O-Glycoproteomics of Viruses
Global O-glycoproteomics of viruses offers the possibility of rapidly “scanning” the proteome of viruses for O-glycan modifications. The strategy can be applied to multiple viruses that their relevant propagation systems are available. Based on the advanced equipment and experienced scientists, Creative Proteomics is capable of offering O-glycoproteomics services. Interfacing the mass spectrometer with separation technologies (such as liquid chromatography and capillary electrophoresis), we are able to separate complex proteome-scale glycopeptide mixtures and identify thousands of glycopeptides in a single run. Our services can provide the formation of structure heterogeneity, exact position, and site occupancy for each individual glycosite at a single protein level.
O-linked glycosylation of viral proteins
O-glycosylation is found on the side chains of the hydroxylamino acids Serine and Threonine (the “O” indicating attachment of sugar residues through the side chain hydroxyl group). Unlike N-glycosylation, O-glycosylation is not controlled by the presence of a linear consensus sequence in the amino acid chain, thus O-glycosylation prediction is not possible from the linear amino acid sequence. In the past decades, the study of site-specific O-glycosylation of viral proteins has been limited. Thanks to the advances in mass spectrometry-based proteomics, there is more and more information available regarding the O-glycosylation of individual viral glycoproteins. So far, O-glycoproteomic analyses of several recombinant or isolated viral glycoproteins have been investigated, including influenza A virus HA1, HIV-1 gp120, HSV-1 gC, HCV E2, and Hendra virus glycoprotein G.
Service offering in Creative Proteomics
The enrichment of glycopeptides is a critical step, especially for sensitive glycoproteomics. Based on peanut agglutinin (PNA) lectin weak affinity chromatography (LWAC) and Vicia villosa lectin (VVA) LWAC, we established a two-step lectin enrichment strategy to identify O-glycosites. In addition, the structures of O-glycans are characterized by high heterogeneity, which is associated with both cell type and protein specific, thereby, we carefully select candidates and expression cell lines for specific research and applications. Unlike N-glycosylation, there is a particular need to perform experimental identification of O-glycosites. The season is that O-glycosites lack simple predictive consensus sequence motifs and the o-glycosylation capacity of host cells must also be considered. Finally, our services are able to comprehensively characterize O-glycoproteomes of any virus produced in infected host cells.
The workflow of our services
Advantages of our services
- The glycoproteomic strategy employed is highly sensitive
- Allow to analyze all viral glycoproteins expressed in host cells simultaneously
- Involve endogenous glycosylation of the cells, which is regulated by glycosyltransferases
- Take into account the cytopathic effects of viral infection as well as native conformations of proteins
O-linked glycosylation may take part in viral entry, formation, secretion, and immune recognition, and the O-glycoproteomics services presented here are now open for unbiased discovery on all enveloped viruses. Creative Proteomics is committed to providing high-quality service to greatly accelerate our customers' project progress and success. For more information on how we can help you, please feel free to contact us. Customers have direct access to our staff and get timely feedback.
Reference
- Cipollo, J. F., & Parsons, L. M. (2020). “Glycomics and glycoproteomics of viruses: Mass spectrometry applications and insights toward structure–function relationships.” Mass Spectrometry Reviews, 39(4), 371-409.
* For research use only.
