Deducing the N- and O-glycosylation profile of the spike protein of novel coronavirus SARS-CoV-2
Deducing the N- and O-glycosylation profile of the spike protein of novel coronavirus SARS-CoV-2
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DOI：
10.1093/glycob/cwaa042
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发表时间：
2020-12-01
期刊：
影响因子：
4.3
通讯作者：
Azadi, Parastoo
中科院分区：
文献类型：
--
作者：
Shajahan, Asif;Supekar, Nitin T.;Azadi, Parastoo
目前出现了由严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)引起的新型冠状病毒大流行，这要求开发新的治疗策略来防止死亡率的快速增长。冠状病毒刺突蛋白(S)是一种高度糖基化的蛋白，它促进病毒的附着、进入和膜融合，在诱导宿主免疫反应中起着关键作用。Spike蛋白由两个蛋白质亚基(S1和S2)组成，这两个亚基总共具有22个潜在的N-糖基化位点。在此，我们报道了通过高分辨质谱学对人类细胞表达的S蛋白亚基S1和S2的糖基化图谱。我们已经表征了Spike蛋白上的定量N-糖基化图谱，有趣的是，我们观察到了Spike蛋白亚单位S1受体结合域上意外的O-糖基化修饰。尽管已经预测了SARS-CoV-2刺突蛋白上的O-糖基化，但这是第一次关于O-糖基化的位置和连接在亚基S1上的O-糖链的特性的实验数据的报告。除了使用生物信息学工具来确认Spike蛋白中糖基化的复杂性外，我们关于N-和O-糖基化的数据还通过对每个糖肽谱的广泛手动解释而得到加强。对Spike蛋白上的糖链谱系的阐明为病毒结合研究提供了见解，更重要的是，推动了对合适疫苗候选的研究。
The current emergence of the novel coronavirus pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) demands the development of new therapeutic strategies to prevent rapid progress of mortalities. The coronavirus spike (S) protein, which facilitates viral attachment, entry and membrane fusion is heavily glycosylated and plays a critical role in the elicitation of the host immune response. The spike protein is comprised of two protein subunits (S1 and S2), which together possess 22 potential N-glycosylation sites. Herein, we report the glycosylation mapping on spike protein subunits S1 and S2 expressed on human cells through high-resolution mass spectrometry. We have characterized the quantitative N-glycosylation profile on spike protein and interestingly, observed unexpected O-glycosylation modifications on the receptor-binding domain of spike protein subunit S1. Even though O-glycosylation has been predicted on the spike protein of SARS-CoV-2, this is the first report of experimental data for both the site of O-glycosylation and identity of the O-glycans attached on the subunit S1. Our data on the N-and O-glycosylation are strengthened by extensive manual interpretation of each glycopeptide spectra in addition to using bioinformatics tools to confirm the complexity of glycosylation in the spike protein. The elucidation of the glycan repertoire on the spike protein provides insights into the viral binding studies and more importantly, propels research toward the development of a suitable vaccine candidate.