Inhibition of the glycosylation and alteration in the intracellular trafficking of mucins and other glycoproteins by GalNAca-O-bn in mucosal cell lines: An effect mediated through the intracellular synthesis of complex GalNAca-O-bn oligosaccharides

To address the function of carbohydrates in mucins, GalNAc alpha -O-bn has been used in in vivo experiments on several human mucosal cultured cells as a potential competitor of the glycosylation of N-acetylgalactosamine resid ues. GalNAc alpha -O-bn is metabolized by glycosyltransferases expressed in the cell, and give rise to different internal derivatives starting in part icular from the formation of the disaccharide Gal beta1-3GalNAc alpha -O-bn . In this line, GalNAc alpha -O-bn exposure inhibits peripheral glycosylati on according a cell-type specific manner. The metabolic alterations are ver y important in HT-29 cell line, leading to a massive accumulation of GalNAc alpha -O-bn oligosaccharide derivatives and to a strong inhibition of the terminal elongation of O-glycans by alpha2,3 sialyltransferase ST3Gal I. Ga lNAc alpha -O-bn treatment also induced alterations at the cellular level, exhibiting a large scale in HT-29 cells, i.e. 1) an inhibition of mucin sec retion, 2) a blockade in the targeting of some membrane glycoproteins (brus h border glycoproteins such as dipeptidylpeptidase IV, carcinoembryonic ant igen and the mucin-like glycoprotein MUC1, and the basolateral cell adhesio n molecule CD44), 3) an inhibition in the processing of lysosomal enzymes. Morphological abnormalities have been evidenced in GalNAc alpha -O-bn treat ed cells, in particular the accumulation of numerous intracellular vesicles in HT-29 cells. Taken together, these data suggest that O-glycosylation mi ght be involved in the regulation of the targeting of O-glycosylproteins th rough carrier vesicles.