Comparative study of glycosyltransferase activities in Caco-2 cells before and after enterocytic differentiation using lectin-affinity high-performance liquid chromatography.
Human colonic adenocarcinoma Caco-2 cells differentiate into enterocytes by induction with sodium butyrate after confluence. Our previous studies have shown that there are high levels of H type 1 blood group antigen and core 2 structure present in O-glycans of the glycoproteins from these differentiated cells and these O-glycans appear to be indispensable for the process of differentiation of the cells (J. Amano and M. Oshima, 1999, J. Biol. Chem. 274, 21209-21216). Here, we have determined the glycosyltransferase activities using lectin-affinity HPLC because the method enabled easy separation and identification of mixtures of isomeric oligosaccharide structures due to the high resolution and reproducibility. The activities of beta 3-galactosyltransferase, alpha 2-fucosyltransferase, which are responsible for H type 1 antigen biosynthesis, and core 2 beta 6-N-acetylglucosaminyltransferase in differentiated Caco-2 cells were higher than those in undifferentiated cells. These results demonstrate that an increase in specific glycosyltransferase activities brought on a change of the O-glycan structures during differentiation.
Amano
J
J
Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033, Japan. amano@mol.f.u-tokyo.ac.jp
Kobayashi
K
K
Oshima
M
M
D000818:Animals; D018938:Caco-2 Cells; D002454:Cell Differentiation; D002851:Chromatography, High Pressure Liquid; D020895:Enterocytes / Q000166:cytology*; D005690:Galactose / Q000378:metabolism; D005700:Galactosyltransferases / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D016695:Glycosyltransferases / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D037102:Lectins / Q000378:metabolism*; D008956:Models, Chemical; D009237:N-Acetyllactosamine Synthase / Q000378:metabolism; D011485:Protein Binding; D013379:Substrate Specificity; D013997:Time Factors; D014407:Tumor Cells, Cultured
D037102:Lectins; D016695:Glycosyltransferases; D005700:Galactosyltransferases; C068666:globoside galactosyltransferase; D009237:N-Acetyllactosamine Synthase; D006026:Glycoside Hydrolases; D005690:Galactose