Fusion core structure of the severe acute respiratory syndrome coronavirus (SARS-CoV): In search of potent SARS-CoV entry inhibitors
Fusion core structure of the severe acute respiratory syndrome coronavirus (SARS-CoV): In search of potent SARS-CoV entry inhibitors
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DOI：
10.1002/jcb.21790
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发表时间：
2008-08-15
影响因子：
4
通讯作者：
Ngai, Sai-Ming
中科院分区：
文献类型：
--
作者：
Chu, Ling-Hon Matthew;Chan, Siu-Hong;Ngai, Sai-Ming
严重急性呼吸道冠状病毒 (SARS-CoV) 刺突 (S) 糖蛋白融合核心由六螺旋束组成，其中三个 C 端七肽重复序列 (HR2) 螺旋与其他三个 N 端七肽重复序列 (HR1) 螺旋的中央卷曲螺旋紧密相连。三个外周 HR2 螺旋中的每一个都与中央 HR1 卷曲螺旋的疏水表面有显着的接触。 HR 螺旋之间协调一致的蛋白质-蛋白质相互作用负责融合事件，从而导致 SARS-CoV 核衣壳释放到目标宿主细胞中。在这项研究中，我们应用重组蛋白和基于合成肽的生物物理测定来表征 HR 螺旋的生物活性。在一项平行实验中，我们采用了 HIV-luc/SARS 假型病毒进入抑制试验来筛选对源自 SARS-CoV S 蛋白 HR 区域的 HR 肽和一系列其他小分子药物的有效抑制活性。三种 HR 肽和五种小分子药物被确定为潜在的抑制剂。 ADS-J1 已被用于干扰 HIV-1 在 CD4(+) 细胞上的融合，在其他小分子药物中表现出最高的 HIV-luc/SARS 假型病毒进入抑制活性。分子模型分析表明，ADS-J1可能与SARS-CoV S HR蛋白中央卷曲螺旋表面疏水沟的深袋结合，阻止SARS-CoV进入宿主细胞。
Severe acute respiratory coronavirus (SARS-CoV) spike (S) glycoprotein fusion core consists of a six-helix bundle with the three C-terminal heptad repeat (HR2) helices packed against a central coiled-coil of the other three N-terminal heptad repeat (HR1) helices. Each of the three peripheral HR2 helices shows prominent contacts with the hydrophobic surface of the central HR1 coiled-coil. The concerted protein-protein interactions among the HR helices are responsible for the fusion event that leads to the release of the SARS-CoV nucleocapsid into the target host-cell. In this investigation, we applied recombinant protein and synthetic peptide-based biophysical assays to characterize the biological activities of the HR helices. In a parallel experiment, we employed a HIV-luc/SARS pseudotyped virus entry inhibition assay to screen for potent inhibitory activities on HR peptides derived from the SARS-CoV S protein HR regions and a series of other small-molecule drugs. Three HR peptides and five small-molecule drugs were identified as potential inhibitors. ADS-J1, which has been used to interfere with the fusogenesis of HIV-1 onto CD4(+) cells, demonstrated the highest HIV-luc/SARS pseudotyped virus-entry inhibition activity among the other small-molecule drugs. Molecular modeling analysis suggested that ADS-J1 may bind to the deep pocket of the hydrophobic groove on the surface of the central coiled-coil of SARS-CoV S HR protein and prevent the entrance of the SARS-CoV into the host cells.