L2/HNK-1 carbohydrate and protein-protein interactions mediate the homophilic binding of the neural adhesion molecule P0.
The neural adhesion molecule P0, the most abundant glycoprotein in peripheral myelin of mammals, is a member of the immunoglobulin superfamily and expresses the L2/HNK-1 and L3 oligosaccharides at a single N-glycosylation site. It acts in both homophilic and heterophilic binding mechanisms. To investigate the molecular requirements for homophilic interaction, we have used P0 from human sciatic nerve and the extracellular domain of P0 expressed in bacteria to determine binding of P0 to P0 in solid phase and bead aggregation assays. The binding of P0 to P0 could be partially inhibited in both assays by antibodies to the L2/HNK-1 epitope and by the L2/HNK-1 carbohydrate, but not by L3 antibodies or other carbohydrates. Inhibition of binding was also seen with polyclonal antibodies reacting with the protein backbone of P0. These observations indicate that both carbohydrate and protein structures are involved in the binding of P0 to P0 and that P0 acts as a presenter of and a receptor for a functionally important carbohydrate.
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation / Q000378:metabolism*; D018959:CD57 Antigens; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism*; D005732:Gangliosides / Q000378:metabolism; D006017:Glycolipids / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D018993:Myelin P0 Protein; D009185:Myelin Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D011485:Protein Binding; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012584:Sciatic Nerve / Q000378:metabolism; D013433:Sulfoglycosphingolipids / Q000378:metabolism
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D018959:CD57 Antigens; D015816:Cell Adhesion Molecules, Neuronal; D005732:Gangliosides; D006017:Glycolipids; D046911:Macromolecular Substances; D018993:Myelin P0 Protein; D009185:Myelin Proteins; D009844:Oligosaccharides; D011994:Recombinant Proteins; D013433:Sulfoglycosphingolipids; C033851:ganglioside, GD1b