SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity
ABSTRACT The glycosylation of viral envelope proteins can play important roles in virus biology and immune evasion. The spike (S) glycoprotein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) includes 22 N-linked glycosylation sequons and 17 O-linked glycosites. We investigated the ef...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2024-02-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.01672-23 |
| _version_ | 1827150268542746624 |
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| author | Fengwen Zhang Fabian Schmidt Frauke Muecksch Zijun Wang Anna Gazumyan Michel C. Nussenzweig Christian Gaebler Marina Caskey Theodora Hatziioannou Paul D. Bieniasz |
| author_facet | Fengwen Zhang Fabian Schmidt Frauke Muecksch Zijun Wang Anna Gazumyan Michel C. Nussenzweig Christian Gaebler Marina Caskey Theodora Hatziioannou Paul D. Bieniasz |
| author_sort | Fengwen Zhang |
| collection | DOAJ |
| description | ABSTRACT The glycosylation of viral envelope proteins can play important roles in virus biology and immune evasion. The spike (S) glycoprotein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) includes 22 N-linked glycosylation sequons and 17 O-linked glycosites. We investigated the effect of individual glycosylation sites on SARS-CoV-2 S function in pseudotyped virus infection assays and on sensitivity to monoclonal and polyclonal neutralizing antibodies. In most cases, the removal of individual glycosylation sites decreased the infectiousness of the pseudotyped virus. For glycosylation mutants in the N-terminal domain and the receptor-binding domain (RBD), reduction in pseudotype infectivity was predicted by a commensurate reduction in the level of virion-incorporated S protein and reduced S trafficking to the cell surface. Notably, the presence of a glycan at position N343 within the RBD had diverse effects on neutralization by RBD-specific monoclonal antibodies cloned from convalescent individuals. The N343 glycan reduced the overall sensitivity to polyclonal antibodies in plasma from COVID-19 convalescent individuals, suggesting a role for SARS-CoV-2 S glycosylation in immune evasion. However, vaccination of convalescent individuals produced neutralizing activity that was resilient to the inhibitory effect of the N343 glycan.IMPORTANCEThe attachment of glycans to the spike proteins of viruses during their synthesis and movement through the secretory pathway can affect their properties. This study shows that the glycans attached to the severe acute respiratory syndrome coronavirus-2 spike protein enable its movement to the cell surface and incorporation into virus particles. Certain glycans, including one that is attached to asparagine 343 in the receptor-binding domain of the spike protein, can also affect virus neutralization by antibodies. This glycan can increase or decrease sensitivity to individual antibodies, likely through direct effects on antibody epitopes and modulation of spike conformation. However, the overall effect of the glycan in the context of the polyclonal mixture of antibodies in convalescent serum is to reduce neutralization sensitivity. Overall, this study highlights the complex effects of glycosylation on spike protein function and immune evasion. |
| first_indexed | 2024-03-08T01:26:13Z |
| format | Article |
| id | doaj.art-d4d51df98c9c47238de71504346cf0f1 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2025-03-20T21:31:23Z |
| publishDate | 2024-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-d4d51df98c9c47238de71504346cf0f12024-08-11T18:24:59ZengAmerican Society for MicrobiologymBio2150-75112024-02-0115210.1128/mbio.01672-23SARS-CoV-2 spike glycosylation affects function and neutralization sensitivityFengwen Zhang0Fabian Schmidt1Frauke Muecksch2Zijun Wang3Anna Gazumyan4Michel C. Nussenzweig5Christian Gaebler6Marina Caskey7Theodora Hatziioannou8Paul D. Bieniasz9Laboratory of Retrovirology, The Rockefeller University, New York, New York, USALaboratory of Retrovirology, The Rockefeller University, New York, New York, USALaboratory of Retrovirology, The Rockefeller University, New York, New York, USALaboratory of Molecular Immunology, The Rockefeller University, New York, New York, USALaboratory of Molecular Immunology, The Rockefeller University, New York, New York, USALaboratory of Molecular Immunology, The Rockefeller University, New York, New York, USALaboratory of Molecular Immunology, The Rockefeller University, New York, New York, USALaboratory of Molecular Immunology, The Rockefeller University, New York, New York, USALaboratory of Retrovirology, The Rockefeller University, New York, New York, USALaboratory of Retrovirology, The Rockefeller University, New York, New York, USAABSTRACT The glycosylation of viral envelope proteins can play important roles in virus biology and immune evasion. The spike (S) glycoprotein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) includes 22 N-linked glycosylation sequons and 17 O-linked glycosites. We investigated the effect of individual glycosylation sites on SARS-CoV-2 S function in pseudotyped virus infection assays and on sensitivity to monoclonal and polyclonal neutralizing antibodies. In most cases, the removal of individual glycosylation sites decreased the infectiousness of the pseudotyped virus. For glycosylation mutants in the N-terminal domain and the receptor-binding domain (RBD), reduction in pseudotype infectivity was predicted by a commensurate reduction in the level of virion-incorporated S protein and reduced S trafficking to the cell surface. Notably, the presence of a glycan at position N343 within the RBD had diverse effects on neutralization by RBD-specific monoclonal antibodies cloned from convalescent individuals. The N343 glycan reduced the overall sensitivity to polyclonal antibodies in plasma from COVID-19 convalescent individuals, suggesting a role for SARS-CoV-2 S glycosylation in immune evasion. However, vaccination of convalescent individuals produced neutralizing activity that was resilient to the inhibitory effect of the N343 glycan.IMPORTANCEThe attachment of glycans to the spike proteins of viruses during their synthesis and movement through the secretory pathway can affect their properties. This study shows that the glycans attached to the severe acute respiratory syndrome coronavirus-2 spike protein enable its movement to the cell surface and incorporation into virus particles. Certain glycans, including one that is attached to asparagine 343 in the receptor-binding domain of the spike protein, can also affect virus neutralization by antibodies. This glycan can increase or decrease sensitivity to individual antibodies, likely through direct effects on antibody epitopes and modulation of spike conformation. However, the overall effect of the glycan in the context of the polyclonal mixture of antibodies in convalescent serum is to reduce neutralization sensitivity. Overall, this study highlights the complex effects of glycosylation on spike protein function and immune evasion.https://journals.asm.org/doi/10.1128/mbio.01672-23SARS-CoV-2antibody functionneutralizationglycosylation |
| spellingShingle | Fengwen Zhang Fabian Schmidt Frauke Muecksch Zijun Wang Anna Gazumyan Michel C. Nussenzweig Christian Gaebler Marina Caskey Theodora Hatziioannou Paul D. Bieniasz SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity mBio SARS-CoV-2 antibody function neutralization glycosylation |
| title | SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity |
| title_full | SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity |
| title_fullStr | SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity |
| title_full_unstemmed | SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity |
| title_short | SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity |
| title_sort | sars cov 2 spike glycosylation affects function and neutralization sensitivity |
| topic | SARS-CoV-2 antibody function neutralization glycosylation |
| url | https://journals.asm.org/doi/10.1128/mbio.01672-23 |
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