Differentiation-dependent glycosylation of gp190, an oncofetal crypt cell antigen expressed by Caco-2 cells.
gp190 is a glycoprotein expressed on the cell surface of several human colon carcinoma cells in culture, on epithelial cells of fetal colon, but not on the normal mucosa of adult colon; thus it is referred to as an oncofetal crypt cell antigen. We report the characterisation of O-linked glycans carried by gp190 synthesised by [3H]glucosamine-labelled Caco-2 cells at the confluence (undifferentiated cells) and at three weeks of postconfluence (differentiated cells). By using a specific monoclonal antibody, gp190 was isolated and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The mobility of gp190 from differentiated cells was found to be lower than that from undifferentiated cells, suggesting a more extensive glycosylation process in the former glycoprotein. The major results of the glycan characterisation have been as follows: (i) gp190 carries mainly, if not exclusively, O-linked glycans with the core-2 structure; (ii) the elongation with N-acetyllactosamine units of the Gal beta1,4GlcNAc beta1,6(Gal beta1,3)GalNAc tetrasaccharide predominates in gp190 synthesised by differentiated cells, whereas the direct alpha2,3sialylation of the tetrasaccharide is prevalent in gp190 synthesised by undifferentiated cells. The increment in the core-2 beta1,6GlcNAc-transferase activity under the Caco-2 differentiation process may be relevant in producing the larger occurrence of polylactosaminoglycans in gp190 from differentiated cells. Since no change in the activity of the alpha2,3sialyltransferases upon cell differentiation was observed, we suggest that the lower alpha2,3sialylation in gp190 synthesised by polarised cells might be due to a changed transit-rate through the distal Golgi apparatus.
Department of Experimental Pathology, University of Bologna, Italy.
N
N
Malagolini
D
D
Cavallone
F
F
Serafini-Cessi
D000606:Amino Sugars / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000302:isolation & purification / Q000378:metabolism*; D018938:Caco-2 Cells; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002454:Cell Differentiation; D005591:Chemical Fractionation; D006031:Glycosylation; D016695:Glycosyltransferases / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry; D012794:Sialic Acids / Q000737:chemistry
D000606:Amino Sugars; D000951:Antigens, Neoplasm; D011134:Polysaccharides; D012794:Sialic Acids; C031192:oncofetal antigens; C000458:N-acetyllactosamine; D016695:Glycosyltransferases