Identification of a Potent and Broad-Spectrum Hepatitis C Virus Fusion Inhibitory Peptide from the E2 Stem Domain.
Identification of a Potent and Broad-Spectrum Hepatitis C Virus Fusion Inhibitory Peptide from the E2 Stem Domain.
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从 E2 干结构域中鉴定出有效的广谱丙型肝炎病毒融合抑制肽
DOI：
10.1038/srep25224
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发表时间：
2016-04-28
影响因子：
4.6
通讯作者：
Yang W
中科院分区：
文献类型：
--
作者：
Chi X;Niu Y;Cheng M;Liu X;Feng Y;Zheng F;Fan J;Li X;Jin Q;Zhong J;Li YP;Yang W
丙型肝炎病毒(HCV)包膜蛋白E1和E2在病毒进入过程中起着至关重要的作用。然而，丙型肝炎病毒的融合机制在很大程度上仍不清楚，这阻碍了有效融合抑制剂的开发。在这里，我们开发了两种基于细胞的膜融合模型，允许筛选覆盖全长E1和E2氨基酸序列的肽库。研究发现，E27具有阻断E1E2介导的细胞-细胞融合、抑制丙型肝炎病毒假颗粒进入细胞和抑制细胞培养来源的丙型肝炎病毒感染的能力。E27对主要的基因1-6有广谱抑制作用。添加时间实验表明，E27主要在丙型肝炎病毒进入的后期发挥作用，而不影响丙型肝炎病毒共受体的表达和定位。此外，我们还证实了E27干扰了异位表达的E1E2在细胞中的异源二聚化，突变分析表明E27可能针对E1中的一个保守区。综上所述，我们的发现为开发有效和广谱的丙型肝炎病毒融合抑制剂提供了一个新的候选方案和策略，可以补充目前治疗慢性丙型肝炎的直接作用的抗病毒药物，并揭示丙型肝炎病毒膜融合的机制。
Hepatitis C virus (HCV) envelope proteins E1 and E2 play an essential role in virus entry. However, the fusion mechanisms of HCV remain largely unclear, hampering the development of efficient fusion inhibitors. Here, we developed two cell-based membrane fusion models that allow for screening a peptide library covering the full-length E1 and E2 amino acid sequences. A peptide from the E2 stem domain, named E27, was found to possess the ability to block E1E2-mediated cell-cell fusion and inhibit cell entry of HCV pseudoparticles and infection of cell culture-derived HCV at nanomolar concentrations. E27 demonstrated broad-spectrum inhibition of the major genotypes 1 to 6. A time-of-addition experiment revealed that E27 predominantly functions in the late steps during HCV entry, without influencing the expression and localization of HCV co-receptors. Moreover, we demonstrated that E27 interfered with hetero-dimerization of ectopically expressed E1E2 in cells and mutational analysis suggested that E27 might target a conserved region in E1. Taken together, our findings provide a novel candidate as well as a strategy for developing potent and broad-spectrum HCV fusion inhibitors, which may complement the current direct-acting antiviral medications for chronic hepatitis C and shed light on the mechanism of HCV membrane fusion.