Publication Date:
2012-03-11
Description:
Botulinum neurotoxins (BoNTs) cause the disease botulism, which can be lethal if untreated. There are seven known serotypes of BoNT, A–G, defined by their response to antisera. Many serotypes are distinguished into differing subtypes based on amino acid sequence, and many subtypes are further differentiated into toxin variants. Previous work in our laboratory described the use of a proteomics approach to distinguish subtype BoNT/A1 from BoNT/A2 where BoNT identities were confirmed after searching data against a database containing protein sequences of all known BoNT/A subtypes. We now describe here a similar approach to differentiate subtypes BoNT/B1, /B2, /B3, /B4, and /B5. Additionally, to identify new subtypes or hitherto unpublished amino acid substitutions, we created an amino acid substitution database covering every possible amino acid change. We used this database to differentiate multiple toxin variants within subtypes of BoNT/B1 and B2. More importantly, with our amino acid substitution database, we were able to identify a novel BoNT/B subtype, designated here as BoNT/B7. These techniques allow for subtype and strain level identification of both known and unknown BoNT/B rapidly with no DNA required. Figure Identification of an existing or new BoNT/B can be accomplished through MS/MS analysis of digestion fragments of the protein. Content Type Journal Article Category Original Paper Pages 1-12 DOI 10.1007/s00216-012-5767-3 Authors Suzanne R. Kalb, Centers for Disease Control and Prevention, National Center for Environmental Health, Division of Laboratory Sciences, 4770 Buford Hwy, N.E., Atlanta, GA 30341, USA Jakub Baudys, Centers for Disease Control and Prevention, National Center for Environmental Health, Division of Laboratory Sciences, 4770 Buford Hwy, N.E., Atlanta, GA 30341, USA Jon C. Rees, Centers for Disease Control and Prevention, National Center for Environmental Health, Division of Laboratory Sciences, 4770 Buford Hwy, N.E., Atlanta, GA 30341, USA Theresa J. Smith, Integrated Toxicology, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, MD 21702, USA Leonard A. Smith, Office of the Chief Scientist, Medical Research and Materiel Command (MRMC), Fort Detrick, MD 21702, USA Charles H. Helma, Bioscience Division Los Alamos National Laboratory, Los Alamos, NM 87545, USA Karen Hill, Bioscience Division Los Alamos National Laboratory, Los Alamos, NM 87545, USA Skadi Kull, Robert Koch-Institut, Center for Biological Security, Microbial Toxins (ZBS3), 13353 Berlin, Germany Sebastian Kirchner, Robert Koch-Institut, Center for Biological Security, Microbial Toxins (ZBS3), 13353 Berlin, Germany Martin B. Dorner, Robert Koch-Institut, Center for Biological Security, Microbial Toxins (ZBS3), 13353 Berlin, Germany Brigitte G. Dorner, Robert Koch-Institut, Center for Biological Security, Microbial Toxins (ZBS3), 13353 Berlin, Germany James L. Pirkle, Centers for Disease Control and Prevention, National Center for Environmental Health, Division of Laboratory Sciences, 4770 Buford Hwy, N.E., Atlanta, GA 30341, USA John R. Barr, Centers for Disease Control and Prevention, National Center for Environmental Health, Division of Laboratory Sciences, 4770 Buford Hwy, N.E., Atlanta, GA 30341, USA Journal Analytical and Bioanalytical Chemistry Online ISSN 1618-2650 Print ISSN 1618-2642
Print ISSN:
1618-2642
Topics:
Chemistry and Pharmacology
Permalink