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

Breimer, Michael E.; Hansson, Gunnar C.; Karlsson, Karl-Anders; Larson, Goran and Leffler, Hakon LU (2012) In Glycobiology 22(12). p.1721-1730
Abstract
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 ethylenediaminetetraacetic acid-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, Escherichia coli) detection reagents, mass spectrometry and proton NMR spectroscopy of derivatized glycolipids. In this way, the structure of major... (More)
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 ethylenediaminetetraacetic acid-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, Escherichia 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 the dominance of monoglycosylceramides, sulphatides and blood group fucolipids (mainly Le(b) hexaglycosylceramides and ALe(b) heptaglycosylceramides). The non-epithelial residue had about five 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 with the 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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cholera toxin receptor, epithelial cells, glycosphingolipids, human, small intestine, lipid rafts, Norovirus receptor
in
Glycobiology
volume
22
issue
12
pages
1721 - 1730
publisher
Oxford University Press
external identifiers
  • wos:000310630900007
  • scopus:84868557924
ISSN
1460-2423
DOI
10.1093/glycob/cws115
language
English
LU publication?
yes
id
e62de43c-1c0d-4dfd-9644-a376afd5812f (old id 3243079)
date added to LUP
2013-01-07 09:32:58
date last changed
2017-10-01 04:12:11
@article{e62de43c-1c0d-4dfd-9644-a376afd5812f,
  abstract     = {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 ethylenediaminetetraacetic acid-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, Escherichia 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 the dominance of monoglycosylceramides, sulphatides and blood group fucolipids (mainly Le(b) hexaglycosylceramides and ALe(b) heptaglycosylceramides). The non-epithelial residue had about five 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 with the 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.},
  author       = {Breimer, Michael E. and Hansson, Gunnar C. and Karlsson, Karl-Anders and Larson, Goran and Leffler, Hakon},
  issn         = {1460-2423},
  keyword      = {cholera toxin receptor,epithelial cells,glycosphingolipids,human,small intestine,lipid rafts,Norovirus receptor},
  language     = {eng},
  number       = {12},
  pages        = {1721--1730},
  publisher    = {Oxford University Press},
  series       = {Glycobiology},
  title        = {Glycosphingolipid composition of epithelial cells isolated along the villus axis of small intestine of a single human individual},
  url          = {http://dx.doi.org/10.1093/glycob/cws115},
  volume       = {22},
  year         = {2012},
}