Polyglycosylceramides with branched N-acetyllactosamine sequences are synthesized by the human pancreatic carcinoma cell line PANC-1.
We have metabolically labeled the human pancreatic tumor cell line PANC-1 with high specific activity tritiated sugar precursors to study the expression of glycosphingolipids by this cell type. We have used a combination of detergent solubilization, exhaustive protease digestion, ceramide glycanase digestion, and reverse-phase chromatography to isolate glycosphingolipid-derived oligosaccharides specifically labeled in their component sugars. A significant proportion of the oligosaccharides derived from polar glycosphingolipids were of high molecular mass (greater than 2000 Da). The results of compositional studies, lectin affinity chromatography, and methylation analysis suggested that this high molecular weight fraction consists of lactosaminoglycan type oligosaccharides derived from polyglycosylceramides. There are on average three beta 1-6 linked N-acetyllactosamine branches attached to the polylactosamine backbone in this type of glycosphingolipid-derived oligosaccharide. The majority of the oligosaccharides also contain 1-2 mol of sialic acid that are linked alpha 2-3 to penultimate galactose. The results indicate that PANC-1 cells, like human colorectal tumor cells, express highly extended neolacto type glycosphingolipids. However, the lactosaminoglycan sequences are highly branched, unlike those associated with colorectal tumor cells.
Barnett
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Department of Biochemistry, Eastern Virginia Medical School, Norfolk 23501.
Clark
G F
GF
D000606:Amino Sugars / Q000378:metabolism*; D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid; D002855:Chromatography, Thin Layer; D006028:Glycosphingolipids / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008745:Methylation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000606:Amino Sugars; D006028:Glycosphingolipids; D037102:Lectins; D009844:Oligosaccharides; C000458:N-acetyllactosamine