Binding of steroid substrates reveals the key to the productive transition of the cytochrome P450 OleP.

IF 4.4 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Structure Pub Date : 2024-09-05 Epub Date: 2024-07-05 DOI:10.1016/j.str.2024.06.005
Antonella Costanzo, Francesca Fata, Ida Freda, Maria Laura De Sciscio, Elena Gugole, Giovanni Bulfaro, Matteo Di Renzo, Luca Barbizzi, Cécile Exertier, Giacomo Parisi, Marco D'Abramo, Beatrice Vallone, Carmelinda Savino, Linda Celeste Montemiglio
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Abstract

OleP is a bacterial cytochrome P450 involved in oleandomycin biosynthesis as it catalyzes regioselective epoxidation on macrolide intermediates. OleP has recently been reported to convert lithocholic acid (LCA) into murideoxycholic acid through a highly regioselective reaction and to unspecifically hydroxylate testosterone (TES). Since LCA and TES mainly differ by the substituent group at the C17, here we used X-ray crystallography, equilibrium binding assays, and molecular dynamics simulations to investigate the molecular basis of the diverse reactivity observed with the two steroids. We found that the differences in the structure of TES and LCA affect the capability of these molecules to directly form hydrogen bonds with N-terminal residues of OleP internal helix I. The establishment of these contacts, by promoting the bending of helix I, fosters an efficient trigger of the open-to-closed structural transition that occurs upon substrate binding to OleP and contributes to the selectivity of the subsequent monooxygenation reaction.

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类固醇底物的结合揭示了细胞色素 P450 OleP 生产转换的关键。
OleP 是一种细菌细胞色素 P450,参与油霉素的生物合成,催化大环内酯中间体的区域选择性环氧化反应。最近有报道称,OleP 可通过高区域选择性反应将石胆酸(LCA)转化为鼠去氧胆酸,并对睾酮(TES)进行非特异性羟化。由于 LCA 和 TES 的主要区别在于 C17 上的取代基,因此我们在此使用 X 射线晶体学、平衡结合试验和分子动力学模拟来研究观察到的这两种类固醇的不同反应性的分子基础。我们发现,TES 和 LCA 结构的差异影响了这些分子与 OleP 内部螺旋 I 的 N 端残基直接形成氢键的能力。这些接触的建立促进了螺旋 I 的弯曲,从而有效地触发了底物与 OleP 结合后发生的从开放到封闭的结构转变,并提高了后续单氧反应的选择性。
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来源期刊
Structure
Structure 生物-生化与分子生物学
CiteScore
8.90
自引率
1.80%
发文量
155
审稿时长
3-8 weeks
期刊介绍: Structure aims to publish papers of exceptional interest in the field of structural biology. The journal strives to be essential reading for structural biologists, as well as biologists and biochemists that are interested in macromolecular structure and function. Structure strongly encourages the submission of manuscripts that present structural and molecular insights into biological function and mechanism. Other reports that address fundamental questions in structural biology, such as structure-based examinations of protein evolution, folding, and/or design, will also be considered. We will consider the application of any method, experimental or computational, at high or low resolution, to conduct structural investigations, as long as the method is appropriate for the biological, functional, and mechanistic question(s) being addressed. Likewise, reports describing single-molecule analysis of biological mechanisms are welcome. In general, the editors encourage submission of experimental structural studies that are enriched by an analysis of structure-activity relationships and will not consider studies that solely report structural information unless the structure or analysis is of exceptional and broad interest. Studies reporting only homology models, de novo models, or molecular dynamics simulations are also discouraged unless the models are informed by or validated by novel experimental data; rationalization of a large body of existing experimental evidence and making testable predictions based on a model or simulation is often not considered sufficient.
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