Anti-idiotypic monoclonal antibodies to GM1 identify ganglioside binding proteins.
Ganglioside stimulated neurite outgrowth may be due to ganglioside binding to membrane proteins or to intercalation into the membrane. To test that ganglioside binding proteins could be found on neuronal surfaces, anti-idiotypic ganglioside monoclonal antibodies (AIG mAbs) were generated to mimic the biological properties of the GM1 ganglioside. The AIG mAbs were identified by their ability to bind to a known GM1 binding protein, the beta-subunit of cholera toxin. For the two AIG mAbs studied, AIG5 and AIG20, binding to beta-CT was blocked most strongly by GM1. This data also suggests that AIG5 and AIG20 mimic different but overlapping epitopes of the ganglioside GM1. Western blotting and immunoprecipitation of mammalian tissues reveals four potential ganglioside binding proteins of molecular weight 93, 66, 57, and 45 kDa. Immunocytochemistry demonstrates neuronal surface label with the AIG mAbs, which suggests that gangliosides, enriched on the neuronal surface membrane, are co-localized with putative ganglioside binding proteins. In bioassays, the AIG mAbs promote neuronal sprouting. This shows that these antibodies can be used to study the biological effects of ganglioside binding to neuronal surface proteins, and the role of gangliosides in the activation of neurite outgrowth.
Riggott
M J
MJ
Department of Neurobiology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Matthew
W D
WD
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D005260:Female; D005677:G(M1) Ganglioside / Q000276:immunology* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding; D051381:Rats
D000888:Antibodies, Anti-Idiotypic; D000941:Antigens; D008565:Membrane Proteins; D005677:G(M1) Ganglioside