Structure of tetanus toxin. II. Toxin binding to ganglioside.
The interaction between tetanus toxin and ganglioside containing 2 N-acetylneuraminic acid residues linked in sequence to one another has been investigated using a new method involving radioactively labeled ganglioside and tetanus toxin adsorbed to Sephadex matrix. Binding between the two components was demonstrated, and it was calculated that in the nanomolar concentration range, tetanus toxin becomes half-saturated at about 5 X 10(-8) M concentration of ganglioside. Removal of the ceramide portion from the ganglioside resulted in the complete loss of binding activity, whereas removal of the terminal N-acetylneuraminic acid residue from the intact ganglioside had no effect. Among the fragments derived from tetanus toxin (Helting, T. B., and Zwisler, O. (1977) J. Biol. Chem. 252, 187-193), only the heavy chain polypeptide exhibited a binding activity of the same order of magnitude as that observed for the native toxin. The light chain polypeptide showed no interaction with ganglioside and among the fragments derived from the toxin by digestion with papain, only Fragment C, at a high protein concentration, displayed marginal binding activity. Using monovalent antibodies directed against specific regions of the tetanus toxin molecule, it was demonstrated that antibodies directed against Fragment C uniquely interfere with the binding process. Anti-light chain serum was ineffective, as well as antitetanus toxoid serum previously absorbed with Fragment C. It is concluded that the binding site for ganglioside is located on the heavy chain portion of tetanus toxin, possibly in or near the region comprised by Fragment C.
D001665:Binding Sites; D000939:Epitopes; D005732:Gangliosides / Q000378:metabolism; D007700:Kinetics; D010446:Peptide Fragments; D011485:Protein Binding; D012685:Sepharose; D012794:Sialic Acids; D013743:Tetanus Antitoxin; D013744:Tetanus Toxin
D000939:Epitopes; D005732:Gangliosides; D010446:Peptide Fragments; D012794:Sialic Acids; D013743:Tetanus Antitoxin; D013744:Tetanus Toxin; D012685:Sepharose