Partial characterization of sialoglycopeptides produced by cultured human melanoma cells and melanocytes.
The sialoglycopeptides produced by HM7 human melanoma and fetal uveal melanocyte cultures grown in the presence of [3H]glucosamine and [35S]sulfate were isolated from the Pronase digests of cells, spent media, and intracellular material. From the melanoma culture, six sialoglycopeptides, accounting for 43% of the total 3H radioactivity in the nondiffusible cell-associated glycopeptides, were purified. A major glycopeptide (GPIb) having an apparent molecular weight in the range 12 000-15 000 showed specific sialic acid dependent to interaction with wheat germ agglutinin (WGA). It was found to contain mainly O-glycosidically linked oligosaccharides having the structure (AcNeu) leads to 0-2[Gal leads to GalNAc]; some N-glycosidically linked saccharides were also present. A second WGA-binding glycopeptide (GPIa) was smaller and less anionic and had a higher proportion of N-glycosidically linked saccharides than GPIb. The normal fetal cultures yielded either no (iris) or markedly reduced (melanocytes) quantities of the WGA-binding glycopeptides. The four WGA-nonbinding sialoglycopeotides purified from melanoma were shown to have complex (N-acetyllactosaminly type) oligosaccharides linked via N-acetylglucosamine to asparagine with either no or insignificant amounts of O-glycosidically linked saccharides. The corresponding glycopeptides from melanocytes were of smaller molecular size and lower anionic charge, reflecting an overall lower degree of glycosylation.
D002460:Cell Line; D005260:Female; D005333:Fetus; D006595:Hexosamines / Q000032:analysis; D006801:Humans; D008544:Melanocytes / Q000378:metabolism*; D008545:Melanoma / Q000378:metabolism*; D008970:Molecular Weight; D009439:Neuraminidase; D011247:Pregnancy; D012794:Sialic Acids / Q000032:analysis; D012795:Sialoglycoproteins / Q000096:biosynthesis / Q000302:isolation & purification*; D014734:Vibrio cholerae / Q000201:enzymology
D006595:Hexosamines; D012794:Sialic Acids; D012795:Sialoglycoproteins; D009439:Neuraminidase