“…Peptides were sequenced on a Shimadzu PPSQ-10 gas-phase sequencer either without prior derivatization or after reduction and 4-vinyl-pyridine alkylation [4]. Reduced alkylated toxins were submitted to proteolysis by (a) TPCK-treated trypsin (Sigma Co., USA), 2% w/w, 37³C, 14 h in 100 mM ammonium bicarbonate, 0.1 mM CaCl 2 pH 8.1 (for Bm32-VI, a second digestion was performed with 2% w/w of enzyme added after 4 h and a total reaction time of 24 h), (b) asparagine endopeptidase (Asp-N, Takara Shuzo, Japan), 5% w/w, 37³C, 14 h in 50 mM sodium phosphate bu¡er pH 8.0, and (c) Achromobacter protease I (API, Wako, Japan), 5% w/w, 37³C, 14 h in 100 mM Tris bu¡er, pH 8.0 and 12.5%, 37³C, 14 h in 100 mM ammonium bicarbonate pH 8.0.…”