The steroid metabolite 16(β)-OH-androstenedione generated by CYP21A2 serves as a substrate for CYP19A1
The steroid metabolite 16(β)-OH-androstenedione generated by CYP21A2 serves as a substrate for CYP19A1
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CYP21A2 产生的类固醇代谢物 16(β)-OH-androstenedione 作为 CYP19A1 的底物
DOI：
10.1016/j.jsbmb.2017.01.002
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Bernhardt R
中科院分区：
文献类型：
--
作者：
Neunzig J;Milhim M;Schiffer L;Kathri Y;Zapp J;Sánchez-Guijo A;Hartmann MF;Wudy SA;Bernhardt R
21-羟化酶（CYP 21 A2）是一种类固醇生成酶，对合成矿物质和糖皮质激素至关重要。它被描述为通过在位置C21处的羟基化将孕酮以及17-OH-孕酮分别转化为11-脱氧皮质酮（DOC）和11-脱氧皮质醇（RSS）。在这项研究中，我们揭示了CYP 21 A2具有比假设更广泛的类固醇底物谱。利用由纯化的人CYP 21 A2和人细胞色素P450还原酶（CPR）组成的重组体外系统，我们证明了CYP 21 A2能够代谢DOC、RSS、雄烯二酮（A4）和睾酮（T）。此外，A4的转化产生了一种产物，其结构通过NMR光谱进行了解析，显示在C16-β位发生了羟基化。研究了这种类固醇代谢物16（β）-OH-雄烯二酮（16 bOHA 4）的雄激素性质，并与A4进行了比较。两种类固醇代谢产物均显示为人雄激素受体的弱激动剂。此外，将16 bOHA 4与芳香酶（CYP 19 A1）的相互作用与A4的相互作用进行了比较，表明C16羟基不影响与CYP 19 A1的结合。与此相反，动力学参数的阐明表明，增加的Kmandreducedkcat值导致催化效率降低2倍相比，A4。这些发现与我们的对接研究一致，揭示了两种类固醇与血红素铁的相似结合构象和距离。此外，研究了16 bOHA 4的产物，推测为16-羟基-雌酮（16 bOHE 1）的雌激素活性，与雌二醇和雌酮相比，其雌激素活性可忽略不计。最后，发现16 bOHA 4存在于11-羟化酶缺乏症患者和内分泌肿瘤患者中。总之，这项研究提供了新的信息，类固醇激素的生物合成，并提出了一种新的方法来检测进一步潜在的相关新的类固醇代谢产物。
The 21-hydroxylase (CYP21A2) is a steroidogenic enzyme crucial for the synthesis of mineralo- and glucocorticoids. It is described to convert progesterone as well as 17-OH-progesterone, through a hydroxylation at position C21, into 11-deoxycorticosterone (DOC) and 11-deoxycortisol (RSS), respectively. In this study we unraveled CYP21A2 to have a broader steroid substrate spectrum than assumed. Utilizing a reconstitutedin vitrosystem, consisting of purified human CYP21A2 and human cytochrome P450 reductase (CPR) we demonstrated that CYP21A2 is capable to metabolize DOC, RSS, androstenedione (A4) and testosterone (T). In addition, the conversion of A4 rendered a product whose structure was elucidated through NMR spectroscopy, showing a hydroxylation at position C16-beta. The androgenic properties of this steroid metabolite, 16(β)-OH-androstenedione (16bOHA4), were investigated and compared with A4. Both steroid metabolites were shown to be weak agonists for the human androgen receptor. Moreover, the interaction of 16bOHA4 with the aromatase (CYP19A1) was compared to that of A4, indicating that the C16 hydroxyl group does not influence the binding with CYP19A1. In contrast, the elucidation of the kinetic parameters showed an increasedKmand decreasedkcatvalue resulting in a 2-fold decreased catalytic efficiency compared to A4. These findings were in accordance with our docking studies, revealing a similar binding conformation and distance to the heme iron of both steroids. Furthermore, the product of 16bOHA4, presumably 16-hydroxy-estrone (16bOHE1), was investigated with regard to its estrogenic activity, which was negligible compared to estradiol and estrone. Finally, 16bOHA4 was found to be present in a patient with 11-hydroxylase deficiency and in a patient with an endocrine tumor. Taken together, this study provides novel information on the steroid hormone biosynthesis and presents a new method to detect further potential relevant novel steroid metabolites.
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细胞色素 P450 和类固醇激素生物合成
DOI：
--
发表时间：
2007
期刊：
影响因子：
--
作者：
R. Bernhardt;M. Waterman
通讯作者：
M. Waterman
DOI：
10.1016/j.bbrc.2006.03.067
发表时间：
2006-05-26
影响因子：
3.1
作者：
Arase, M;Waterman, MR;Kagawa, N
通讯作者：
Kagawa, N
DOI：
10.3390/molecules181113228
发表时间：
2013-10-25
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Bloem LM;Storbeck KH;Schloms L;Swart AC
通讯作者：
Swart AC
影响因子：
7.3
作者：
Ghosh, Debashis;Lo, Jessica;Morton, Daniel;Valette, Damien;Xi, Jingle;Griswold, Jennifer;Hubbell, Susan;Egbuta, Chinaza;Jiang, Wenhua;An, Jing;Davies, Huw M. L.
通讯作者：
Davies, Huw M. L.
影响因子：
2.8
作者：
Brixius-Anderko, Simone;Hannemann, Frank;Bernhardt, Rita
通讯作者：
Bernhardt, Rita