Abstract:Severe acute respiratory syndrome coronavirus 2 is the causative pathogen of the
COVID-19 pandemic which as of March 29, 2021, has claimed 2 776 175 lives
worldwide. Vaccine development efforts focus on the viral trimeric spike glycoprotein as
the main target of the humoral immune response. Viral spikes carry glycans that
facilitate immune evasion by shielding specific protein epitopes from antibody
neutralization, and antigen efficacy is influenced by spike glycoprotein production in
vivo. Therefore, immunoge… Show more
“…Despite the observed differences, we note that the site-specific glycosylation of the virally derived material outlined here is consistent with previously reported glycan analysis from virally derived S protein produced in Calu-3 lung epithelial cells. 90 …”
A central tenet in
the design of vaccines is the display of native-like
antigens in the elicitation of protective immunity. The abundance
of N-linked glycans across the SARS-CoV-2 spike protein is a potential
source of heterogeneity among the many different vaccine candidates
under investigation. Here, we investigate the glycosylation of recombinant
SARS-CoV-2 spike proteins from five different laboratories and compare
them against S protein from infectious virus, cultured in Vero cells.
We find patterns that are conserved across all samples, and this can
be associated with site-specific stalling of glycan maturation that
acts as a highly sensitive reporter of protein structure. Molecular
dynamics simulations of a fully glycosylated spike support a model
of steric restrictions that shape enzymatic processing of the glycans.
These results suggest that recombinant spike-based SARS-CoV-2 immunogen
glycosylation reproducibly recapitulates signatures of viral glycosylation.
“…Despite the observed differences, we note that the site-specific glycosylation of the virally derived material outlined here is consistent with previously reported glycan analysis from virally derived S protein produced in Calu-3 lung epithelial cells. 90 …”
A central tenet in
the design of vaccines is the display of native-like
antigens in the elicitation of protective immunity. The abundance
of N-linked glycans across the SARS-CoV-2 spike protein is a potential
source of heterogeneity among the many different vaccine candidates
under investigation. Here, we investigate the glycosylation of recombinant
SARS-CoV-2 spike proteins from five different laboratories and compare
them against S protein from infectious virus, cultured in Vero cells.
We find patterns that are conserved across all samples, and this can
be associated with site-specific stalling of glycan maturation that
acts as a highly sensitive reporter of protein structure. Molecular
dynamics simulations of a fully glycosylated spike support a model
of steric restrictions that shape enzymatic processing of the glycans.
These results suggest that recombinant spike-based SARS-CoV-2 immunogen
glycosylation reproducibly recapitulates signatures of viral glycosylation.
“…While sites N331 and N343 in the RBD region showed predominantly high mannose glycans when S protein is expressed in individual subunits S1 and S2, the same sites showed complex-type glycans (with 98% fucosylation) on trimeric form of S protein expressed on HEK293 cells [ 6 , 10 ]. A recent quantitative N-glycan analysis on S protein subunit S1 isolated from SARS-CoV-2-infected Calu-3 cells via immunoaffinity purification showed high prevalence of complex-type N-glycans (79%) and 21% high mannose and/or hybrid structures [ 16 ]. The same study compared the different glycans on vaccine candidates and recombinant S protein with the wild-type virus S protein with an aim to help the vaccine design strategies and thereby enable high-quality immune response through correct immunogen presentation.…”
Section: Sars-cov-2 Spike Protein Glycosylationmentioning
confidence: 99%
“…Recent structural analysis of the SARS-CoV-2 S protein by cryo-EM shows that it is extensively glycosylated, similar to SARS-CoV-1 S protein [ 7 – 9 ]. Moreover, the site-specific glycosylation analysis of S protein by our group and other groups through mass spectrometry revealed both N- and O-glycosylation [ 10 – 16 ]. Based on recent reports, each trimeric spike presents up to 66 N-linked glycosylation sites and several O-linked glycosylation sites [ 12 , 16 ].…”
Section: Introductionmentioning
confidence: 99%
“…Moreover, the site-specific glycosylation analysis of S protein by our group and other groups through mass spectrometry revealed both N- and O-glycosylation [ 10 – 16 ]. Based on recent reports, each trimeric spike presents up to 66 N-linked glycosylation sites and several O-linked glycosylation sites [ 12 , 16 ]. Site-specific glycosylation on virus-derived, wild-type non-stabilized and recombinant stabilized spike glycoproteins was compared in a latest study [ 16 ].…”
Section: Introductionmentioning
confidence: 99%
“…Based on recent reports, each trimeric spike presents up to 66 N-linked glycosylation sites and several O-linked glycosylation sites [ 12 , 16 ]. Site-specific glycosylation on virus-derived, wild-type non-stabilized and recombinant stabilized spike glycoproteins was compared in a latest study [ 16 ].…”
Graphical abstract
The COVID-19 pandemic is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Similar to other coronaviruses, its particles are composed of four structural proteins: spike (S), envelope (E), membrane (M), and nucleocapsid (N) proteins. S, E, and M proteins are glycosylated, and the N protein is phosphorylated. The S protein is involved in the interaction with the host receptor human angiotensin-converting enzyme 2 (hACE2), which is also heavily glycosylated. Recent studies have revealed several other potential host receptors or factors that can increase or modulate the SARS-CoV-2 infection. Interestingly, most of these molecules bear carbohydrate residues. While glycans acquired by the viruses through the hijacking of the host machinery help the viruses in their infectivity, they also play roles in immune evasion or modulation. Glycans play complex roles in viral pathobiology, both on their own and in association with carrier biomolecules, such as proteins or glycosaminoglycans (GAGs). Understanding these roles in detail can help in developing suitable strategies for prevention and therapy of COVID-19. In this review, we sought to emphasize the interplay of SARS-CoV-2 glycosylated proteins and their host receptors in viral attachment, entry, replication, and infection. Moreover, the implications for future therapeutic interventions targeting these glycosylated biomolecules are also discussed in detail.
Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) is the cause of the on‐going global pandemic of coronavirus disease 2019 (COVID‐19) that continues to pose a significant threat to public health worldwide. SARS‐CoV‐2 encodes four structural proteins namely membrane, nucleocapsid, spike, and envelope proteins that play essential roles in viral entry, fusion, and attachment to the host cell. Extensively glycosylated spike protein efficiently binds to the host angiotensin‐converting enzyme 2 initiating viral entry and pathogenesis. Reverse transcriptase polymerase chain reaction on nasopharyngeal swab is the preferred method of sample collection and viral detection because it is a rapid, specific, and high‐throughput technique. Alternate strategies such as proteomics and glycoproteomics‐based mass spectrometry enable a more detailed and holistic view of the viral proteins and host–pathogen interactions and help in detection of potential disease markers. In this review, we highlight the use of mass spectrometry methods to profile the SARS‐CoV‐2 proteome from clinical nasopharyngeal swab samples. We also highlight the necessity for a comprehensive glycoproteomics mapping of SARS‐CoV‐2 from biological complex matrices to identify potential COVID‐19 markers.
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