Inhibition of SARS CoV Envelope Protein by Flavonoids and Classical Viroporin Inhibitors.
Inhibition of SARS CoV Envelope Protein by Flavonoids and Classical Viroporin Inhibitors.
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DOI：
10.3389/fmicb.2021.692423
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发表时间：
2021
影响因子：
5.2
通讯作者：
Breitinger HG
中科院分区：
文献类型：
--
作者：
Breitinger U;Ali NKM;Sticht H;Breitinger HG
严重急性呼吸综合征冠状病毒（severe acute respiratory syndrome coronavirus，SARS-CoV）是冠状病毒科β冠状病毒属（Betacoronavirus）的一种有包膜的单链正链RNA病毒。SARS-CoV包膜蛋白E是一个小的（约8.4 kDa）通道形成膜蛋白，其序列在SARS-CoV和SARS-CoV-2之间高度保守。作为病毒孔蛋白，它参与病毒生命周期的各个方面，包括组装、出芽、包膜形成、病毒释放和炎性体活化。在这里，SARS-CoV E蛋白在HEK 293细胞中重组表达，并使用膜片钳电生理学和细胞活力测定研究通道活性和病毒孔蛋白抑制剂的作用。我们引入了一个膜导向信号肽，以确保重组E蛋白转移到质膜。E蛋白表达诱导的跨膜电流被各种抑制剂阻断。在离子还原缓冲体系中，电流是质子依赖性的，并被病毒孔蛋白抑制剂金刚烷胺和金刚烷胺阻断。重组E蛋白在高细胞外Na+和高细胞内K+的经典缓冲体系中的I-V关系不具有pH依赖性。金刚烷胺和金刚乙胺以及5-（N，N-氨基）阿米洛利（HMA）可抑制E蛋白介导的电流。我们总共测试了10种黄酮类化合物，发现不同效力的抑制活性。表没食子儿茶素和槲皮素最有效，IC 50值分别为1.5 ± 0.1和3.7 ± 0.2 nM，与金刚乙胺的效力相似（IC 50 = 1.7 ± 0.6 nM）。使用病毒孔蛋白抑制剂的改良细胞活力测定法独立验证膜片钳结果。这些结果有助于开发抑制病毒活性和增殖的新型抗病毒药物。
Severe acute respiratory syndrome coronavirus (SARS-CoV), an enveloped single-stranded positive-sense RNA virus, is a member of the genus Betacoronavirus, family Coronaviridae. The SARS-CoV envelope protein E is a small (∼8.4 kDa) channel-forming membrane protein whose sequence is highly conserved between SARS-CoV and SARS-CoV-2. As a viroporin, it is involved in various aspects of the virus life cycle including assembly, budding, envelope formation, virus release, and inflammasome activation. Here, SARS-CoV E protein was recombinantly expressed in HEK293 cells and channel activity and the effects of viroporin inhibitors studied using patch-clamp electrophysiology and a cell viability assay. We introduced a membrane-directing signal peptide to ensure transfer of recombinant E protein to the plasma membrane. E protein expression induced transmembrane currents that were blocked by various inhibitors. In an ion-reduced buffer system, currents were proton-dependent and blocked by viroporin inhibitors rimantadine and amantadine. I-V relationships of recombinant E protein were not pH-dependent in a classical buffer system with high extracellular Na+ and high intracellular K+. E-protein mediated currents were inhibited by amantadine and rimantadine, as well as 5-(N,N-hexamethylene)amiloride (HMA). We tested a total of 10 flavonoids, finding inhibitory activity of varying potency. Epigallocatechin and quercetin were most effective, with IC50 values of 1.5 ± 0.1 and 3.7 ± 0.2 nM, respectively, similar to the potency of rimantadine (IC50 = 1.7 ± 0.6 nM). Patch-clamp results were independently verified using a modified cell viability assay for viroporin inhibitors. These results contribute to the development of novel antiviral drugs that suppress virus activity and proliferation.
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影响因子：
2.9
作者：
Intharathep, Pathumwadee;Laohpongspaisan, Chittima;Hannongbua, Supot
通讯作者：
Hannongbua, Supot
影响因子：
3.6
作者：
Jalily, Pouria H.;Eldstrom, Jodene;Fedida, David
通讯作者：
Fedida, David
影响因子：
2.7
作者：
González JM;Gomez-Puertas P;Cavanagh D;Gorbalenya AE;Enjuanes L
通讯作者：
Enjuanes L
DOI：
10.1073/pnas.0804958105
发表时间：
2008-08-05
影响因子：
11.1
作者：
Jing, Xianghong;Ma, Chunlong;Lamb, Robert A.
通讯作者：
Lamb, Robert A.
影响因子：
5.6
作者：
Arbely E;Khattari Z;Brotons G;Akkawi M;Salditt T;Arkin IT
通讯作者：
Arkin IT