Serum-mediated cleavage of Bacillus anthracis protective antigen is a two-step process that involves a serum carboxypeptidase

Much of our understanding of the activity of anthrax toxin is based on in vitro systems, which delineate the interaction between Bacillus anthracis toxins and the cell surface. However, these systems fail to account for the intimate association of B. anthracis with the circulatory system, including the contribution of serum proteins to the host response and processing of anthrax toxins. Using a variety of immunological techniques to inhibit serum processing of B. anthracis protective antigen (PA) along with mass spectrometry analysis, we demonstrate that serum digests PA via 2 distinct reactions. In the first reaction, serum cleaves PA₈₃ into 2 fragments to produce PA₆₃ and PA₂₀ fragments, similarly to that observed following furin digestion. This is followed by carboxypeptidase-mediated removal of the carboxyterminal arginine and lysines from PA₂₀.