Investigation of the Carbohydrate Structures of Total Cellular and Secreted Glycoproteins from Jar Cells
Loesel, Laura L.
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A human malignant cell line from choriocarcinoma (JAR) labelled with D-[6- 3H] was available in the Ruddon pharmacology laboratory, at the University of Michigan. The glycoproteins, human chorionic gonadotropin (hCG) and laminin, had been previously removed from the cell lysates and culture media by "immunoprecipitation. The oligosaccharide structures of these two glycoproteins were characterized. Initial investigations showed different glycosylation patterns and structures for these two glycoproteins, suggesting selective cellular processing. However, hCG and laminin represent approximately 1% of the total glycoprotein content of JAR cells. This project is an investigation of the structure of the N-linked glycans from the remaining total glycoprotein population in the JAR cell line. Characterization of the oligosaccharides found in the total glycoprotein population will be accomplished using several techniques: ethanol precipitation of the total glycoproteins, enzymatic cleavage of N-linked oligosaccharides, high performance liquid chromatography, gel filtration chormatography, and lectin affinity chromatography.