This proposal represents a continuation of our studies on the structure and metabolism of cell surfaces glycoconjugates with special emphasis on Ehrlich ascites tumor (EAT) cells and activated murine peritoneal macrophages. It is proposed to continue study of the biosynthesis and degradation of the family of unique alpha-D-galactosyl-terminated glycoproteins which occur on the surface of EAT cells. Isolation and characterization of the biosynthetic and degradative enzymes will be undertaken. Similarly, structural and biosynthetic studies of a specific macrophage activation glycoprotein (MAG) will be conducted. In addition, structural studies on the carbohydrate residues of a basement membrane glycoprotein of great biological importance - laminin, of murine and human origin, are in progress. The methodologies to be employed in these investigations include chemical and physical techniques of carbohydrate chemistry, affinity chromatographic isolation of glycosyltransferases, lectin binding and precipitation studies, generation of clonal variants of EAT cells and the production of monoclonal antibodies against glycoproteins and defined sequences of carbohydrate, e.g. the Ga1 alpha1, 3Ga1 beta1, 4G1cNAc trisaccharide which occurs on the surface of EAT cells and on the laminin molecule. Using this monoclonal antibody we shall attempt to confer passive immunity in mice against Ehrlich ascites tumor cells.
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