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

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

Research output: Contribution to journalArticle

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 1 2011
Externally publishedYes

Fingerprint

Neurotoxins
Proteomics
Buffers
Type A Botulinum Toxins
Biological Factors
Microbiology
Allergy and Immunology
Trypsin
Birds
Digestion
Mammals
Serogroup
Temperature
Antibodies
botulinum toxin type C
botulinum toxin type D

Keywords

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

ASJC Scopus subject areas

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

Cite this

Studies on botulinum neurotoxins type /C1 and mosaic/DC using Endopep-MS and proteomics. / Moura, Hercules; Terilli Veenhuis, Rebecca; Woolfitt, Adrian R.; Gallegos-Candela, Maribel; Mcwilliams, Lisa G.; Solano, Maria I.; Pirkle, James L.; Barr, John R.

In: FEMS Immunology and Medical Microbiology, Vol. 61, No. 3, 01.04.2011, p. 288-300.

Research output: Contribution to journalArticle

Moura, H, Terilli Veenhuis, R, Woolfitt, AR, Gallegos-Candela, M, Mcwilliams, LG, Solano, MI, Pirkle, JL & Barr, JR 2011, 'Studies on botulinum neurotoxins type /C1 and mosaic/DC using Endopep-MS and proteomics', FEMS Immunology and Medical Microbiology, vol. 61, no. 3, pp. 288-300. https://doi.org/10.1111/j.1574-695X.2010.00774.x
Moura, Hercules ; Terilli Veenhuis, Rebecca ; Woolfitt, Adrian R. ; Gallegos-Candela, Maribel ; Mcwilliams, Lisa G. ; Solano, Maria I. ; Pirkle, James L. ; Barr, John R. / Studies on botulinum neurotoxins type /C1 and mosaic/DC using Endopep-MS and proteomics. In: FEMS Immunology and Medical Microbiology. 2011 ; Vol. 61, No. 3. pp. 288-300.
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