Advances in glycoscience to understand viral infection and colonization

Dugan, Amanda; Peiffer, Amanda L; Kiessling, Laura L.

doi:10.1038/s41592-022-01451-0

Cited by 16 publications

(15 citation statements)

References 40 publications

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“…We measured site-specific N-glycosylation occupancy and glycoform abundance at the acquired N20 and N188 N-glycosylation sites, and at the native N-glycosylation site N17. To increase the accuracy with which we could measure site-specific N-glycosylation occupancy, we aliquoted pools of peptides and N-glycopeptides into two equal portions, one for endoglycosidase treatments with peptide-N-Glycosidase F (PNGase F) in 18 O water to obtain de-glycosylated peptides, and one left untreated for intact glycopeptide analyses. PNGase F removes N-glycans converting the previously glycosylated Asn to Asp which results in a mass shift of +0.984 compared to unmodified Asn.…”

Section: Gammamentioning

confidence: 99%

“…PNGase F removes N-glycans converting the previously glycosylated Asn to Asp which results in a mass shift of +0.984 compared to unmodified Asn. In the presence of 18 O water, the mass difference is increased to +2.984 Da which increases the confidence of site assignment, particularly when multiple N-glycosylation sites are potentially present on the same peptide. Using LC-MS/MS, we then measured the abundance of the resulting peptides and de-glycosylated peptides (Supplementary Data 1) and validated N-glycosylation occupancy through the identification of glycopeptides (Supplementary Data 2).…”

Section: Gammamentioning

confidence: 99%

“…The results from the Sequest HT search were investigated using a modified version of an in-house Python script 63 . Each Asn residue in an N-glycosylation consensus site from identified PSMs (with AUC values greater than 1x10^6) was assigned U (unmodified), D (deamidated), 18 O-D (deamidated). Occupancy was defined as the proportion of the ion intensity peak area of a peptide with U or D (unmodified) or 18 O-D (occupied) identified in all charge states to the sum of the intensities of all peptide ions.…”

Section: Data Analysis N-glycosylation Occupancy At N17 N20 and N188mentioning

confidence: 99%

“…This peptide motif binds with high affinity to the active site of the oligosaccharyltransferase, the enzyme that catalyses N-glycosylation in the ER 15 . Glycans play critical roles in virus-host interactions, including stabilising the conformation of viral and host proteins; serving as viral attachment factors, co-receptors, or receptors; promoting structural conformations that facilitate or enhance receptor binding; shielding or presenting underlying viral epitopes; and acting as antigens to direct immune respones [16][17][18] . Indeed, the spike protein is heavily glycosylated, with 22 occupied N-glycosylation sequons present per protomer 19 and glycosylation has been shown to influence SARS-CoV-2 infection 20 .…”

Section: Introductionmentioning

confidence: 99%

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The role ofN-glycosylation in spike antigenicity for the SARS-CoV-2 Gamma variant

Pegg

Modhiran

Parry

et al. 2023

Preprint

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The emergence of SARS-CoV-2 variants alters the efficacy of existing immunity towards the viral spike protein, whether acquired from infection or vaccination. Mutations that impactN-glycosylation of spike may be particularly important in influencing antigenicity, but their consequences are difficult to predict. Here, we compare the glycosylation profiles and antigenicity of recombinant viral spike of ancestral Wu-1 and the Gamma strain, which has two additionalN-glycosylation sites due to amino acid substitutions in the N-terminal domain (NTD). We found that a mutation at residue 20 from threonine to asparagine within the NTD caused the loss of NTD-specific antibody binding. Glycan site-occupancy analyses revealed that the mutation resulted inN-glycosylation switching to the new sequon at N20 from the native N17 site. Site-specific glycosylation profiles demonstrated distinct glycoform differences between Wu-1, Gamma, and selected NTD variant spike proteins, but these did not affect antibody binding. Finally, we evaluated the specificity of spike proteins against convalescent COVID-19 sera and found reduced cross-reactivity against some mutants, but not Gamma spike compared to Wuhan spike. Our results illustrate the impact of viral divergence on spike glycosylation and SARS-CoV-2 antibody binding profiles.

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Section: Gammamentioning

confidence: 99%

Section: Gammamentioning

confidence: 99%

Section: Data Analysis N-glycosylation Occupancy At N17 N20 and N188mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The role ofN-glycosylation in spike antigenicity for the SARS-CoV-2 Gamma variant

Pegg

Modhiran

Parry

et al. 2023

Preprint

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“…Viral genome sequencing readily reveals amino acid mutations in emerging viral strains, yet alterations to protein glycosylation cannot be predicted from sequencing information alone. New tools to rapidly profile changes in viral protein glycosylation and their impact on immune recognition would be valuable assets in monitoring viral pathogens …”

Section: Introductionmentioning

confidence: 99%

Lectin Fingerprinting Distinguishes Antibody Neutralization in SARS-CoV-2

et al. 2023

Self Cite

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Enveloped viruses co-opt host glycosylation pathways to decorate their surface proteins. As viruses evolve, emerging strains can modify their glycosylation patterns to influence host interactions and subvert immune recognition. Still, changes in viral glycosylation or their impact on antibody protection cannot be predicted from genomic sequences alone. Using the highly glycosylated SARS-CoV-2 Spike protein as a model system, we present a lectin fingerprinting method that rapidly reports on changes in variant glycosylation state, which are linked to antibody neutralization. In the presence of antibodies or convalescent and vaccinated patient sera, unique lectin fingerprints emerge that distinguish neutralizing versus non-neutralizing antibodies. This information could not be inferred from direct binding interactions between antibodies and the Spike receptor-binding domain (RBD) binding data alone. Comparative glycoproteomics of the Spike RBD of wild-type (Wuhan-Hu-1) and Delta (B.1.617.2) variants reveal O-glycosylation differences as a key determinant of immune recognition differences. These data underscore the interplay between viral glycosylation and immune recognition and reveal lectin fingerprinting to be a rapid, sensitive, and high-throughput assay to distinguish the neutralization potential of antibodies that target critical viral glycoproteins.

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Artificial Intelligence Applications for Producing Glycosylated Biopharmaceutical Drug Modalities

von Horsten

2024

Management for Professionals

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Advances in glycoscience to understand viral infection and colonization

Cited by 16 publications

References 40 publications

The role ofN-glycosylation in spike antigenicity for the SARS-CoV-2 Gamma variant

The role ofN-glycosylation in spike antigenicity for the SARS-CoV-2 Gamma variant

Lectin Fingerprinting Distinguishes Antibody Neutralization in SARS-CoV-2

Artificial Intelligence Applications for Producing Glycosylated Biopharmaceutical Drug Modalities

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