Monoclonal antibodies against Dictyostelium plasma membranes: their binding to simple sugars

Abstract

Monoclonal antibodies raised against purified membranes from Dictyostelium discoideum were classified according to three criteria: type of antigen as revealed in immunoblots, developmental regulation of the target antigens, and location of the antigens on the cell surface. Some antibodies reacted with myosin, two with glycolipids. One group of antibodies bound to the protein moiety of the contact site A glycoprotein, whereas another group reacted with carbohydrate epitopes that the contact site A glycoprotein shared with a few other membrane glycoproteins. Binding of the latter antibodies to their antigens was either specifically blocked by N-acetylglucosamine or by maltose as well as methyl-alpha-mannoside and N-acetylglucosamine. These anti-carbohydrate antibodies bound specifically to agarose beads derivatized with some sugars. These results and competition studies with several carbohydrates suggest that the epitope recognized by the antibodies contains as major components N-acetylglucosamine, maltose and alpha-mannose residues. One monoclonal antibody, which reacts with N-acetylglucosamine, was used for affinity purification of the contact site A glycoprotein from a crude membrane extract. N-acetylglucosamine was used as a mild eluent of the antigen from the antibody column. No detergents were added during the entire purification procedure.