Studies on botulinum neurotoxins type /C1 and mosaic/DC using Endopep-MS and proteomics

Hercules Moura, Rebecca R. Terilli, Adrian R. Woolfitt, Maribel Gallegos-Candela, Lisa G. Mcwilliams, Maria I. Solano, James L. Pirkle, John R. Barr

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

Botulinum neurotoxins (BoNTs) are very potent toxins and category A biological threat agents. BoNT serotypes /C1 and /D affect birds and mammals and can be potentially lethal to humans. We have previously described the usefulness of the Endopep-MS method to detect the activity of BoNT A through G. This report was followed by the application of the method to clinical samples. The activity of the BoNT serotypes associated with human disease (/A, /B, /E, and /F) was successfully detected. However, BoNT/C and /D require different conditions for fast substrate cleavage, and a comprehensive description of a method to study BoNT/C and /D has not yet been reported. This work describes a new, optimized version of the Endopep-MS method to detect BoNTs /C1 and /DC either spiked directly in 20μL of reaction buffer or spiked in a larger volume of buffer and further extracted using antibody-coated magnetic beads. It was found that the incubation temperature at 42°C was more effective for both toxin serotypes, but each toxin serotype has an optimum cleavage pH. Additionally, we describe for the first time a proteomics study using a fast trypsin digestion method and label-free quantification of these toxin serotypes. FEMS Immunology & Medical Microbiology

Original languageEnglish (US)
Pages (from-to)288-300
Number of pages13
JournalFEMS Immunology and Medical Microbiology
Volume61
Issue number3
DOIs
StatePublished - Apr 2011
Externally publishedYes

Keywords

  • Botulism
  • Endopep-MS
  • Label-free quantification
  • MS
  • Proteomics

ASJC Scopus subject areas

  • Immunology and Allergy
  • Microbiology
  • Immunology
  • Microbiology (medical)
  • Infectious Diseases

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