A deleted variant of Bacillus anthracis protective antigen is non-toxic and blocks anthrax toxin action in vivo.
Anthrax toxin is the only protein secreted by Bacillus anthracis that contributes to the virulence of this bacterium. An obligatory step in the action of anthrax toxin on eukaryotic cells is cleavage of the receptor-bound protective antigen (PA) protein (83 kilodaltons) to produce a 63-kilodalton, receptor-bound COOH-terminal fragment. A similar fragment can be obtained by limited treatment with trypsin. This proteolytic processing event exposes a site with high affinity for the other two anthrax toxin proteins, lethal factor and edema factor. Terminal sequencing of the purified fragment showed that the activating cleavage occurred in the sequence Arg164-Lys165-Lys166-Arg167. The gene encoding PA was mutagenized to delete residues 163-168, and the deleted PA was purified from a Bacillus subtilis host. The deleted PA was not cleaved by either trypsin or the cell-surface protease, and was non-toxic when administered with lethal factor. Purified, deleted PA protected rats when administered 90 min before injection of 20 minimum lethal doses of toxin. This mutant PA may be useful as a replacement for the PA that is the major active ingredient in the current human anthrax vaccine, because deleted PA is expected to have normal immunogenicity, but would not combine with trace amounts of LF and EF to cause toxicity.
Singh
Y
Y
Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21701-5011.
Chaudhary
V K
VK
Leppla
S H
SH
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D001408:Bacillus anthracis / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D014644:Genetic Variation; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids; D014774:Virulence
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; C030325:anthrax toxin