Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins using High-Resolution Mass Spectrometry

Yong Zhang; Wanjun Zhao; Yonghong Mao; Shisheng Wang; Yi Zhong; Tao Su; Meng Gong; Xiaofeng Lu; Jingqiu Cheng; Hao Yang

. CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint unambiguously assigned N-glycosite using trypsin digestion; Green arrow: unambiguously assigned N-glycosite using Glu-C digestion; Yellow arrow: unambiguously assigned N-glycosite using the combination of trypsin and Glu-C digestion. The unambiguously glycosite was determined by at least twice identification within each digestion list in Table S1 and Table S2 . The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint Table S1 . Glycoproteomic identification of the N-glycosites on recombinant SARS-CoV-2 S protein expressed in insect cells. Table S2 . Glycoproteomic identification of the N-glycosites on recombinant SARS-CoV-2 S protein expressed in human cells. Table S3 . Site-specific N-glycosylation characterization of recombinant SARS-CoV-2 spike protein expressed in insect cells. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint Table S4 . Site-specific N-glycosylation characterization of recombinant SARS-CoV-2 spike protein expressed in human cells. Table S5 . Mutation frequency of SARS-CoV-2 spike protein. Table S6 . Intact N-glycopeptides of the recombinant RBD of the SARS-CoV-2 spike protein. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint N122 N149 N165 . CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint N234 N282 N331 . CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint N343 N603 N657 . CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint . CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint Supplementary Figure S8 . Microheterogeneity and macroheterogeneity of the N-linked glycopeptides of the S protein.