Carbohydrate changes in glycoproteins of a poorly metastasizing wheat germ agglutinin-resistant melanoma clone.
Glycoproteins of a metastasizing line of B16 mouse melanoma and a poorly metastasizing wheat germ agglutinin-resistant clone were compared. Cell surface proteins and glycoproteins were isotopically labeled by lactoperoxidase-catalyzed iodination and by NaB3H4 reduction after oxidation by periodate or galactose oxidase and subsequently analyzed by gel electrophoresis and autoradiography. Differences were observed in the relative mobilities of several major cell surface components. Binding of 125I-labeled lectins to total cellular proteins on polyacrylamide gels following electrophoresis showed that the major wheat germ agglutinin-binding components of F1 cells were altered in Wa-4 cells. Similar differences were not observed in concanavalin A-binding components. Total cellular glycopeptides were analyzed after separation into structurally distinct classes. The acidic "complex" N-glycosidic glycopeptides from the resistant cells were of lower molecular weight than those from the parent cells. No differences were observed among the mannose-rich N-glycosidic glycopeptides or the alkali-labile O-glycosidic oligosaccharides. Structural studies involving methylation analysis revealed that in the altered glycopeptides of the resistant cells the amount of neuraminic acid residues was decreased to one-half, concomitant with an increase in the amount of fucose. The lost sialic acid was bound to C-3 of galactose, whereas the increased fucose was found on C-3 of 4-substituted N-acetylglucosamine. A possible basis for the glycosylation change and its relation to the biological behavior are discussed.
D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002462:Cell Membrane / Q000378:metabolism; D002850:Chromatography, Gel; D002999:Clone Cells; D004591:Electrophoresis, Polyacrylamide Gel; D006020:Glycopeptides / Q000032:analysis / Q000302:isolation & purification; D006023:Glycoproteins / Q000378:metabolism*; D037102:Lectins; D008545:Melanoma / Q000378:metabolism*; D051379:Mice; D009362:Neoplasm Metastasis; D009374:Neoplasms, Experimental / Q000378:metabolism; D011485:Protein Binding
D002241:Carbohydrates; D006020:Glycopeptides; D006023:Glycoproteins; D037102:Lectins