Identification of Myricetin as an Ebola Virus VP35–Double-Stranded RNA Interaction Inhibitor through a Novel Fluorescence-Based Assay,Biochemistry

当前位置： X-MOL 学术 › Biochemistry › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Identification of Myricetin as an Ebola Virus VP35–Double-Stranded RNA Interaction Inhibitor through a Novel Fluorescence-Based Assay
Biochemistry ( IF 2.9 ) Pub Date : 2018-10-09 00:00:00 , DOI: 10.1021/acs.biochem.8b00892
Gian Luca Daino ₁ , Aldo Frau ₁ , Cinzia Sanna ₁ , Daniela Rigano ₂ , Simona Distinto ₁ , Veronica Madau ₁ , Francesca Esposito ₁ , Elisa Fanunza ₁ , Giulia Bianco ₁ , Orazio Taglialatela-Scafati ₂ , Luca Zinzula ₃ , Elias Maccioni ₁ , Angela Corona ₁ , Enzo Tramontano _{1,

4}

Affiliation

Ebola virus (EBOV) is a filovirus that causes a severe and rapidly progressing hemorrhagic syndrome; a recent epidemic illustrated the urgent need for novel therapeutic agents because no drugs have been approved for treatment of Ebola virus. A key contribution to the high lethality observed during EBOV outbreaks comes from viral evasion of the host antiviral innate immune response in which viral protein VP35 plays a crucial role, blocking interferon type I production, first by masking the viral double-stranded RNA (dsRNA) and preventing its detection by the pattern recognition receptor RIG-I. Aiming to identify inhibitors of the interaction of VP35 with the viral dsRNA, counteracting the VP35 viral innate immune evasion, we established a new methodology for high-yield recombinant VP35 (rVP35) expression and purification and a novel and robust fluorescence-based rVP35–RNA interaction assay (Z′ factor of 0.69). Taking advantage of such newly established methods, we screened a small library of Sardinian natural extracts, identifying Limonium morisianum as the most potent inhibitor extract. A bioguided fractionation led to the identification of myricetin as the component that can inhibit rVP35–dsRNA interaction with an IC₅₀ value of 2.7 μM. Molecular docking studies showed that myricetin interacts with the highly conserved region of the VP35 RNA binding domain, laying the basis for further structural optimization of potent inhibitors of VP35–dsRNA interaction.

中文翻译：

杨梅素作为埃博拉病毒VP35 –双链RNA相互作用抑制剂的鉴定，通过一种新型的基于荧光的分析方法

埃博拉病毒（EBOV）是一种线状病毒，会导致严重且进展迅速的出血综合征。最近的一次流行病表明，迫切需要新型治疗剂，因为尚未批准任何药物可用于治疗埃博拉病毒。EBOV暴发期间观察到的高致死率的关键因素来自病毒的宿主抗病毒先天免疫反应逃逸，其中病毒蛋白VP35起着至关重要的作用，首先通过掩盖病毒双链RNA（dsRNA）来阻断I型干扰素的产生。并防止其被模式识别受体RIG-I检测到。为了确定VP35与病毒dsRNA相互作用的抑制剂，以抵消VP35病毒先天免疫逃逸，Z '系数为0.69）。利用这种新近建立的方法，我们筛选了一个撒丁岛天然提取物的小文库，将莫里斯草（Limonium morisianum）确定为最有效的抑制剂提取物。生物引导分级分离法鉴定了杨梅素是可以抑制rVP35-dsRNA相互作用的成分，IC ₅₀值为2.7μM 。分子对接研究表明，杨梅素与VP35 RNA结合结构域的高度保守区域相互作用，为进一步优化VP35-dsRNA相互作用的有效抑制剂奠定了基础。

更新日期：2018-10-09

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文本刊介绍/投稿指南