使用仿生 Trp-Arg 材料对 SARS-CoV-2 尖峰蛋白进行综合 O-糖基化分析,Analytical Chemistry

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使用仿生 Trp-Arg 材料对 SARS-CoV-2 尖峰蛋白进行综合 O-糖基化分析
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-07-20 , DOI: 10.1021/acs.analchem.0c04634
Xuefang Dong ₁ , Cheng Chen ₁ , Jingyu Yan ₁ , Xiaofei Zhang ₁ , Xiuling Li ₁ , Xinmiao Liang ₁

Affiliation

由严重急性呼吸系统综合症冠状病毒-2 (SARS-CoV-2) 引起的 COVID-19 大流行是严重的公共卫生威胁。大多数针对 SARS-CoV-2 的疫苗都针对高度糖基化的刺突蛋白 (S)。充分了解该蛋白质的糖基化特征是成功开发疫苗的关键。与 SARS-CoV-2 S 上已确认的 22 个 N-糖基化位点不同，仅报道了该蛋白质上的几个 O-糖基化位点。这种差异主要归因于 O-糖基化的极低化学计量。在此，我们设计了仿生材料，Trp-Arg (WR) 单体接枝二氧化硅微球（命名为 WR-SiO ₂），这些仿生材料可以高选择性地富集 N 和 O 连接的糖肽。和WR-SiO ₂在O-连接糖肽富集过程中，可以抵抗非糖肽对100摩尔倍BSA的干扰。我们利用 WR-SiO ₂全面分析了重组 SARS-CoV-2 S 的 O-糖基化特征。在 SARS-CoV-2 S 上鉴定了 27 个 O-糖基化位点，包括 18 个明确的位点。我们的研究表明，仿生聚合物可以为 O-连接的糖肽提供特定的选择性，为生物领域的 O-糖基化研究铺平道路。除了 SARS-CoV-2 S 的 N-糖基化外，SARS-CoV-2 S 的 O-糖基化特征可能补充了全面的糖基化。

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Comprehensive O-Glycosylation Analysis of the SARS-CoV-2 Spike Protein with Biomimetic Trp-Arg Materials

The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious public health threat. Most vaccines against SARS-CoV-2 target the highly glycosylated spike protein (S). A good knowledge of the glycosylation profile of this protein is key to successful vaccine development. Unlike the 22 confirmed N-glycosylation sites on SARS-CoV-2 S, only a few O-glycosylation sites on this protein have been reported. This difference is mainly ascribed to the extremely low stoichiometry of O-glycosylation. Herein, we designed the biomimetic materials, Trp-Arg (WR) monomer-grafted silica microspheres (designated as WR-SiO₂), and these biomimetic materials can enrich N- and O-linked glycopeptides with high selectivity. And WR-SiO₂ can resist the nonglycopeptides’ interference with the 100 molar fold of BSA during O-linked glycopeptide enrichment. We utilized WR-SiO₂ to comprehensively analyze the O-glycosylation profile of recombinant SARS-CoV-2 S. Twenty-seven O-glycosylation sites including 18 unambiguous sites are identified on SARS-CoV-2 S. Our study demonstrates that the biomimetic polymer can offer specific selectivity for O-linked glycopeptides and pave the way for O-glycosylation research in biological fields. The O-glycosylation profile of SARS-CoV-2 S might supplement the comprehensive glycosylation in addition to N-glycosylation of SARS-CoV-2 S.

更新日期：2021-08-03

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