首个与细胞色素P450形成的氧化还原复合物揭示了3Fe-4S铁氧化还原蛋白多样性的结构见解。

Structural insights into 3Fe-4S ferredoxins diversity in highlighted by a first redox complex with P450.

作者信息

Gilep Andrei, Varaksa Tatsiana, Bukhdruker Sergey, Kavaleuski Anton, Ryzhykau Yury, Smolskaya Sviatlana, Sushko Tatsiana, Tsumoto Kouhei, Grabovec Irina, Kapranov Ivan, Okhrimenko Ivan, Marin Egor, Shevtsov Mikhail, Mishin Alexey, Kovalev Kirill, Kuklin Alexander, Gordeliy Valentin, Kaluzhskiy Leonid, Gnedenko Oksana, Yablokov Evgeniy, Ivanov Alexis, Borshchevskiy Valentin, Strushkevich Natallia

机构信息

Institute of Bioorganic Chemistry, National Academy of Sciences of Belarus, Minsk, Belarus.

Laboratory of Intermolecular Interactions, Institute of Biomedical Chemistry, Moscow, Russia.

出版信息

Front Mol Biosci. 2023 Jan 9;9:1100032. doi: 10.3389/fmolb.2022.1100032. eCollection 2022.

DOI:10.3389/fmolb.2022.1100032

PMID:36699703

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9868604/

Abstract

Ferredoxins are small iron-sulfur proteins and key players in essential metabolic pathways. Among all types, 3Fe-4S ferredoxins are less studied mostly due to anaerobic requirements. Their complexes with cytochrome P450 redox partners have not been structurally characterized. In the present work, we solved the structures of both 3Fe-4S ferredoxins from -Fdx alone and the fusion FdxE-CYP143. Our SPR analysis demonstrated a high-affinity binding of FdxE to CYP143. According to SAXS data, the same complex is present in solution. The structure reveals extended multipoint interactions and the shape/charge complementarity of redox partners. Furthermore, FdxE binding induced conformational changes in CYP143 as evident from the solved CYP143 structure alone. The comparison of FdxE-CYP143 and modeled Fdx-CYP51 complexes further revealed the specificity of ferredoxins. Our results illuminate the diversity of electron transfer complexes for the production of different secondary metabolites.

摘要

铁氧化还原蛋白是一类小型铁硫蛋白，也是重要代谢途径中的关键参与者。在所有类型中，3Fe-4S铁氧化还原蛋白由于需要厌氧环境，研究较少。它们与细胞色素P450氧化还原伴侣形成的复合物尚未进行结构表征。在本研究中，我们解析了单独的-Fdx和融合蛋白FdxE-CYP143中3Fe-4S铁氧化还原蛋白的结构。我们的表面等离子体共振（SPR）分析表明FdxE与CYP143具有高亲和力结合。根据小角X射线散射（SAXS）数据，溶液中存在相同的复合物。该结构揭示了氧化还原伴侣之间广泛的多点相互作用以及形状/电荷互补性。此外，仅从解析的CYP143结构就可以明显看出，FdxE的结合诱导了CYP143的构象变化。FdxE-CYP143与模拟的Fdx-CYP51复合物的比较进一步揭示了铁氧化还原蛋白的特异性。我们的结果阐明了用于产生不同次生代谢物的电子传递复合物的多样性。