Analysis of the Cellular Dynamics of Fluorescently Tagged Glycosphingolipids by Using a Nanoliquid Chromatography-Tandem Mass Spectrometry Platform

Despite the increasing biological interests on glycoconjugates, the synthetic mechanism of oligosaccharides has not yet been revealed except for the enzymes involved. To clarify the synthetic events that occur inside cells, spatiotemporal analysis of fluorescently tagged glycosphingolipids was carried out. Transformation of the incorporated lactosylceramide analogue carrying 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl group (BODIPY fluorophore) was analyzed using nanoLC-fluorescence detection-nanoelectrospray ionization-mass spectrometry. A complex process of glycan synthesis and degradation was our primary observation.