Tomohisa Kato

Development of tetraphenylethylene-based fluorescent oligosaccharide probes for detection of influenza virus.

Abstract Tetraphenylethylene (TPE) derivatives have strong fluorescence in aggregated state. We designed and synthesized a tetraphenylethylene derivative bearing alkyne groups which were used for combination by click chemistry. The new TPE compound bearing alkyne groups was used to synthesize fluorescence oligosaccharide probes which have lactosyl and 6′-sialyllactosyl moieties as ligands. We found that the TPE compounds bearing lactosyl and 6′-sialyllactosyl moieties were useful for detection of RCA120 and SSA lectins, respectively. Moreover, we have shown that TPE-based fluorescent oligosaccharide probe bearing 6′-sialyllactose moiety can be utilized as a “turn-on” fluorescent sensor for detection of influenza virus.

Novel method for chase analysis of oligosaccharide metabolic error caused by xenobiotics.

Saccharide primers, such as dodecyl beta-lactoside (Lac-C12), are unique artificial precursors of glycolipid synthesis. In culture media supplemented with saccharide primers, they are taken up by the cells in the culture media and glycosylated by cellular glycosyltransferases in the Golgi apparatus to form pseudo-glycolipids. In this study, we examine the effects of various xenobiotics on glycolipid synthesis by implementing a novel method to analyze pseudo-glycolipids, mainly gangliosides, produced by ONS-76 medulloblastoma cells in a culture medium containing various xenobiotics. The ganglioside group of pseudo-glycolipids was effectively purified by using strong anion-exchange cartridges. The production of pseudo-gangliosides was stimulated by N-(n-butyl)deoxygalactonojirimycin (NB-DGJ), but was inhibited by castanospermine, 2-deoxy-2-fluoro-d-galactose, tunicamycin, and brefeldin A. Because the cells in the culture medium are exposed to the saccharide primers and xenobiotics at the same time, and are secreted in the culture medium in their glycosylated form, our method can be used to effectively analyze the direct effects of xenobiotics on ganglioside synthesis.