Characterization of oligosaccharides that bind to human anti-MAG antibodies and to the mouse monoclonal antibody HNK-1.
In some patients with neuropathy and IgM M-proteins the M-proteins bind to a carbohydrate determinant that is shared by the CNS and PNS myelin-associated glycoprotein (MAG) and by several additional glycoproteins and 2 glycolipids in peripheral nerve. The HNK-1 mouse monoclonal antibody binds to the same glycoproteins and glycolipids as well as to a number of other neuronal adhesion molecules and to human natural killer cells. To isolate the epitope-bearing oligosaccharides from their respective glycoproteins we digested delipidated spinal cord and peripheral nerve with pronase. The resulting glycopeptides were fractionated by concanavalin A-Sepharose chromatography to yield tri- and tetraantennary-complex, biantennary-complex and high mannose-type glycopeptides. Glycopeptides bearing the antigenic determinant were identified by their ability to block binding of M-proteins and HNK-1 antibodies to MAG-coated microwells by enzyme-linked immunosorbent assay (ELISA). Blocking activity was detected in the tri- and tetraantennary glycopeptide fraction from both CNS and PNS. The blocking activity was destroyed by pretreatment of the isolated glycopeptides with mild acid hydrolysis. Further fractionation by gel filtration chromatography indicated that the reactive glycopeptides from peripheral nerve and spinal cord eluted in the same position. The data suggest that CNS and PNS MAG and other peripheral nerve glycoproteins share similar oligosaccharides, and that the M-proteins and HNK-1 bind to the same structures.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D001798:Blood Proteins / Q000276:immunology / Q000378:metabolism*; D001923:Brain Chemistry; D003711:Demyelinating Diseases / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D007136:Immunoglobulins; D051379:Mice; D009185:Myelin Proteins / Q000276:immunology*; D018992:Myelin-Associated Glycoprotein; D009844:Oligosaccharides / Q000302:isolation & purification / Q000378:metabolism*; D010525:Peripheral Nerves / Q000032:analysis; D013116:Spinal Cord / Q000032:analysis
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D006023:Glycoproteins; D007136:Immunoglobulins; C009823:M-proteins (Myeloma); D009185:Myelin Proteins; D018992:Myelin-Associated Glycoprotein; D009844:Oligosaccharides