Fine molecular analysis of the antigenicity of the Androctonus australis hector scorpion neurotoxin II: a new antigenic epitope disclosed by the Pepscan method.

A set of 58 overlapping rod-bound peptides was used to map the antigenic reactivity pattern of a 64-residue neurotoxin (AaH II) from the venom of the scorpion Androctonus australis hector. Five anti-toxin rabbit antisera were assayed serially for their capacity to bind to each peptide in the set. Six regions of antigenic reactivity were thus identified (sequences: 1-8, 4-12, 27-35, 39-45, 52-58 and 55-61). When positioned on a 3-D model of the toxin, these regions appeared to correspond to either beta-turn or extended parts of the molecule. The antigenic regions revealed by this technique agree fairly well with those previously mapped on the same toxin by different methods. One discrepancy was, however, that the present study shows the N-terminus to be strongly reactive with anti-toxin antibodies. The antigenicity of this region was confirmed, since rabbit antibodies raised against a synthetic peptide mimicking the sequence 1-8 of the toxin were found to bind the toxin with high efficiency. A fine analysis of the recognition of this region was performed. Alanine-containing analogs of the sequence 1-7 and peptides mimicking the N-terminal of the four main toxins of AaH were probed with anti-toxin and anti-peptide antibodies. Lysine 2, aspartic acid 3 and glycine 4 were shown to be key residues in the recognition of the N-terminal region of the AaH II toxin by anti-toxin antibodies. In contrast, a loose specificity of recognition was shown by one anti-peptide serum which was, in addition, able to recognize the N-termini of all four AaH toxins.