From the Mouse to the Mass Spectrometer: Detection and Differentiation of the Endoproteinase Activities of Botulinum Neurotoxins A−G by Mass Spectrometry
2005; American Chemical Society; Volume: 77; Issue: 13 Linguagem: Inglês
10.1021/ac050485f
ISSN1520-6882
AutoresAnne E. Boyer, Hércules Moura, Adrian R. Woolfitt, Suzanne R. Kalb, Lisa G. McWilliams, Antonis J. Pavlopoulos, Jürgen Schmidt, David L. Ashley, John R. Barr,
Tópico(s)Biochemical and Structural Characterization
ResumoWe have developed an assay (Endopep-MS) that detects the specific endoproteinase activities of all seven BoNT types by mass spectrometry (MS). Each BoNT type cleaves a unique site on proteins involved in neuronal transmission. Target peptide substrates based on these proteins identify a BoNT type by its enzymatic action on the substrate and the production of two peptide products, which are then detected by matrix-assisted laser desorption/ionization time-of-flight MS or liquid chromatography electrospray ionization MS/MS. We showed the ability to detect all seven toxin types in a multiplexed assay format. The detection limits achieved range from 0.039 to 0.625 mouse LD(50)/mL for toxin types A, B, E, and F in a buffer system. The Endopep-MS assay is the first to differentiate all seven BoNT types, is sensitive, specific, and has the potential to quantify toxin activity.
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