Spike Protein, S, of Human Coronavirus HKU1: Role in Viral Life Cycle and Application in Antibody Detection (Retracted Article)
Spike Protein, S, of Human Coronavirus HKU1: Role in Viral Life Cycle and Application in Antibody Detection (Retracted Article)
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DOI：
10.3181/0806-rm-197
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发表时间：
2008-12-01
影响因子：
3.2
通讯作者：
Yuen, Kwok-Yung
中科院分区：
文献类型：
--
作者：
Chan, Che-Man;Woo, Patrick C. Y.;Yuen, Kwok-Yung
我们最近描述了一种新的全球人类呼吸道病毒的发现，基因组，临床特征，基因型和进化，命名为人类冠状病毒HKU1 (HCoV-HKU1)，尚未培养。我们通过塞姆利基森林病毒（SFV）系统表达了一个c端flag标记的CoV-HKU1刺突(S)蛋白，并研究了其成熟谱。脉冲追踪标记显示S-FLAG表达为单体和三聚体的高甘露糖n聚糖。成熟过程中主要裂解为S1和S2亚结构域。S1被分泌到培养基中。免疫荧光分析显示S沿着从内质网到质膜的分泌途径。呋喃或类呋喃酶抑制剂抑制了S的裂解和HCoV-HKU1 S伪病毒的释放。以细胞为基础表达的全长S-FLAG可被HCoV-HKU1型肺炎患者恢复期血清识别。这些数据表明，在我们的系统中表达的HCoV-HKU1刺突的天然形式可用于开发血清学诊断检测和了解S在病毒生命周期中的作用。中国生物医学工程学报（英文版），2008
We recently described the discovery, genome, clinical features, genotypes and evolution of a novel and global human respiratory virus named human coronavirus HKU1 (HCoV-HKU1) which is not yet culturable. We expressed a C-terminal FLAG-tagged CoV-HKU1 spike (S) protein by the Semliki Forest Virus (SFV) system and investigated its maturation profile. Pulse chase labeling revealed that S-FLAG was expressed as high-mannose N-glycans of monomers and trimers. It was predominantly cleaved into subdomains S1 and S2 during maturation. S1 was secreted into the medium. Immunofluorescence analysis visualized S along the secretory pathway from endoplasmic reticulum to plasma membrane. Cleavage of S and release of HCoV-HKU1 S pseudotyped virus were inhibited by furin or furin-like enzyme inhibitors. The cell-based expressed full-length S-FLAG could be recognized by the convalescent serum obtained from a patient with HCoV-HKU1 pneumonia. The data suggest that the native form of HCoV-HKU1 spike expressed in our system can be used in developing serological diagnostic assay and in understanding the role of S in the viral life cycle. Exp Biol Med 233:1527-1536, 2008