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Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration.
Elife. 2020 10 26; 9E

Abstract

The Spike protein of SARS-CoV-2, its receptor-binding domain (RBD), and its primary receptor ACE2 are extensively glycosylated. The impact of this post-translational modification on viral entry is yet unestablished. We expressed different glycoforms of the Spike-protein and ACE2 in CRISPR-Cas9 glycoengineered cells, and developed corresponding SARS-CoV-2 pseudovirus. We observed that N- and O-glycans had only minor contribution to Spike-ACE2 binding. However, these carbohydrates played a major role in regulating viral entry. Blocking N-glycan biosynthesis at the oligomannose stage using both genetic approaches and the small molecule kifunensine dramatically reduced viral entry into ACE2 expressing HEK293T cells. Blocking O-glycan elaboration also partially blocked viral entry. Mechanistic studies suggest multiple roles for glycans during viral entry. Among them, inhibition of N-glycan biosynthesis enhanced Spike-protein proteolysis. This could reduce RBD presentation on virus, lowering binding to host ACE2 and decreasing viral entry. Overall, chemical inhibitors of glycosylation may be evaluated for COVID-19.

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Authors+Show Affiliations

Chemical & Biological Engineering, State University of New York, Buffalo, United States.

Chemical & Biological Engineering, State University of New York, Buffalo, United States.

Chemical & Biological Engineering, State University of New York, Buffalo, United States.

Chemical & Biological Engineering, State University of New York, Buffalo, United States.

Chemical & Biological Engineering, State University of New York, Buffalo, United States.

Chemical & Biological Engineering, State University of New York, Buffalo, United States.

Biomedical Engineering, State University of New York, Buffalo, United States.

Chemical & Biological Engineering, State University of New York, Buffalo, United States. Biomedical Engineering, State University of New York, Buffalo, United States.

Chemical & Biological Engineering, State University of New York, Buffalo, United States. Biomedical Engineering, State University of New York, Buffalo, United States. Medicine, State University of New York, Buffalo, United States. Clinical & Translational Research Center, Buffalo, United States.

MeSH

AlkaloidsAngiotensin-Converting Enzyme 2Gene Knockout TechniquesGlycosylationHEK293 CellsHost Microbial InteractionsHumansMass SpectrometryMolecular Dynamics SimulationPolysaccharidesProtein BindingProtein Processing, Post-TranslationalReceptors, VirusSARS-CoV-2Spike Glycoprotein, CoronavirusVirus Internalization

Pub Type(s)

Journal Article

Research Support, N.I.H., Extramural

Language

eng

PubMed ID

33103998

Citation

Yang, Qi, et al. "Inhibition of SARS-CoV-2 Viral Entry Upon Blocking N- and O-glycan Elaboration." ELife, vol. 9, 2020.

Yang Q, Hughes TA, Kelkar A, et al. Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration. Elife. 2020;9.

Yang, Q., Hughes, T. A., Kelkar, A., Yu, X., Cheng, K., Park, S., Huang, W. C., Lovell, J. F., & Neelamegham, S. (2020). Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration. ELife, 9. https://doi.org/10.7554/eLife.61552

Yang Q, et al. Inhibition of SARS-CoV-2 Viral Entry Upon Blocking N- and O-glycan Elaboration. Elife. 2020 10 26;9 PubMed PMID: 33103998.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration. AU - Yang,Qi, AU - Hughes,Thomas A, AU - Kelkar,Anju, AU - Yu,Xinheng, AU - Cheng,Kai, AU - Park,Sheldon, AU - Huang,Wei-Chiao, AU - Lovell,Jonathan F, AU - Neelamegham,Sriram, Y1 - 2020/10/26/ PY - 2020/07/29/received PY - 2020/10/24/accepted PY - 2020/10/27/pubmed PY - 2020/12/15/medline PY - 2020/10/26/entrez KW - COVID-19 KW - SARS-CoV-2 KW - biochemistry KW - chemical biology KW - furin KW - glycoscience KW - human KW - infectious disease KW - kifunensine KW - microbiology KW - spike KW - virus JF - eLife JO - Elife VL - 9 N2 - The Spike protein of SARS-CoV-2, its receptor-binding domain (RBD), and its primary receptor ACE2 are extensively glycosylated. The impact of this post-translational modification on viral entry is yet unestablished. We expressed different glycoforms of the Spike-protein and ACE2 in CRISPR-Cas9 glycoengineered cells, and developed corresponding SARS-CoV-2 pseudovirus. We observed that N- and O-glycans had only minor contribution to Spike-ACE2 binding. However, these carbohydrates played a major role in regulating viral entry. Blocking N-glycan biosynthesis at the oligomannose stage using both genetic approaches and the small molecule kifunensine dramatically reduced viral entry into ACE2 expressing HEK293T cells. Blocking O-glycan elaboration also partially blocked viral entry. Mechanistic studies suggest multiple roles for glycans during viral entry. Among them, inhibition of N-glycan biosynthesis enhanced Spike-protein proteolysis. This could reduce RBD presentation on virus, lowering binding to host ACE2 and decreasing viral entry. Overall, chemical inhibitors of glycosylation may be evaluated for COVID-19. SN - 2050-084X UR - https://www.unboundmedicine.com/medline/citation/33103998/Inhibition_of_SARS_CoV_2_viral_entry_upon_blocking_N__and_O_glycan_elaboration_ DB - PRIME DP - Unbound Medicine ER -