A Chemoenzymatic Method for Glycoproteomic N-glycan Type Quantitation
A Chemoenzymatic Method for Glycoproteomic N-glycan Type Quantitation
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.
Cheng Kai、Ning Zhibin、Zhang Xu、Li Henghui、Walker Krystal、Li Jianjun、Mayne Janice、Figeys Daniel、Li Leyuan、Twine Susan、Chen Rui
SIMM-University of Ottawa Joint Research Center in Systems and Personalized Pharmacology and Ottawa Institute of Systems Biology and Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of OttawaSIMM-University of Ottawa Joint Research Center in Systems and Personalized Pharmacology and Ottawa Institute of Systems Biology and Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of OttawaSIMM-University of Ottawa Joint Research Center in Systems and Personalized Pharmacology and Ottawa Institute of Systems Biology and Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of OttawaSIMM-University of Ottawa Joint Research Center in Systems and Personalized Pharmacology and Ottawa Institute of Systems Biology and Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of OttawaSIMM-University of Ottawa Joint Research Center in Systems and Personalized Pharmacology and Ottawa Institute of Systems Biology and Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of OttawaHuman Health Therapeutics Research Centre, National Research Council CanadaSIMM-University of Ottawa Joint Research Center in Systems and Personalized Pharmacology and Ottawa Institute of Systems Biology and Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of OttawaSIMM-University of Ottawa Joint Research Center in Systems and Personalized Pharmacology and Ottawa Institute of Systems Biology and Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa||Molecular Architecture of Life Program, Canadian Institute for Advanced ResearchSIMM-University of Ottawa Joint Research Center in Systems and Personalized Pharmacology and Ottawa Institute of Systems Biology and Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of OttawaHuman Health Therapeutics Research Centre, National Research Council CanadaHuman Health Therapeutics Research Centre, National Research Council Canada
生物科学研究方法、生物科学研究技术生物化学分子生物学
endoglycosidaseN-glycanglycoproteomicquantitationisotope dimethyl labeling
Cheng Kai,Ning Zhibin,Zhang Xu,Li Henghui,Walker Krystal,Li Jianjun,Mayne Janice,Figeys Daniel,Li Leyuan,Twine Susan,Chen Rui.A Chemoenzymatic Method for Glycoproteomic N-glycan Type Quantitation[EB/OL].(2025-03-28)[2025-04-30].https://www.biorxiv.org/content/10.1101/803494.点此复制
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