Print Email Facebook Twitter Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry Title Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry Author van Baar, B.L.M. Hulst, A.G. de Jong, A.L. Wils, E.R.J. Prins Maurits Laboratorium TNO Publication year 2004 Abstract In a follow-up of the earlier characterisation of botulinum toxins type A and B (BTxA and BTxB) by mass spectrometry (MS), types C, D, E, and F (BTxC, BTxD, BTxE, BTxF) were now investigated. Botulinum toxins are extremely neurotoxic bacterial toxins, likely to be used as biological warfare agent. Biologically active BTxC, BTxD, BTxE, and BTxF are comprised of a protein complex of the respective neurotoxins with non-toxic non-haemagglutinin (NTNH) and, sometimes, specific haemagglutinins (HA). These protein complexes were observed in mass spectrometric identification. The BTxC complex, from Clostridium botulinum strain 003-9, consisted of a 'type C1 and D mosaic' toxin similar to that of type C strain 6813, a non-toxic non-hemagglutinating and a 33kDa hemagglutinating (HA-33) component similar to those of strain C-Stockholm, and an exoenzyme C3 of which the sequence was in full agreement with the known genetic sequence of strain 003-9. The BTxD complex, from C. botulinum strain CB-16, consisted of a neurotoxin with the observed sequence identical with that of type D strain BVD/-3 and of an NTNH with the observed sequence identical with that of type C strain C-Yoichi. Remarkably, the observed protein sequence of CB-16 NTNH differed by one amino acid from the known gene sequence: L859 instead of F859. The BTxE complex, from a C. botulinum isolated from herring sprats, consisted of the neurotoxin with an observed sequence identical with that from strain NCTC 11219 and an NTNH similar to that from type E strain Mashike (1 amino acid difference with observed sequence). BTxF, from C. botulinum strain Langeland (NCTC 10281), consisted of the neurotoxin and an NTNH; observed sequences from both proteins were in agreement with the gene sequence known from strain Langeland. As with BTxA and BTxB, matrix-assisted laser desorption/ionisation (MALDI) MS provided provisional identification from trypsin digest peptide maps and liquid chromatography-electrospray (tandem) mass spectrometry (LC-ES MS) afforded unequivocal identification from amino acid sequence information of digest peptides obtained in trypsin digestion. © 2004 Elsevier B.V. All rights reserved. Chemicals / CAS: hemagglutinin, 37333-12-3; neurotoxin, 39386-17-9; trypsin, 9002-07-7; Botulinum Toxins Subject Amino acid sequencingBotulinum toxinsMass spectrometryPeptide mappingToxinsGenesIonizationLaser applicationsLiquid chromatographyMass spectrometryProteinsBotulinum toxinsGene sequencesBacteriaBotulinum toxinBotulinum toxin cBotulinum toxin dBotulinum toxin EBotulinum toxin fHemagglutininNeurotoxinTrypsinUnclassified drugAmino acid sequenceAnalytic methodArticleClostridium botulinumControlled studyElectrospray mass spectrometryGene sequenceHemagglutinationHerringLiquid chromatographyMatrix assisted laser desorption ionization time of flight mass spectrometryNonhumanPriority journalTandem mass spectrometryAmino Acid SequenceBotulinum ToxinsMolecular Sequence DataPeptide MappingSpectrometry, Mass, Electrospray IonizationSpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationBacteria (microorganisms)ClostridiumClostridium botulinumClupea harengusClupeidae To reference this document use: http://resolver.tudelft.nl/uuid:6557f75d-e764-4697-bcb9-772ebf171df4 DOI https://doi.org/10.1016/j.chroma.2004.02.047 TNO identifier 237719 ISSN 0021-9673 Source Journal of Chromatography A, 1035 (1), 97-114 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.