Glycobiology, 2000, Vol. 10, No. 6 565-575
© 2000 Oxford University Press
Massive in vitro synthesis of tagged oligosaccharides in 1-benzyl-2-acetamido-2-deoxy-
-D-galactopyranoside treated HT-29 cells
Unité de Glycobiologie structurale et Fonctionnelle, UMR CNRS no. 8576, Laboratoire de Chimie Biologique, Université des Sciences et Technologies de Lille, F-59655 Villeneuve d’Ascq, France, 2Unité de Biologie et Physiopathologie des Cellules Mucipares, INSERM U-377, place de Verdun, F-59045 Lille, France, and 3Institut Biomédical des Cordeliers, INSERM U-505, 15 rue de l’Ecole de Médecine, F-75006 Paris, France
Permanent exposure of differentiated HT-29 cells to the sugar analogue, 1-benzyl-2-acetamido-2-deoxy-
-D-galactopyranoside (GalNAc-O-bn) leads to marked effects upon the phenotypic properties of mucin-secreting or enterocyte-like HT-29 cells: an inhibition in the secretion of mucins, a blockade in the apical targeting of membrane brush border glycoproteins and a swelling of cells with intracellular accumulation of numerous vesicles. Folch extraction and partition of treated enterocyte-like HT-29 cells revealed a very important accumulation of orcinol and/or resorcinol reactive material in the upper phase (usually containing gangliosides), as compared with untreated HT-29 cells and with treated and untreated Caco-2 cells. Structural analysis indicated the accumulation of a series of GalNAc-O-bn derived oligosaccharides, most of them with the common core Galß1-3GalNAc-O-bn. These oligosaccharides contained residues of GlcNAc, Gal, Neu5Ac, or Fuc. In particular, the tagged sialyl-Lewisx was identified, as well as more complex sialylated derivatives, including the sialyl-Lewisx substituted by an additional Neu5Ac residue. The benzylated oligosaccharides were not significantly detected in the culture medium except for Galß1–3GalNAc-O-bn. Upon reversion of the treatment, these derivatives disappeared from the cells within few days, however were not recovered as such in the culture medium. Intracellular degradation occurred with desialylation and defucosylation as the first steps. The spectacular accumulation of benzylated oligosaccharides in HT-29 cell, permanently exposed to GalNAc-O-bn very likely plays an important role in the alterations of cellular processes previously described in this cell line. The HT-29 cell culture system also appears to be an efficient source of several tagged oligosaccharides.
1 To whom correspondence should be addressed at: Unité de Biologie et Physiopathologie des Cellules Mucipares, INSERM U-377, place de Verdun, F-59045 Lille, France
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  G. Patsos, V. Hebbe-Viton, C. Robbe-Masselot, D. Masselot, R. S. Martin, R. Greenwood, C. Paraskeva, A. Klein, M. Graessmann, J. C. Michalski, et al. O-Glycan inhibitors generate aryl-glycans, induce apoptosis and lead to growth inhibition in colorectal cancer cell lines Glycobiology, April 1, 2009; 19(4): 382 - 398. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Julien, E. Adriaenssens, K. Ottenberg, A. Furlan, G. Courtand, A.-S. Vercoutter-Edouart, F.-G. Hanisch, P. Delannoy, and X. Le Bourhis ST6GalNAc I expression in MDA-MB-231 breast cancer cells greatly modifies their O-glycosylation pattern and enhances their tumourigenicity Glycobiology, January 1, 2006; 16(1): 54 - 64. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Delacour, V. Gouyer, J.-P. Zanetta, H. Drobecq, E. Leteurtre, G. Grard, O. Moreau-Hannedouche, E. Maes, A. Pons, S. Andre, et al. Galectin-4 and sulfatides in apical membrane trafficking in enterocyte-like cells J. Cell Biol., May 9, 2005; 169(3): 491 - 501. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Delacour, V. Gouyer, E. Leteurtre, T. Ait-Slimane, H. Drobecq, C. Lenoir, O. Moreau-Hannedouche, G. Trugnan, and G. Huet 1-Benzyl-2-acetamido-2-deoxy-{alpha}-D-galactopyranoside Blocks the Apical Biosynthetic Pathway in Polarized HT-29 Cells J. Biol. Chem., September 26, 2003; 278(39): 37799 - 37809. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Ulloa and F. X. Real Benzyl-N-acetyl-alpha -D-galactosaminide Induces a Storage Disease-like Phenotype by Perturbing the Endocytic Pathway J. Biol. Chem., March 28, 2003; 278(14): 12374 - 12383. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Leteurtre, V. Gouyer, D. Delacour, B. Hemon, A. Pons, C. Richet, J.-P. Zanetta, and G. Huet Induction of a Storage Phenotype and Abnormal Intracellular Localization of Apical Glycoproteins Are Two Independent Responses to GalNAc{alpha}-O-bn J. Histochem. Cytochem., March 1, 2003; 51(3): 349 - 361. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-P. Zanetta, A. Pons, M. Iwersen, C. Mariller, Y. Leroy, P. Timmerman, and R. Schauer Diversity of sialic acids revealed using gas chromatography/mass spectrometry of heptafluorobutyrate derivatives Glycobiology, August 1, 2001; 11(8): 663 - 676. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Ulloa and F. X. Real Differential Distribution of Sialic Acid in {{alpha}}2,3 and {{alpha}}2,6 Linkages in the Apical Membrane of Cultured Epithelial Cells and Tissues J. Histochem. Cytochem., April 1, 2001; 49(4): 501 - 510. [Abstract] [Full Text]
|
|
|
|
 |
|
 V Gouyer, E Leteurtre, P Delmotte, W. Steelant, M. Krzewinski-Recchi, J. Zanetta, T Lesuffleur, G Trugnan, P Delannoy, and G Huet Differential effect of GalNAc(&agr;)-O-bn on intracellular trafficking in enterocytic HT-29 and Caco-2 cells: correlation with the glycosyltransferase expression pattern J. Cell Sci., January 4, 2001; 114(8): 1455 - 1471. [Abstract] [PDF]
|
|
|
|
|
|
H. Hassan, C. A. Reis, E. P. Bennett, E. Mirgorodskaya, P. Roepstorff, M. A. Hollingsworth, J. Burchell, J. Taylor-Papadimitriou, and H. Clausen The Lectin Domain of UDP-N-acetyl-D-galactosamine:Polypeptide N-acetylgalactosaminyltransferase-T4 Directs Its Glycopeptide Specificities J. Biol. Chem., December 1, 2000; 275(49): 38197 - 38205. [Abstract] [Full Text] [PDF]
|
|