Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins.
Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins.
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DOI：
10.1074/mcp.ra120.002295
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Yang H
中科院分区：
文献类型：
--
作者：
Zhang Y;Zhao W;Mao Y;Chen Y;Wang S;Zhong Y;Su T;Gong M;Du D;Lu X;Cheng J;Yang H
The glycoprotein spike (S) on the surface of severe acute respiratory syndrome coronavirus (SARS-CoV-2) is a determinant for viral invasion and host immune response. Herein, we characterized the site-specific N-glycosylation of S protein at the level of intact glycopeptides. All 22 potential N-glycosites were identified in the S-protein protomer and were found to be preserved among the 753 SARS-CoV-2 genome sequences. The glycosites exhibited glycoform heterogeneity as expected for a human cell-expressed protein subunit. We identified masses that correspond to 157 N-glycans, primarily of the complex type. In contrast, the insect cell-expressed S protein contained 38 N-glycans, completely of the high-mannose type. Our results revealed that the glycan types were highly determined by the differential processing of N-glycans among human and insect cells, regardless of the glycosites’ location. Moreover, the N-glycan compositions were conserved among different sizes of subunits. Our study indicates that the S protein N-glycosylation occurs regularly at each site, albeit the occupied N-glycans were diverse and heterogenous. This N-glycosylation landscape and the differential N-glycan patterns among distinct host cells are expected to shed light on the infection mechanism and present a positive view for the development of vaccines and targeted drugs. Decoding N-glycosites and site-specific N-glycans on spike protein The variation of N-glycosylation sequons among 753 SARS-CoV-2 variants N-glycosylation comparison among different lengths of spike proteins from different hosts Prospective insights on host immunity and vaccine development Site-specific N-glycosylation of SARS-CoV-2 spike proteins expressed in human and insect cells has been decoded via enriched glycopeptides using a stepped collision energy HCD scan. The regular N-glycan occupancy on preserved N-glycosites has been revealed, despite the heterogeneity in N-glycan compositions. The complex N-glycans on the human cell-expressed spike protein reflect the relatively accessible antigens and epitopes for immune recognition. The insect cell-expressed spike protein subunits decorated with immature high-mannose N-glycans might become promising recombinant vaccine candidates.
