Severe acute respiratory syndrome coroavirus (SARS-CoV) infection inhibition using spike protein heptad repeat-derived peptides
Severe acute respiratory syndrome coroavirus (SARS-CoV) infection inhibition using spike protein heptad repeat-derived peptides
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DOI：
10.1073/pnas.0400576101
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发表时间：
2004-06-01
影响因子：
11.1
通讯作者：
Rottier, PJM
中科院分区：
文献类型：
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作者：
Bosch, BJ;Martina, BEE;Rottier, PJM
冠状病毒SARS- cov是危及生命的严重急性呼吸系统综合征（SARS）的主要病因。为了开发治疗药物，我们测试了从刺突蛋白的近端膜（HR2）和远端膜（HR1）七肽重复区提取的肽作为SARS-CoV感染Vero细胞的抑制剂。HR2肽具有抑制作用，而HR1肽没有。但与小鼠冠状病毒小鼠肝炎病毒（MHV）对应的HR2肽相比，其抑制MHV感染的作用明显降低。生化和电镜分析表明，当混合时，SARS-CoV HR1和HR2肽组装成一个六螺旋束，由HR1作为中心三股螺旋线圈与三个反平行定向的HR2 a-螺旋相结合组成。该复合物的稳定性，通过其耐热解离性来衡量，似乎比相应的MHV复合物低得多，这可能解释了HR2肽的不同抑制能力。与其他I类病毒融合蛋白类似，六螺旋复合体被认为代表了融合后构象，该构象是在将融合肽插入目标膜后形成的，本文提出了位于HR1上游的冠状病毒。融合肽和穗跨膜结构域的紧密结合促进了膜融合。SARS冠状病毒hr2肽的抑制效力为SARS治疗药物的开发提供了有吸引力的基础。
The coronavirus SARS-CoV is the primary cause of the life-threatening severe acute respiratory syndrome (SARS). With the aim of developing therapeutic agents, we have tested peptides derived from the membrane-proximal (HR2) and membrane-distal (HR1) heptad repeat region of the spike protein as inhibitors of SARS-CoV infection of Vero cells. It appeared that HR2 peptides, but not HR1 peptides, were inhibitory. Their efficacy was, however, significantly lower than that of corresponding HR2 peptides of the murine coronavirus mouse hepatitis virus (MHV) in inhibiting MHV infection. Biochemical and electron microscopical analyses showed that, when mixed, SARS-CoV HR1 and HR2 peptides assemble into a six-helix bundle consisting of HR1 as a central triple-stranded coiled coil in association with three HR2 a-helices oriented in an antiparallel manner. The stability of this complex, as measured by its resistance to heat dissociation, appeared to be much lower than that of the corresponding MHV complex, which may explain the different inhibitory potencies of the HR2 peptides. Analogous to other class I viral fusion proteins, the six-helix complex supposedly represents a postfusion conformation that is formed after insertion of the fusion peptide, proposed here for coronaviruses to be located immediately upstream of HR1, into the target membrane. The resulting close apposition of fusion peptide and spike transmembrane domain facilitates membrane fusion. The inhibitory potency of the SARS-CoV HR2-peptides provides an attractive basis for the development of a therapeutic drug for SARS.