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On-line nano-HPLC/ESI QTOF MS monitoring of α2–3 and α2–6 sialylation in granulocyte glycosphingolipidome
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Stephan Kirsch
Published/Copyright:
April 10, 2009
Abstract
A novel glycosphingolipidomic protocol using nano-high performance liquid chromatography coupled on-line to electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-QTOF-MS) focusing on the separation of isomeric ganglioside structures is described here. A highly efficient separation of α2–3- and α2–6-sialylated ganglioside species of different carbohydrate chain length was achieved on an HILIC-amido column, followed by sensitive flow-through ESI-QTOF-MS detection and unambiguous structural identification by tandem MS experiments. The protocol was applied to encompass the glycosphingolipidome of human granulocytes, where 182 distinct components could be clearly identified and assigned regarding the ganglioside type and the isomer distribution.
Keywords: gangliosides and neolacto-series; glycosphingolipids; human granulocytes; nano-HPLC/MS; sialic acid linkage isomers
Received: 2009-1-21
Accepted: 2009-3-22
Published Online: 2009-04-10
Published in Print: 2009-07-01
©2009 by Walter de Gruyter Berlin New York
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Keywords for this article
gangliosides and neolacto-series;
glycosphingolipids;
human granulocytes;
nano-HPLC/MS;
sialic acid linkage isomers
Articles in the same Issue
- Guest Editorial
- Highlight: Perspectives in glycobiology
- Cell biology and glycosylation: protein targeting by O- and N-linked glycosylation
- Glycosylation- and phosphorylation-dependent intracellular transport of lysosomal hydrolases
- Glycosylation pattern of brush border-associated glycoproteins in enterocyte-like cells: involvement of complex-type N-glycans in apical trafficking
- Impact of glycosylation and detergent-resistant membranes on the function of intestinal sucrase-isomaltase
- MUC1 traverses apical recycling endosomes along the biosynthetic pathway in polarized MDCK cells
- Cell biology and glycosylation: carbohydrate-mediated recognition and signaling in cell proliferation and differentiation
- From structural to functional glycomics: core substitutions as molecular switches for shape and lectin affinity of N-glycans
- Brain development needs sugar: the role of polysialic acid in controlling NCAM functions
- Beyond glycosylation: sialic acid precursors act as signaling molecules and are involved in cellular control of differentiation of PC12 cells
- Glycosylation and disease
- Management of the human mucosal defensive barrier: evidence for glycan legislation
- Regulation and pathophysiological implications of UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE) as the key enzyme of sialic acid biosynthesis
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- O-glycosylation pattern of CD24 from mouse brain
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- On-line nano-HPLC/ESI QTOF MS monitoring of α2–3 and α2–6 sialylation in granulocyte glycosphingolipidome
