Identification and Characterization of N-Glycosylated Proteins Using Proteomics

Glycoproteins constitute a large fraction of the proteome. The fundamental role of protein glycosylation in cellular development, growth, and differentiation, tissue development, and in host–pathogen interactions is by now widely accepted. Proteome–wide characterization of glycoproteins is a complex task and is currently achieved by mass spectrometry–based methods that enable identification of glycoproteins and localization, classification, and analysis of individual glycan structures on proteins. In this chapter we briefly introduce a range of analytical technologies for recovery and analysis of glycoproteins and glycopeptides. Combinations of affinity–enrichment techniques, chemical and biochemical protocols, and advanced mass spectrometry facilitate detailed glycoprotein analysis in proteomics, from fundamental biological studies to biomarker discovery in biomedicine.