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Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells
Enveloped viruses hijack not only the host translation processes, but also its glycosylation machinery, and to a variable extent cover viral surface proteins with tolerogenic host-like structures. SARS-CoV-2 surface protein S presents as a trimer on the viral surface and is covered by a dense shield...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064498/ https://www.ncbi.nlm.nih.gov/pubmed/33806155 http://dx.doi.org/10.3390/v13040551 |
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author | Bagdonaite, Ieva Thompson, Andrew J. Wang, Xiaoning Søgaard, Max Fougeroux, Cyrielle Frank, Martin Diedrich, Jolene K. Yates, John R. Salanti, Ali Vakhrushev, Sergey Y. Paulson, James C. Wandall, Hans H. |
author_facet | Bagdonaite, Ieva Thompson, Andrew J. Wang, Xiaoning Søgaard, Max Fougeroux, Cyrielle Frank, Martin Diedrich, Jolene K. Yates, John R. Salanti, Ali Vakhrushev, Sergey Y. Paulson, James C. Wandall, Hans H. |
author_sort | Bagdonaite, Ieva |
collection | PubMed |
description | Enveloped viruses hijack not only the host translation processes, but also its glycosylation machinery, and to a variable extent cover viral surface proteins with tolerogenic host-like structures. SARS-CoV-2 surface protein S presents as a trimer on the viral surface and is covered by a dense shield of N-linked glycans, and a few O-glycosites have been reported. The location of O-glycans is controlled by a large family of initiating enzymes with variable expression in cells and tissues and hence is difficult to predict. Here, we used our well-established O-glycoproteomic workflows to map the precise positions of O-linked glycosylation sites on three different entities of protein S—insect cell or human cell-produced ectodomains, or insect cell derived receptor binding domain (RBD). In total 25 O-glycosites were identified, with similar patterns in the two ectodomains of different cell origin, and a distinct pattern of the monomeric RBD. Strikingly, 16 out of 25 O-glycosites were located within three amino acids from known N-glycosites. However, O-glycosylation was primarily found on peptides that were unoccupied by N-glycans, and otherwise had low overall occupancy. This suggests possible complementary functions of O-glycans in immune shielding and negligible effects of O-glycosylation on subunit vaccine design for SARS-CoV-2. |
format | Online Article Text |
id | pubmed-8064498 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-80644982021-04-24 Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells Bagdonaite, Ieva Thompson, Andrew J. Wang, Xiaoning Søgaard, Max Fougeroux, Cyrielle Frank, Martin Diedrich, Jolene K. Yates, John R. Salanti, Ali Vakhrushev, Sergey Y. Paulson, James C. Wandall, Hans H. Viruses Article Enveloped viruses hijack not only the host translation processes, but also its glycosylation machinery, and to a variable extent cover viral surface proteins with tolerogenic host-like structures. SARS-CoV-2 surface protein S presents as a trimer on the viral surface and is covered by a dense shield of N-linked glycans, and a few O-glycosites have been reported. The location of O-glycans is controlled by a large family of initiating enzymes with variable expression in cells and tissues and hence is difficult to predict. Here, we used our well-established O-glycoproteomic workflows to map the precise positions of O-linked glycosylation sites on three different entities of protein S—insect cell or human cell-produced ectodomains, or insect cell derived receptor binding domain (RBD). In total 25 O-glycosites were identified, with similar patterns in the two ectodomains of different cell origin, and a distinct pattern of the monomeric RBD. Strikingly, 16 out of 25 O-glycosites were located within three amino acids from known N-glycosites. However, O-glycosylation was primarily found on peptides that were unoccupied by N-glycans, and otherwise had low overall occupancy. This suggests possible complementary functions of O-glycans in immune shielding and negligible effects of O-glycosylation on subunit vaccine design for SARS-CoV-2. MDPI 2021-03-25 /pmc/articles/PMC8064498/ /pubmed/33806155 http://dx.doi.org/10.3390/v13040551 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ). |
spellingShingle | Article Bagdonaite, Ieva Thompson, Andrew J. Wang, Xiaoning Søgaard, Max Fougeroux, Cyrielle Frank, Martin Diedrich, Jolene K. Yates, John R. Salanti, Ali Vakhrushev, Sergey Y. Paulson, James C. Wandall, Hans H. Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells |
title | Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells |
title_full | Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells |
title_fullStr | Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells |
title_full_unstemmed | Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells |
title_short | Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells |
title_sort | site-specific o-glycosylation analysis of sars-cov-2 spike protein produced in insect and human cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064498/ https://www.ncbi.nlm.nih.gov/pubmed/33806155 http://dx.doi.org/10.3390/v13040551 |
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