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O-Glycosylation Landscapes of SARS-CoV-2 Spike Proteins
The densely glycosylated spike (S) proteins that are highly exposed on the surface of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) facilitate viral attachment, entry, and membrane fusion. We have previously reported all the 22 N-glycosites and site-specific N-glycans in the S protein...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8450404/ https://www.ncbi.nlm.nih.gov/pubmed/34552909 http://dx.doi.org/10.3389/fchem.2021.689521 |
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author | Zhang, Yong Zhao, Wanjun Mao, Yonghong Chen, Yaohui Zheng, Shanshan Cao, Wei Zhu, Jingqiang Hu, Liqiang Gong, Meng Cheng, Jingqiu Yang, Hao |
author_facet | Zhang, Yong Zhao, Wanjun Mao, Yonghong Chen, Yaohui Zheng, Shanshan Cao, Wei Zhu, Jingqiang Hu, Liqiang Gong, Meng Cheng, Jingqiu Yang, Hao |
author_sort | Zhang, Yong |
collection | PubMed |
description | The densely glycosylated spike (S) proteins that are highly exposed on the surface of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) facilitate viral attachment, entry, and membrane fusion. We have previously reported all the 22 N-glycosites and site-specific N-glycans in the S protein protomer. Herein, we report the O-glycosylation landscapes of SARS-CoV-2 S proteins, which were characterized through high-resolution mass spectrometry. Following digestion with trypsin and trypsin/Glu-C, and de-N-glycosylation using PNGase F, we determined the GalNAc-type O-glycosylation pattern of S proteins, including O-glycosites and the six most common O-glycans occupying them, via Byonic identification and manual validation. Finally, 255 intact O-glycopeptides composed of 50 peptides sequences and 43 O-glycosites were discovered by higher energy collision-induced dissociation (HCD), and three O-glycosites were confidently identified by electron transfer/higher energy collision-induced dissociation (EThcD) in the insect cell-expressed S protein. Most glycosites were modified by non-sialylated O-glycans such as HexNAc(1) and HexNAc(1)Hex (1). In contrast, in the human cell-expressed S protein S1 subunit, 407 intact O-glycopeptides composed of 34 peptides sequences and 30 O-glycosites were discovered by HCD, and 11 O-glycosites were unambiguously assigned by EThcD. However, the measurement of O-glycosylation occupancy hasn’t been made. Most glycosites were modified by sialylated O-glycans such as HexNAc(1)Hex (1)NeuAc (1) and HexNAc(1)Hex (1)NeuAc (2). Our results reveal that the SARS-CoV-2 S protein is an O-glycoprotein; the O-glycosites and O-glycan compositions vary with the host cell type. These comprehensive O-glycosylation landscapes of the S protein are expected to provide novel insights into the viral binding mechanism and present a strategy for the development of vaccines and targeted drugs. |
format | Online Article Text |
id | pubmed-8450404 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-84504042021-09-21 O-Glycosylation Landscapes of SARS-CoV-2 Spike Proteins Zhang, Yong Zhao, Wanjun Mao, Yonghong Chen, Yaohui Zheng, Shanshan Cao, Wei Zhu, Jingqiang Hu, Liqiang Gong, Meng Cheng, Jingqiu Yang, Hao Front Chem Chemistry The densely glycosylated spike (S) proteins that are highly exposed on the surface of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) facilitate viral attachment, entry, and membrane fusion. We have previously reported all the 22 N-glycosites and site-specific N-glycans in the S protein protomer. Herein, we report the O-glycosylation landscapes of SARS-CoV-2 S proteins, which were characterized through high-resolution mass spectrometry. Following digestion with trypsin and trypsin/Glu-C, and de-N-glycosylation using PNGase F, we determined the GalNAc-type O-glycosylation pattern of S proteins, including O-glycosites and the six most common O-glycans occupying them, via Byonic identification and manual validation. Finally, 255 intact O-glycopeptides composed of 50 peptides sequences and 43 O-glycosites were discovered by higher energy collision-induced dissociation (HCD), and three O-glycosites were confidently identified by electron transfer/higher energy collision-induced dissociation (EThcD) in the insect cell-expressed S protein. Most glycosites were modified by non-sialylated O-glycans such as HexNAc(1) and HexNAc(1)Hex (1). In contrast, in the human cell-expressed S protein S1 subunit, 407 intact O-glycopeptides composed of 34 peptides sequences and 30 O-glycosites were discovered by HCD, and 11 O-glycosites were unambiguously assigned by EThcD. However, the measurement of O-glycosylation occupancy hasn’t been made. Most glycosites were modified by sialylated O-glycans such as HexNAc(1)Hex (1)NeuAc (1) and HexNAc(1)Hex (1)NeuAc (2). Our results reveal that the SARS-CoV-2 S protein is an O-glycoprotein; the O-glycosites and O-glycan compositions vary with the host cell type. These comprehensive O-glycosylation landscapes of the S protein are expected to provide novel insights into the viral binding mechanism and present a strategy for the development of vaccines and targeted drugs. Frontiers Media S.A. 2021-09-06 /pmc/articles/PMC8450404/ /pubmed/34552909 http://dx.doi.org/10.3389/fchem.2021.689521 Text en Copyright © 2021 Zhang, Zhao, Mao, Chen, Zheng, Cao, Zhu, Hu, Gong, Cheng and Yang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Chemistry Zhang, Yong Zhao, Wanjun Mao, Yonghong Chen, Yaohui Zheng, Shanshan Cao, Wei Zhu, Jingqiang Hu, Liqiang Gong, Meng Cheng, Jingqiu Yang, Hao O-Glycosylation Landscapes of SARS-CoV-2 Spike Proteins |
title | O-Glycosylation Landscapes of SARS-CoV-2 Spike Proteins |
title_full | O-Glycosylation Landscapes of SARS-CoV-2 Spike Proteins |
title_fullStr | O-Glycosylation Landscapes of SARS-CoV-2 Spike Proteins |
title_full_unstemmed | O-Glycosylation Landscapes of SARS-CoV-2 Spike Proteins |
title_short | O-Glycosylation Landscapes of SARS-CoV-2 Spike Proteins |
title_sort | o-glycosylation landscapes of sars-cov-2 spike proteins |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8450404/ https://www.ncbi.nlm.nih.gov/pubmed/34552909 http://dx.doi.org/10.3389/fchem.2021.689521 |
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