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 carri...

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Veröffentlicht in:	Analytical chemistry (Washington) 2013-09, Vol.85 (18), p.8475-8482
Hauptverfasser:	Ohtake, Atsuko, Daikoku, Shusaku, Suzuki, Katsuhiko, Ito, Yukishige, Kanie, Osamu
Format:	Artikel
Sprache:	eng
Schlagworte:	Analytical chemistry Animals Biological Cells Cellular Cercopithecus aethiops Chromatography Chromatography, Liquid - methods COS Cells Cytoplasm - chemistry Enzymes Fluorescence Fluorescent Dyes - chemistry Glycan Glycosphingolipids - analysis Lipids Mass spectrometry Nanostructure Nanotechnology - methods Spectrometry, Mass, Electrospray Ionization - methods Tandem Mass Spectrometry - methods Transformations
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Zusammenfassung:	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.
ISSN:	0003-2700 1520-6882
DOI:	10.1021/ac401632t