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bioRxiv · 10.1101/803494

A Chemoenzymatic Method for Glycoproteomic N-glycan Type Quantitation

Li, H.·Li, L.·Cheng, K.·Ning, Z.·Mayne, J.·Zhang, X.·Walker, K.·Chen, R.·Twine, S.·Li, J.·Figeys, D.
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Abstract

Glycosylation is one of the most important post-translational modifications in biological systems. Current glycoproteome methods mainly focus on qualitative identification of glycosylation sites or intact glycopeptides. However, the systematic quantitation of glycoproteins has remained largely unexplored. Here, we developed a chemoenzymatic method to quantitatively investigate N-glycoproteome based on the N-glycan types. Taking advantage of the specificity of different endoglycosidases and isotope dimethyl labeling, six N-glycan types of structures linked on each glycopeptide, including high-mannose/hybrid, bi-antennary and tri-antennary with/without core fucose, were quantified. As a proof of principle, the glycoproteomic N-glycan type quantitative (glyco-TQ) method was first used to determine the N-glycan type composition of immunoglobulin G1 (IgG1) Fc fragment. Then we applied the method to analyze the glycan type profile of proteins in the breast cancer cell line MCF7, and quantitatively revealed the N-glycan type micro-heterogeneity at both the glycopeptide and glycoprotein levels. The novel quantitative strategy to evaluate the relative intensity of the six states of N-glycan type glycosylation on each site provides a new avenue to investigate function of glycoproteins in broad areas, such as cancer biomarker research, pharmaceuticals characterization and anti-glycan vaccine development.

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Li, H., Li, L., Cheng, K., Ning, Z., Mayne, J., Zhang, X., Walker, K., Chen, R., Twine, S., Li, J., Figeys, D.. 2019-10-15. A Chemoenzymatic Method for Glycoproteomic N-glycan Type Quantitation. https://doi.org/10.1101/803494

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