得的来源的类固醇生成P450直系同源物完成的,通过成功的药物设计的基础生物化学知识已经大大促进了重组表达,这些膜蛋白的高度纯化的人类版本。许多膜蛋白难以表达和纯化并且不稳定。通过修饰和/或截短与膜相互作用的N-末端,促进了P450在大肠杆菌中的表达。而金属亲和树脂和组氨酸标记大大促进了纯化。然而,大量的优化仍然经常需要维持蛋白质的稳定性。随着时间的推移,已开发并调整了通用的三柱纯化方案,以产生大量的完全活性,高度纯化的人类细胞色素P450酶使许多结构的应用成为可能,生物化学,和生物物理技术来阐明这些关键人类酶的奥秘。
    Six cytochrome P450 enzymes are involved in human steroidogenesis, converting cholesterol to sex steroids, mineralocorticoids, and glucocorticoids. While early work was accomplished with steroidogenic P450 orthologs from more accessible sources, knowledge of basic biochemistry through successful drug design have been greatly facilitated by recombinantly-expressed, highly purified human versions of these membrane proteins. Many membrane proteins are difficult to express and purify and are unstable. Membrane P450 expression in E. coli has been facilitated by modification and/or truncation of the membrane-interacting N-terminus, while metal-affinity resins and histidine-tagging greatly facilitates purification. However, substantial optimization is still frequently required to maintain protein stability. Over time, a generalized three-column purification scheme has been developed and tweaked to generate substantial quantities of fully active, highly purified human cytochrome P450 enzymes that have made possible the application of many structural, biochemical, and biophysical techniques to elucidate the mysteries of these critical human enzymes.
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  • 文章类型: Journal Article
    这项研究报道了用带有硫基取代基的苯基取代细胞色素P45017α-羟化酶/12,20-裂解酶(CYP17A1)抑制剂化学主链结构上含氮杂环的新型化合物的合成和评估。初步筛选显示化合物对CYP17A1活性具有显著抑制。此后确定化合物对细胞色素P45021-羟化酶的选择性,细胞色素P4503A4和细胞色素P450氧化还原酶。此外,这些化合物对aldo-keto还原酶1C3(AKR1C3)显示出弱的抑制活性。还评估了化合物对类固醇激素水平的影响,观察到一些显著的调节作用。这项工作为开发更有效的特异性靶向CYP17A1和AKR1C3的双重抑制剂铺平了道路。
    This study reports on the synthesis and evaluation of novel compounds replacing the nitrogen-containing heterocyclic ring on the chemical backbone structure of cytochrome P450 17α-hydroxylase/12,20-lyase (CYP17A1) inhibitors with a phenyl bearing a sulfur-based substituent. Initial screening revealed compounds with marked inhibition of CYP17A1 activity. The selectivity of compounds was thereafter determined against cytochrome P450 21-hydroxylase, cytochrome P450 3A4, and cytochrome P450 oxidoreductase. Additionally, the compounds showed weak inhibitory activity against aldo-keto reductase 1C3 (AKR1C3). The compounds\' impact on steroid hormone levels was also assessed, with some notable modulatory effects observed. This work paves the way for developing more potent dual inhibitors specifically targeting CYP17A1 and AKR1C3.
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  • 文章类型: Journal Article
    背景:冠状动脉疾病(CAD)是一种多因素复杂的特征,可遗传,尤其是在早发家庭。然而,影响早发性CAD易感性的遗传因素尚未完全表征.
    方法:在本研究中,我们在一个中国汉族CAD家族的CYP17A1基因中发现了一个罕见的无义变异。为了验证这种变异对动脉粥样硬化和早发性冠状动脉疾病的影响,我们对人口进行了研究,细胞,和老鼠。
    结果:该突变在所有受影响的家庭成员中与临床综合征精确聚集,在未受影响的家庭成员和无关的对照中不存在。类似于人类表型,CYP17A1缺陷小鼠呈现代谢综合征伴高血压的表型,血清葡萄糖浓度升高,以及中心性肥胖和脂肪肝的表现。此外,CYP17A1敲除小鼠或CYP17A1+ApoE双敲除小鼠比野生型(WT)高脂日记发展更多的动脉粥样硬化病变。在细胞模型中,发现CYP17A1通过增加葡萄糖的摄入和利用而参与葡萄糖代谢。通过激活IGF1/mTOR/HIF1-α信号的方式,这与CYP17A1敲除小鼠糖耐量受损和胰岛素抵抗一致。
    结论:通过我们对细胞的研究,老鼠和人类,我们将CYP17A1鉴定为参与动脉粥样硬化过程病理生理学的关键蛋白,并揭示了CYP17A1C987X突变诱导动脉粥样硬化和涉及葡萄糖稳态调节的早发性CAD的可能机制.视频摘要。
    Coronary artery disease (CAD) is a multi-factor complex trait and is heritable, especially in early-onset families. However, the genetic factors affecting the susceptibility of early-onset CAD are not fully characterized.
    In the present study, we identified a rare nonsense variant in the CYP17A1 gene from a Chinese Han family with CAD. To validate the effect of this variation on atherosclerosis and early-onset coronary artery disease, we conducted studies on population, cells, and mice.
    The mutation precisely congregated with the clinical syndrome in all the affected family members and was absent in unaffected family members and unrelated controls. Similar to the human phenotype, the CYP17A1-deficient mice present the phenotype of metabolic syndrome with hypertension, increased serum glucose concentration, and presentation of central obesity and fatty liver. Furthermore, CYP17A1 knockout mice or CYP17A1 + ApoE double knockout mice developed more atherosclerotic lesions than wild type (WT) with high fat diary. In cell models, CYP17A1 was found to be involved in glucose metabolism by increasing glucose intake and utilization, through activating IGF1/mTOR/HIF1-α signaling way, which was consistent in CYP17A1 knockout mice with impaired glucose tolerance and insulin resistance.
    Through our study of cells, mice and humans, we identified CYP17A1 as a key protein participating in the pathophysiology of the atherosclerotic process and the possible mechanism of CYP17A1 C987X mutation induced atherosclerosis and early-onset CAD involving glucose homeostasis regulation was revealed. Video Abstract.
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  • 文章类型: Journal Article
    细胞色素P450(CYP)的催化循环需要来自蛋白质氧化还原伴侣的两个电子和来自水的两个质子来产生主要的催化中间体,血红素卟啉环上有π阳离子的铁-氧代络合物,称为化合物1。质子化步骤至少是部分限速的,因此,P450催化的稳态速率在氘代溶剂(D2O)中通常慢1.5-3倍。然而,在几个P450系统中,观察到明显的逆动力学溶剂同位素效应(KSIE~0.4-0.7),在D2O中反应更快。这提出了一个重要的机制问题:这种反溶剂同位素效应是否与化合物1催化反应相容,还是表明涉及另一种催化中间体?在本通讯中,我们使用P450稳态动力学的详尽数值模型来证明,对于纯化合物1驱动的P450催化循环,无法获得显着的逆KSIE。相反,另一种选择,质子化独立,需要引入催化中间体。这一结果适用于自然界中广谱的P450,但作为一个例子,我们在人类类固醇生物合成P450CYP17A1中使用了广泛记录的反同位素效应,其中已表征了血红素过氧阴离子中间体的参与。基于这一分析,我们表明,反向KSIE的观察可以用作细胞色素P450中反应循环中间体的一般机理探针。
    The catalytic cycle of the cytochromes P450 (CYP) requires two electrons from a protein redox partner and two protons from water to generate the main catalytic intermediate, a ferryl-oxo complex with π-cation on the heme porphyrin ring, termed Compound 1. The protonation steps are at least partially rate-limiting, therefore the steady-state rates of P450 catalysis are usually slower in deuterated solvent (D2O) by a factor of 1.5-3. However, in several P450 systems a pronounced inverse kinetic solvent isotope effect (KSIE ∼0.4-0.7) is observed, where the reaction is faster in D2O. This raises an important mechanistic question: Is this inverse solvent isotope effect compatible with Compound 1 catalyzed reactions, or is it indicative of another catalytic intermediate being involved? In this communication we use exhaustive numerical modeling of the P450 steady-state kinetics to demonstrate that a significant inverse KSIE cannot be obtained for a pure Compound 1 driven catalytic cycle of P450. Rather, an alternative, protonation independent, catalytic intermediate needs to be introduced. This result is applicable to the broad spectrum of P450s in nature, but as an example we use the extensively documented inverse isotope effect in the human steroid biosynthetic P450 CYP17A1 where the involvement of a heme peroxo anion intermediate has been characterized. Based on this analysis, we show that the observation of an inverse KSIE can be used as a general mechanistic probe for reaction cycle intermediates in the cytochromes P450.
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  • 文章类型: Journal Article
    寻找盐皮质激素以解释某些醛固酮水平受抑制的低肾素高血压病例,导致人尿液中丰富的类固醇18-羟基皮质醇的分离。18-羟基皮质醇被证明是不活跃的,但是由于它与醛固酮合成的前体相似,牛蛙的肾上腺与皮质醇一起孵育,结果发现了结构上与醛固酮相似的18-氧代皮质醇,但是有一个像皮质醇一样的17α-羟基。18-氧化皮质醇是一种弱的盐皮质激素。它的合成主要发生在肾小球带中,其中CYP11B2(醛固酮合酶)和CYP17A1(17α-羟化酶)的共表达发生在可变数量的细胞中。测量18-氧皮质醇的临床价值是它可以区分原发性醛固酮增多症的亚型。与特发性双侧醛固酮增多症患者相比,醛固酮分泌腺瘤患者的体细胞突变类型显着升高,并有助于预测醛固酮分泌腺瘤的体细胞突变类型。因为与其他基因突变相比,具有KCNJ5突变的基因更高。
    The search for mineralocorticoids to explain some cases of low renin hypertension with suppressed aldosterone levels led to the isolation of the abundant steroid 18-hydroxycortisol in human urine. 18-Hydroxycortisol proved to be inactive, but because of its similarity to precursors for the synthesis of aldosterone, bullfrog adrenals were incubated with cortisol, resulting in the discovery of 18-oxocortisol which is structurally similar to aldosterone, but with a 17α-hydroxy group like cortisol. 18-Oxocortisol is a weak mineralocorticoid. Its synthesis occurs primarily in the zona glomerulosa where co-expression of the CYP11B2 (aldosterone synthase) and the CYP17A1 (17α-hydroxylase) occurs in a variable number of cells. The clinical value of the measurement of 18-oxocortisol is that it serves to distinguish subtypes of primary aldosteronism. It is significantly elevated in patients with aldosterone-producing adenomas in comparison to those with idiopathic bilateral hyperaldosteronism and helps predict the type of somatic mutation in the aldosterone-producing adenomas, as it is higher in those with KCNJ5 mutations compared to other gene mutations.
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