Presence of terminal N-acetylgalactosamine residues in subregions of the endoplasmic reticulum is influenced by cell differentiation in culture.
Using Helix pomatia lectin as a specific probe for terminal, nonreducing N-acetylgalactosamine residues, glycoprotein precursors bearing newly initiated O-linked oligosaccharides have been localized in the lumen of the endoplasmic reticulum and cis-Golgi cisternae. This pattern contrasts with the detection of the terminal disaccharide galactose beta-1,3-N-acetylgalactosamine by Arachis hypogaea lectin in middle and trans-Golgi compartments, which are considered elongation sites for O-glycosylation. Distribution of H. pomatia ligands in the endoplasmic reticulum is confined to specialized regions or subcompartments in both human colonic adenocarcinoma cells and cultured chicken chondrocytes. Since in cartilage, chondrocytes contain H. pomatia-binding sites exclusively concentrated in cis-Golgi cisternae, primary cultures of this cell type have been used to study those conditions that promote initiation of O-glycosylation in the endoplasmic reticulum. A correlation has been found between the age of the culture and the extent of reactivity of the endoplasmic reticulum with either H. pomatia lectin or antibody against the sequence GalNAc alpha-serine/threonine (Tn antigen). Cells showing an extensive reaction are not hindered in their secretory activity and still maintain the chondrocyte phenotype. Taken together the results suggest that the intracellular distribution of the glycosylation enzymes is not only cell type-specific as previously shown (Roth, J., Taatjes, D. J., Weinstein, J., Paulson, J. C., Greenwell, P., and Watkins, W. M. (1986) J. Biol. Chem. 261, 14307-14312) but it might also vary depending on the stage of cell differentiation.
Perez-Vilar
J
J
Department of Cell Biology, Faculty of Biology, University of Seville, Spain.
Hidalgo
J
J
Velasco
A
A
D000116:Acetylgalactosamine / Q000032:analysis*; D000230:Adenocarcinoma; D000818:Animals; D002356:Cartilage / Q000166:cytology* / Q000648:ultrastructure; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D002642:Chick Embryo; D003110:Colonic Neoplasms; D004721:Endoplasmic Reticulum / Q000648:ultrastructure*; D006031:Glycosylation; D006056:Golgi Apparatus / Q000648:ultrastructure; D006372:Helix, Snails; D006801:Humans; D037102:Lectins
C038095:Helix lectin; D037102:Lectins; D000116:Acetylgalactosamine