Mass spectrometric analysis of sulfated N- and O-glycans.
Sulfated N- and O-glycans carried on a myriad of cell-surface adhesion molecules and receptors are often not detected by current approaches in mass spectrometry (MS)-based glycomic mapping of cells and tissues. This is in part due to a natural lower abundance, compounded further by their negatively charged nature, which adversely disfavors their ionization and detection amid a sea of often much more abundant, nonsulfated, sialylated glycans. However, this particular limitation can actually be taken advantage of to effect highly selective enrichment and sensitive MS screening in negative ion mode, provided the ubiquitous sialic acids can first be neutralized. It has been demonstrated that permethylation not only fulfills this role adequately but further confers better MS/MS fragmentation characteristics for more efficient structural mapping and sequencing. Protocols and general practical considerations are described here which would enable one to readily prepare permethylated sulfated glycans, fractionate them away from the more abundant nonsulfated ones in simple steps for high-sensitivity MS analysis, and sensibly interpret the initial sulfoglycomic screening data thus obtained.
Khoo
Kay-Hooi
KH
Institute of Biological Chemistry, Academia Sinica, Nankang, Taipei, Taiwan.
Yu
Shin-Yi
SY
D013058:Mass Spectrometry / Q000379:methods*; D009584:Nitrogen / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry / Q000145:classification
D011134:Polysaccharides; D009584:Nitrogen; D010100:Oxygen