Analysis of the avian coronavirus spike protein reveals heterogeneity in the glycans present.
Analysis of the avian coronavirus spike protein reveals heterogeneity in the glycans present.
复制标题
对禽冠状病毒尖峰蛋白的分析揭示了存在的聚糖中的异质性。
DOI：
10.1099/jgv.0.001642
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发表时间：
2021-08
期刊：
影响因子：
--
通讯作者：
Bickerton E
中科院分区：
文献类型：
--
作者：
Stevenson-Leggett P;Armstrong S;Keep S;Britton P;Bickerton E
传染性支气管炎病毒（IBV）是一种具有重要经济意义的冠状病毒，作为传染性支气管炎的病原体，给全球家禽业造成了破坏性损失。冠状病毒刺突（S）糖蛋白是从病毒体表面突出的大的I型膜蛋白，其促进附着和进入宿主细胞。IBV S蛋白被切割成两个亚基，S1和S2，其中后者已被鉴定为细胞嗜性的决定因素。最近的研究表明，在哺乳动物和昆虫细胞中表达冠状病毒S蛋白的蛋白质表面具有高水平的糖基化。在这里，我们使用鸡胚蛋中繁殖的IBV来探索来自天然宿主鸡细胞感染的病毒的聚糖谱。我们确定了蛋白质表面上的多种聚糖类型，并发现了一种菌株特异性的依赖于复杂的聚糖，用于在体外通过单克隆抗体识别S2亚基，在化学抑制复杂的糖基化后对病毒复制没有影响。单克隆或多克隆抗体的病毒中和作用不受影响。在分析了四种IBV毒株S蛋白的预测糖基化位点后，我们通过质谱法证实了致病性实验室毒株M41-CK在18个位点的糖基化。进一步表征揭示了这些位点中的六个位点存在的聚糖之间的异质性，表明IBV病毒粒子上单个S蛋白的聚糖谱存在差异。这些结果证明了复合聚糖在IBV复制中的非特异性作用，表明其参与抗体识别但不参与中和。
Infectious bronchitis virus (IBV) is an economically important coronavirus, causing damaging losses to the poultry industry worldwide as the causative agent of infectious bronchitis. The coronavirus spike (S) glycoprotein is a large type I membrane protein protruding from the surface of the virion, which facilitates attachment and entry into host cells. The IBV S protein is cleaved into two subunits, S1 and S2, the latter of which has been identified as a determinant of cellular tropism. Recent studies expressing coronavirus S proteins in mammalian and insect cells have identified a high level of glycosylation on the protein’s surface. Here we used IBV propagated in embryonated hens’ eggs to explore the glycan profile of viruses derived from infection in cells of the natural host, chickens. We identified multiple glycan types on the surface of the protein and found a strain-specific dependence on complex glycans for recognition of the S2 subunit by a monoclonal antibody in vitro, with no effect on viral replication following the chemical inhibition of complex glycosylation. Virus neutralization by monoclonal or polyclonal antibodies was not affected. Following analysis of predicted glycosylation sites for the S protein of four IBV strains, we confirmed glycosylation at 18 sites by mass spectrometry for the pathogenic laboratory strain M41-CK. Further characterization revealed heterogeneity among the glycans present at six of these sites, indicating a difference in the glycan profile of individual S proteins on the IBV virion. These results demonstrate a non-specific role for complex glycans in IBV replication, with an indication of an involvement in antibody recognition but not neutralisation.
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影响因子：
4.1
作者：
Bande F;Arshad SS;Bejo MH;Moeini H;Omar AR
通讯作者：
Omar AR
影响因子：
5
作者：
Andoh K;Suenaga K;Sakaguchi M;Yamazaki K;Honda T
通讯作者：
Honda T
影响因子：
3.7
作者：
de Vries, Robert P.;de Vries, Erik;de Haan, Cornelis A. M.
通讯作者：
de Haan, Cornelis A. M.
影响因子：
2.7
作者：
COOK, JKA;DARBYSHIRE, JH;PETERS, RW
通讯作者：
PETERS, RW
影响因子：
5.4
作者：
Bickerton E;Maier HJ;Stevenson-Leggett P;Armesto M;Britton P
通讯作者：
Britton P