Novel Technologies for Quantitative O-Glycomics and Amplification/Preparation of Cellular O-Glycans

Mucin-type O-glycosylation (O-glycans, O-glycome) characterized by GalNAc linked to Serine/Threonine or even tyrosine residues in proteins is one of the major types of glycosylations. In animals, O-glycans on glycoproteins participate in many critical biological processes such as cell adhesion, development, and immunity. Importantly, the O-glycome is different in a tissue/cell-specific manner, and often altered in cells at their pathological states; and this alteration, in turn, affects cellular properties and functions. Clearly, the Functional O-glycomics, which concerns biological roles of O-glycans, requires a comprehensive understanding of O-glycome. Structural and/or quantitative analysis of O-glycans, however, is an unmet demand because no enzyme can universally release O-glycans from glycoproteins. Furthermore, the preparation of complex O-glycans for biological studies is even more challenging. To meet these demands, we have developed a novel technology termed Cellular O-glycome Reporter/Amplification (CORA) for profiling cellular O-glycan structures and amplifying/preparing complex O-glycans from cultured cells. In this chapter, we describe the recent advances of CORA: quantitative-CORA (qCORA) and preparative-CORA (pCORA). qCORA takes the strategy of “metabolic stable isotopic labeling O-glycome of culture cells (SILOC),” and pCORA adapts cells to “O-glycan factories” when supplied with R-α-GalNAc(Ac)3 derivatives. qCORA and pCORA technologies can facilitate the cellular O-glycomics and functional O-glycomics studies.