Combinatorial Approach with Mass Spectrometry and Lectin Microarray Dissected Site-Specific Glycostem and Glycoleaf Features of the Virion-Derived Spike Protein of Ancestral and γ Variant SARS-CoV-2 Strains

作     者：Hiono, Takahiro Sakaue, Hiroaki Tomioka, Azusa Kaji, Hiroyuki Sasaki, Michihito Orba, Yasuko Sawa, Hirofumi Kuno, Atsushi 

作者机构：Natl Inst Adv Ind Sci & Technol Cellular & Mol Biotechnol Res Inst Mol & Cellular Glycoprote Res Grp Tsukuba Ibaraki 3058565 Japan Hokkaido Univ One Hlth Res Ctr Sapporo Hokkaido 0600818 Japan Hokkaido Univ Fac Vet Med Dept Dis Control Lab Microbiol Sapporo Hokkaido 0600818 Japan Hokkaido Univ Int Inst Zoonosis Control Int Collaborat Unit Sapporo Hokkaido 0010020 Japan Hokkaido Univ Inst Vaccine Res & Dev HU IVReD Sapporo Hokkaido 0010021 Japan Hokkaido Univ Int Inst Zoonosis Control Div Mol Pathobiol Sapporo Hokkaido 0010020 Japan 

出 版 物：《JOURNAL OF PROTEOME RESEARCH》 (蛋白质组研究杂志)

年 卷 期：2024年第23卷第4期

页      面：1408-1419页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 

基　　金：Japan Agency for Medical Research and Development [18K15176, 21K14982] Japan Society for the Promotion of Science (JSPS) KAKENHI [16809263, 20he0522002j0001, JP223fa627005] Japan Agency for Medical Research and Development (AMED) 

主　　题：SARS-CoV-2 COVID-19 spike protein glycosylation glycoproteomics lectin microarray mass spectrometry 

摘      要：The coronavirus disease (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has impacted public health globally. As the glycosylation of viral envelope glycoproteins is strongly associated with their immunogenicity, intensive studies have been conducted on the glycans of the glycoprotein of SARS-CoV-2, the spike (S) protein. Here, we conducted intensive glycoproteomic analyses of the SARS-CoV-2 S protein of ancestral and gamma-variant strains using a combinatorial approach with two different technologies: mass spectrometry (MS) and lectin microarrays (LMA). Our unique MS1-based glycoproteomic technique, Glyco-RIDGE, in addition to MS2-based Byonic search, identified 1448 (ancestral strain) and 1785 (gamma-variant strain) site-specific glycan compositions, respectively. Asparagine at amino acid position 20 (N20) is mainly glycosylated within two successive potential glycosylation sites, N17 and N20, of the gamma-variant S protein;however, we found low-frequency glycosylation at N17. Our novel approaches, glycostem mapping and glycoleaf scoring, also illustrate the moderately branched/extended, highly fucosylated, and less sialylated natures of the glycoforms of S proteins. Subsequent LMA analysis emphasized the intensive end-capping of glycans by Lewis fucoses, which complemented the glycoproteomic features. These results illustrate the high-resolution glycoproteomic features of the SARS-CoV-2 S protein, contributing to vaccine design and understanding of viral protein synthesis.