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Glycosphingolipid composition of epithelial cells isolated along the villus axis of small intestine of a single human individual.

Artikel i vetenskaplig tidskrift
Författare Michael Breimer
Gunnar C. Hansson
Karl-Anders Karlsson
Göran Larson
Hakon Leffler
Publicerad i Glycobiology
Volym 22
Nummer/häfte 12
Sidor 1721-1730
ISSN 1460-2423
Publiceringsår 2012
Publicerad vid Institutionen för kliniska vetenskaper, Avdelningen för kirurgi
Institutionen för biomedicin, avdelningen för klinisk kemi och transfusionsmedicin
Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi
Sidor 1721-1730
Språk en
Länkar dx.doi.org/10.1093/glycob/cws115
Ämneskategorier Klinisk medicin, Kirurgi

Sammanfattning

A 6 cm fresh proximal ileum surgical specimen from a blood group A(1)Le(a-b+) secretor individual was used for stepwise isolation of epithelial cells from villus tip to crypt bottom by gentle washing with EDTA-containing buffer. Acid and non-acid sphingolipids were prepared from the epithelial cell fractions and the non-epithelial intestinal residue. Molecular information on the sphingolipid composition was obtained without further isolation of individual species by applying thin-layer chromatography using chemical and biological (monoclonal antibodies, cholera toxin, E coli) detection reagents, mass spectrometry and proton NMR spectroscopy of derivatized glycolipids. In this way the structure of major and minor saccharides, ceramide components and their relative amounts were obtained. Epithelial cells and non-epithelial residue were distinctly different in their sphingolipid composition. Sphingomyelin was the major single component in both compartments. Characteristic for epithelial cells was dominance of monoglycosylceramides, sulphatides and blood group fucolipids (mainly Le(b) hexaglycosylceramides and ALe(b) heptaglycosylceramides). The non-epithelial residue had about 5 times less glycolipids mainly mono-, di-, tri- and tetra-glycosylceramides and gangliosides, including the GM1 ganglioside. The ceramides were more hydroxylated (1-2 additional hydroxyls) in epithelial cell glycolipids compared to non-epithelial residue. Combined with a separate detailed study on the glycoproteins of the same epithelial cell preparation, this human intestinal sample is the only epithelial cell preparation where both protein- and lipid-linked saccharides are characterized in detail. Calculation of the relative distribution of different carbohydrate determinants gives structural support for a raft like distribution of sphingolipids in the outer leaflet of the epithelial membrane.

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