TY - JOUR T1 - Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins JF - bioRxiv DO - 10.1101/2020.03.28.013276 SP - 2020.03.28.013276 AU - Yong Zhang AU - Wanjun Zhao AU - Yonghong Mao AU - Yaohui Chen AU - Shisheng Wang AU - Yi Zhong AU - Tao Su AU - Meng Gong AU - Dan Du AU - Xiaofeng Lu AU - Jingqiu Cheng AU - Hao Yang Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/06/30/2020.03.28.013276.abstract N2 - The glycoprotein spike (S) on the surface of 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 subunits. 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, primarily 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. 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.Competing Interest StatementThe authors have declared no competing interest.ACE2angiotensin-converting enzyme IICryo-EMcryoelectron microscopyEenvelope proteinHCoV-NL63human coronavirus NL63Mmembrane proteinMSmass spectrometryMERS-CoVMiddle East respiratory syndrome coronavirusNnucleocapsid proteinRBDreceptor-binding domainSspike proteinSARS-CoV-2severe acute respiratory syndrome coronavirusSCEstepped collision energyZic-HILICzwitterionic hydrophilic interaction liquid chromatography ER -