Strategies for Proteome-Wide Quantification of Glycosylation Macro- and Micro-Heterogeneity

Abstract

Protein glycosylation governs key physiological and pathological processes in human cells. Aberrant glycosylation is thus closely associated with disease progression. Mass spectrometry (MS)-based glycoproteomics has emerged as an indispensable tool for investigating glycosylation changes in biological samples with high sensitivity. Following rapid improvements in methodologies for reliable intact glycopeptide identification, site-specific quantification of glycopeptide macro- and micro-heterogeneity at the proteome scale has become an urgent need for exploring glycosylation regulations. Here, we summarize recent advances in N- and O-linked glycoproteomic quantification strategies and discuss their limitations. We further describe a strategy to propagate MS data for multilayered glycopeptide quantification, enabling a more comprehensive examination of global and site-specific glycosylation changes. Altogether, we show how quantitative glycoproteomics methods explore glycosylation regulation in human diseases and promote the discovery of biomarkers and therapeutic targets.

The role of N-glycosylation modification in the pathogenesis of liver cancer

“…Therefore, targeting N-glycosylation is a promising approach for cancer therapy. Cancer stages have been demonstrated to be closely related to the structural specificity of N-glycosylation, and identifying certain glycosylated epitopes in tumor tissue will aid in patient staging [ 122 ]. In addition, much evidence has supported the feasibility of targeting N-glycosylation in immunotherapy [ 123 ].…”

Proteomics and glycoproteomics of beer and wine

“…Specialised software is required identify the glycan component attached to peptides or proteins, and MS2 spectra from these species are more complex owing to glycan fragmentation. In addition, glycosylated species are typically lower in abundance than peptides due to glycan heterogeneity and reduced ionisation efficacy during MS. For MS‐based quantification of glycosylation events, label and label‐free approaches including DDA, DIA and multiple reaction monitoring (MRM) are commonly used [101].…”

“…For MS-based quantification of glycosylation events, label and label-free approaches including DDA, DIA and multiple reaction monitoring (MRM) are commonly used [101].…”

Self-assembly of hydrazide-linked porous organic polymers rich in titanium for efficient enrichment of glycopeptides and phosphopeptides from human serum

“…[3][4][5] It has been discovered that several human diseases, such as cancers and neurodegenerative diseases, are intimately related to abnormal glycosylation and phosphorylation. [6][7][8] Until now, research work has concentrated on the individual functions of glycosylation or phosphorylation in cellular activity. However, these two PTMs usually interact with each other and do not act on a particular biologi-cal process separately.…”