Analysis of glycoprotein-bound carbohydrates from pluripotent embryonal carcinoma cells by pokeweed agglutinin-agarose.
Embryoglycan is the high-molecular-weight poly-N-acetyllactosamine characteristically and abundantly present in early embryonic cells. Among lectins reacting with poly-N-acetyllactosamines pokeweed agglutinin (PWA) was found to be most useful in analyzing glycoproteins from HM-1 pluripotent embryonal carcinoma (EC) cells, since virtually all glycoproteins carrying embryoglycan in these cells could be isolated by affinity chromatography on PWA-agarose. Furthermore almost all of embryoglycan from HM-1 cells bound to PWA-agarose. Since PWA-agarose used for the present study was confirmed to bind branched but not linear poly-N-acetyllactosamines, the above result indicated that embryoglycan lacking branched poly-N-acetyllactosaminyl chain was scarcely present in these cells. The same approach was used as a mean to show that embryoglycan with Lotus tetragonolobus agglutinin receptor activity also usually has the branched poly-N-acetyllactosamine structure in EC cells. Glycoprotein fractions from PYS-2 parietal endoderm cells and from STO fibroblasts also bound to PWA-agarose. However, the ratio of PWA binding fraction to the total [14C]galactose-labeled glycoproteins was less in these cells as compared to the value in EC cells, and the poly-N-acetyllactosamines from these cells exhibited lower molecular weights than embryoglycan. These results are consistent with our proposal that the complexity and abundance of poly-N-acetyllactosamines distinguishes EC cells from most other cells.
Muramatsu
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H
Department of Biochemistry, Faculty of Medicine, Kagoshima University.
Muramatsu
T
T
D000818:Animals; D002847:Chromatography, Agarose / Q000379:methods; D054278:Embryonal Carcinoma Stem Cells; D005690:Galactose / Q000378:metabolism; D006023:Glycoproteins / Q000032:analysis*; D008745:Methylation; D051379:Mice; D014411:Neoplastic Stem Cells / Q000032:analysis*; D011043:Pokeweed Mitogens / Q000032:analysis*; D011134:Polysaccharides / Q000378:metabolism; D011975:Receptors, Mitogen / Q000302:isolation & purification
D006023:Glycoproteins; D011043:Pokeweed Mitogens; D011134:Polysaccharides; D011975:Receptors, Mitogen; C045474:embryoglycan; C037199:poly-N-acetyllactosamine; D005690:Galactose