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The multidimensional HPLC mapping method developed by Dr. Noriko Takahashi and coworkers has been widely recognized as a useful tool for the structural analysis of neutral and sialyl N-glycans. The utility of this method would be further extended if it could be used to analyze anionic oligosaccharides, including sulfated or glucuronylated glycans, since they play important roles in cell-cell communications. Hence, we developed HPLC maps of sulfated and glucuronylated oligosaccharides produced by recombinant sulfotransferases and glycosyltransferases. Here, we describe the HPLC map of anionic glycans and demonstrate its utility by applying this map for the characterization of sulfotransferase N-acetylglucosamine 6-O-sulfotransferase-1 (GlcNAc6ST-1) and glycosylation proˆling of in‰uenza virus. A. Introduction Glycosylation is one of the major posttranslational modiˆcations, as is evident from the fact that more than half the number of proteins are predicted to be modiˆed with glycans (1). Glycans increase the solubility and stability of proteins as well as increase the structural integrity of protein functional sites and mediate cell-cell communications and viral infections. Particularly, sulfated glycans expressed on glycoproteins play important roles in biological events such as lymphocyte homing (2), degradation of pituitary gland hormones (3), and adhesion of neural cells

Multidimensional HPLC mapping method for the structural analysis of anionic N-glycans