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Analysis of glycosylation and disulfide bonding of wild-type SARS-CoV-2 spike glycoprotein

Shijian Zhang, Eden P. Go, Haitao Ding, Saumya Anang, John C. Kappes, Heather Desaire, View ORCID ProfileJoseph Sodroski
doi: https://doi.org/10.1101/2021.04.01.438120
Shijian Zhang
1Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Department of Microbiology, Harvard Medical School, Boston, MA 02215, USA
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Eden P. Go
2Department of Chemistry, University of Kansas, Lawrence, KS 66049, USA
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Haitao Ding
3Department of Medicine, University of Alabama at Birmingham, AL 35294, USA
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Saumya Anang
1Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Department of Microbiology, Harvard Medical School, Boston, MA 02215, USA
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John C. Kappes
3Department of Medicine, University of Alabama at Birmingham, AL 35294, USA
4Birmingham Veterans Affairs Medical Center, Research Service, Birmingham, AL 35233, USA
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Heather Desaire
2Department of Chemistry, University of Kansas, Lawrence, KS 66049, USA
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Joseph Sodroski
1Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Department of Microbiology, Harvard Medical School, Boston, MA 02215, USA
5Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, MA 02215, USA
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  • ORCID record for Joseph Sodroski
  • For correspondence: joseph_sodroski@dfci.harvard.edu
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ABSTRACT

The SARS-CoV-2 coronavirus, the etiologic agent of COVID-19, uses its spike (S) glycoprotein anchored in the viral membrane to enter host cells. The S glycoprotein is the major target for neutralizing antibodies elicited by natural infection and by vaccines. Approximately 35% of the SARS-CoV-2 S glycoprotein consists of carbohydrate, which can influence virus infectivity and susceptibility to antibody inhibition. We found that virus-like particles produced by coexpression of SARS-CoV-2 S, M, E and N proteins contained spike glycoproteins that were extensively modified by complex carbohydrates. We used a fucose-selective lectin to enrich the Golgi-resident fraction of a wild-type SARS-CoV-2 S glycoprotein trimer, and determined its glycosylation and disulfide bond profile. Compared with soluble or solubilized S glycoproteins modified to prevent proteolytic cleavage and to retain a prefusion conformation, more of the wild-type S glycoprotein N-linked glycans are processed to complex forms. Even Asn 234, a significant percentage of which is decorated by high-mannose glycans on soluble and virion S trimers, is predominantly modified in the Golgi by processed glycans. Three incompletely occupied sites of O-linked glycosylation were detected. Viruses pseudotyped with natural variants of the serine/threonine residues implicated in O-linked glycosylation were generally infectious and exhibited sensitivity to neutralization by soluble ACE2 and convalescent antisera comparable to that of the wild-type virus. Unlike other natural cysteine variants, a Cys15Phe (C15F) mutant retained partial, but unstable, infectivity. These findings enhance our understanding of the Golgi processing of the native SARS-CoV-2 S glycoprotein carbohydrates and could assist the design of interventions.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted April 01, 2021.
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Analysis of glycosylation and disulfide bonding of wild-type SARS-CoV-2 spike glycoprotein
Shijian Zhang, Eden P. Go, Haitao Ding, Saumya Anang, John C. Kappes, Heather Desaire, Joseph Sodroski
bioRxiv 2021.04.01.438120; doi: https://doi.org/10.1101/2021.04.01.438120
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Analysis of glycosylation and disulfide bonding of wild-type SARS-CoV-2 spike glycoprotein
Shijian Zhang, Eden P. Go, Haitao Ding, Saumya Anang, John C. Kappes, Heather Desaire, Joseph Sodroski
bioRxiv 2021.04.01.438120; doi: https://doi.org/10.1101/2021.04.01.438120

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