O-Linked Glycoproteomics Sample Preparation
There are many types of protein glycosylation in viral research, and N- and mucin-type O-linked glycosylation are the most studied. Since the machinery of N-linked and O-linked glycosylation is different, specific types of sample preparation and analyses are required for each of these glycosylation types. Based on a team of professional and experienced scientists, Creative Proteomics is capable of offering O-linked glycoproteomics sample preparation services for mass spectrometric study. We are committed to providing high-quality service to greatly accelerate our customers' project progress and success.
O-linked glycosylation
Protein glycosylation is the most diverse and complex form of protein modification, which greatly improves protein heterogeneity and promotes functional plasticity. Compared to N-linked glycosylation, the characterization of O-linked glycosylation is even more challenging due to the complex structure of the added polysaccharides as well as the technical challenges of definitively characterizing them. O-linked glycosylation is mainly located in the Golgi apparatus and can respond to the changes of various internal and external factors. Although the majority of O-glycosylation occurs through the secretory pathway, cytosolic and nuclear proteins can also be glycosylated with the transfer of N-acetylglucosamine (GlcNAc) via a cytosolic glycosyltransferase. Therefore, O-linked glycosylation shows high heterogeneity in different cells, tissues, and diseases.
O-linked glycoproteomics sample preparation
Based on professional knowledge and experienced experts, we are proud to offer novel workflows for efficient O-glycopeptide generation for virus-infected cells. Our services mainly include the removal of N-linked glycans, the enrichment of O-linked glycopeptides, and the effective digestion of O-glycoproteins.
The removal of N-linked glycans
This is a key step prior to subsequent LC-MS/MS analysis, which reduces the complexity of the sample, improves the digestion, as well as facilitates the correct assignments of O-glycosite.
The enrichment of O-linked glycopeptides
The enrichment of O-linked glycopeptides from complex biological samples is essential for the identification of O-linked glycoproteome, especially for the low abundance O-linked glycoproteome. To achieve the goal, a series of enrichment methodologies have been applied to study O-linked glycoproteome in different biological systems, such as lectins (PNA and VVA lectins), hydrazide chemistry, metabolic labeling, and hydrop interaction liquid chromatography (HILIC).
The effective digestion of O-glycoproteins
The digestion of O-glycoproteins can be effectively achieved by using novel glycoproteases, such as glycoproteases that specifically cleave C-terminally (StcE) or N-terminally (OpeRATOR) from O-glycosylated serines and threonines.
Benefits of our services
- Professional technical support
- Rapid and efficient service, allowing investigation of large sample cohorts
- Highly reproducible sample preparation
- Customized service to customer satisfaction
Our services can be implemented for the proteome-wide O-glycan characterization of multiple viruses. For any requested information for our O-linked glycoproteomic sample preparation services, please contact us. We will reply to your message soon!
How to order?
Reference
- Yang, W., et al. (2018). "Mapping the O‐glycoproteome using site‐specific extraction of O‐linked glycopeptides (EXoO)." Molecular systems biology, 14(11), e8486.
* For research use only.
