Abstract
Anthrax infections progress at a rapid pace, making rapid detection methods of utmost importance. MALDI-MS proteomics methods focused on Bacillus anthracis detection have targeted chromosomally encoded proteins, which are highly conserved between closely related species, hindering species identification. Presented here is an AP-MALDI-MS method targeting plasmid-borne proteins from Bacillus spores for species-level identification. A bioinformatics analysis revealed that 60.3% and 75.4% of tryptic peptides from plasmid-borne proteins of B. anthracis and B. thuringiensis were species-specific, respectively. Reported here is a method in which plasmid-borne Δ-endotoxins were extracted directly from B. thuringiensis spores in 100 mM KOH. The pH was then adjusted to 8 and a 5-min trypsin digestion was performed on the extracted proteins. The resulting tryptic peptides were analyzed by AP-MALDI-MS/MS, which produced a definitive identification the B. thuringiensis speciesspecific Cry1Ab protein with a MASCOT score of 278 and expect value of 7.5 × 10−23. This method has demonstrated the detection and identification of B. thuringiensis spores at the species level following a 5-min trypsin digestion. The challenges in applying a similar approach to the detection of plasmid-borne protein toxins from B. anthracis are also discussed.
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Nguyen, J., Russell, S.C. Targeted proteomics approach to species-level identification of Bacillus thuringiensis spores by AP-MALDI-MS. J Am Soc Mass Spectrom 21, 993–1001 (2010). https://doi.org/10.1016/j.jasms.2010.01.032
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DOI: https://doi.org/10.1016/j.jasms.2010.01.032