Citation:
BAO Huimin, XIE Liqi, LU Haojie. Research advances of enrichment approaches based on chemical reactions in glycoproteomics[J]. Chinese Journal of Chromatography,
;2016, 34(12): 1145-1153.
doi:
10.3724/SP.J.1123.2016.09003
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Protein glycosylation is one of the most common and important post-translational modifications (PTMs) in mammalian cells. Glycopeptides usually exist at relatively low abundance (less than 5%) compared with non-glycosylated peptides. So, enrichment approaches are important for the analysis of glycoproteome. Approaches based on chemical reactions are the main trend, here we discuss them from chemical viewpoint. There are two main processes for enrichment of glycoproteins, glycoprotein conjugation and the release from solid phase. In conjugation process, we introduce four types of chemical reactions:one is boronic acid chemistry based on reaction between boronic acid and cis-diol groups on free glycans, and the other three are covalent reactions between aldehyde groups and functionalized groups, hydrazide for hydrazone ligation, alkylamine for reductive amination, aniline for nonreductive amination and aminooxy group for oxime click chemistry. In release process, N-glycosidase (PNGase) F for N-glycosylated proteins, β-elimination reaction for O-glycosylated proteins and a bleach method published recently are introduced, and the oxidative releasing of natural glycans is also introduced here. In addition, analysis strategies for N-/O-glycoproteins are discussed, including detection in glycoproteins and site-specific glycan occupancies. This review mainly focuses on the currently available chemical approaches based on covalent reactions for enrichment of glycoproteins, as well as new materials derivatized with different functional groups. We compare the advantages and disadvantages of different methods and try to evaluate the contribution and future trend based on authors viewpoint.
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