Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins

Yong, Zhang; Zhao, Wanjun; Mao, Yonghong; Chen, Yaohui; Wang, Shisheng; Zhong, Yi; Su, Tan; Gong, Meng; Du, Dan; Lu, Xiaofeng; Cheng, Jingqiu; Yang, Hao

doi:10.1074/mcp.ra120.002295

Cited by 120 publications

(129 citation statements)

References 48 publications

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“…When the site-specific N-linked glycans are mapped onto the prefusion structure of the SARS-CoV-2 S ectodomain ( 63 ), the resulting model exhibited substantially higher levels of glycan-free surface than that revealed by structures of fully glycosylated, trimeric HIV-1 Env ectodomains ( 65 , 66 ). This suggests that the SARS-CoV-2 S protein is covered by a less dense and less effective glycan shield compared to viral glycoproteins from HIV-1 ( 36 , 66 ) and Lassa virus ( 67 ), which may be beneficial for the induction of humoral immunity and could be good news for a SARS-CoV-2 vaccine ( 68 ).…”

Section: Glycan Shield Of the Sars-cov-2 S Glycoproteinmentioning

confidence: 99%

The SARS-CoV-2 Spike Glycoprotein Biosynthesis, Structure, Function, and Antigenicity: Implications for the Design of Spike-Based Vaccine Immunogens

et al. 2020

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The ongoing pandemic of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), poses a grave threat to global public health and imposes a severe burden on the entire human society. Like other coronaviruses, the SARS-CoV-2 genome encodes spike (S) glycoproteins, which protrude from the surface of mature virions. The S glycoprotein plays essential roles in virus attachment, fusion and entry into the host cell. Surface location of the S glycoprotein renders it a direct target for host immune responses, making it the main target of neutralizing antibodies. In the light of its crucial roles in viral infection and adaptive immunity, the S protein is the focus of most vaccine strategies as well as therapeutic interventions. In this review, we highlight and describe the recent progress that has been made in the biosynthesis, structure, function, and antigenicity of the SARS-CoV-2 S glycoprotein, aiming to provide valuable insights into the design and development of the S protein-based vaccines as well as therapeutics.

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Section: Glycan Shield Of the Sars-cov-2 S Glycoproteinmentioning

confidence: 99%

The SARS-CoV-2 Spike Glycoprotein Biosynthesis, Structure, Function, and Antigenicity: Implications for the Design of Spike-Based Vaccine Immunogens

et al. 2020

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“…The studies also detected one O-glycopeptide occupied at sites, distinct from the predicted furin cleavage site at the S1/S2 boundary. 6 , 8 , 9 …”

Section: Introductionmentioning

confidence: 99%

N- and O-Glycosylation of the SARS-CoV-2 Spike Protein

2021

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Covid-19 pandemic outbreak is the reason of the current world health crisis. The development of effective antiviral compounds and vaccines requires detailed descriptive studies of SARS-CoV-2 proteins. The SARS-CoV-2 spike (S) protein mediates virion binding to the human cells through its interaction with the ACE2 cell surface receptor and is one of the prime immunization targets. A functional virion is composed of three S1 and three S2 subunits created by furin cleavage of the spike protein at R682, a polybasic cleavage site that differs from the SARS-CoV spike protein of 2002. By analysis of the protein produced in HEK293 cells, we observe that the spike is O-glycosylated on a threonine (T678) near the furin cleavage site occupied by core-1 and core-2 structures. In addition, we have identified eight additional O-glycopeptides on the spike glycoprotein and confirmed that the spike protein is heavily N-glycosylated. Our recently developed liquid chromatography−mass spectrometry methodology allowed us to identify LacdiNAc structural motifs on all occupied N-glycopeptides and polyLacNAc structures on six glycopeptides of the spike protein. In conclusion, our study substantially expands the current knowledge of the spike protein's glycosylation and enables the investigation of the influence of O-glycosylation on its proteolytic activation.

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“…The exact structures of at least 22 N-linked glycans were successfully established by mass spectrometry [17] and independently confirmed. [18] In addition, the nucleocapsid protein is decorated with O-glycans and N-glycans, at 7 and 2 confirmed sites, respectively. [19] The glycans of the spike protein act as a shield to thwart the host immune response, but a recent study has shown that N-glycans at two sites (N165 & N234) modulate the conformational dynamics of the receptor-binding domain (RBD).…”

Section: The Sars-cov-2 Machinery Landscapementioning

confidence: 99%

Proteomics in the COVID‐19 Battlefield: First Semester Check‐Up

Grenga

Armengaud

2020

Proteomics

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Proteomics offers a wide collection of methodologies to study biological systems at the finest granularity. Faced with COVID‐19, the most worrying pandemic in a century, proteomics researchers have made significant progress in understanding how the causative virus hijacks the host's cellular machinery and multiplies exponentially, how the disease can be diagnosed, and how it develops, as well as its severity predicted. Numerous cellular targets of potential interest for the development of new antiviral drugs have been documented. Here, the most striking results obtained in the proteomics field over this first semester of the pandemic are presented. The molecular machinery of SARS‐CoV‐2 is much more complex than initially believed, as many post‐translational modifications can occur, leading to a myriad of proteoforms and a broad heterogeneity of viral particles. The interplay of protein–protein interactions, protein abundances, and post‐translational modifications has yet to be fully documented to provide a full picture of this intriguing but lethal biological threat. Proteomics has the potential to provide rapid detection of the SARS‐CoV‐2 virus by mass spectrometry proteotyping, and to further increase the knowledge of severe respiratory syndrome COVID‐19 and its long‐term health consequences.

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Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins

Cited by 120 publications

References 48 publications

The SARS-CoV-2 Spike Glycoprotein Biosynthesis, Structure, Function, and Antigenicity: Implications for the Design of Spike-Based Vaccine Immunogens

The SARS-CoV-2 Spike Glycoprotein Biosynthesis, Structure, Function, and Antigenicity: Implications for the Design of Spike-Based Vaccine Immunogens

N- and O-Glycosylation of the SARS-CoV-2 Spike Protein

Proteomics in the COVID‐19 Battlefield: First Semester Check‐Up

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