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Characterisation of botulinum toxins type A and B, by matrix-assisted laser desorption ionisation and electrospray mass spectrometry

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Author: Baar, B.L.M. van · Hulst, A.G. · Jong, A.L. de · Wils, E.R.J.
Type:article
Date:2002
Institution: Prins Maurits Laboratorium TNO
Source:Journal of Chromatography A, 1-2, 970, 95-115
Identifier: 236690
doi: doi:10.1016/S0021-9673(02)00508-3
Keywords: Amino acid sequencing · Botulinum toxins · Mass spectrometry · Peptide mapping · Toxins · Amino acids · Bacteria · Desorption · Ionization · Laser applications · Mass spectrometry · Military operations · Proteins · Electrospray · Toxic materials · Botulinum toxin A · Botulinum toxin B · Hemagglutinin · Pepsin A · Trypsin · Amino acid sequence · Analytic method · Article · Bacterial strain · Clostridium botulinum · Data base · Gene sequence · Liquid chromatography · Mass spectrometry · Priority journal · Protein analysis · Sequence alignment · Amino Acid Sequence · Botulinum Toxin Type A · Botulinum Toxins · Molecular Sequence Data · Pepsin A · Sequence Homology, Amino Acid · Spectrometry, Mass, Electrospray Ionization · Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization · Trypsin · Abelmoschus · Bacteria (microorganisms) · Clostridium · Clostridium botulinum

Abstract

A method earlier developed for the mass spectrometric (MS) identification of tetanus toxin (TTx) was applied to botulinum toxins type A and B (BTxA and BTxB). Botulinum toxins are extremely neurotoxic bacterial toxins, likely to be used as biological warfare agent. Biologically active BTxA and BTxB are comprised of a protein complex of the respective neurotoxins with specific haemagglutinins (HAs) and non-toxic non-haemagglutinins (NTNHs). These protein complexes are also observed in mass spectrometric identification. The particular BTxA complex, from Clostridium botulinum strain 62A, almost completely matched database data derived from genetic sequences known for this strain. Although no such database information was available for BTxB, from C. botulinum strain okra, all protein sequences from the complex except that of HA-70 were found to match proteins known from other type B strains. It was found that matrix-assisted laser desorption ionisation MS provides provisional identification from trypsin digest peptide maps and that liquid chromatography electrospray (tandem) mass spectrometry affords unequivocal identification from amino acid sequence information of digest peptides obtained in trypsin or pepsin digestion. © 2002 Elsevier Science B.V. All rights reserved. Molecular Sequence Numbers: D67030, L42537, M30196, M81186, U24431, U63808, X52066, X79102, X79104, X92973, Y13630, Y14239; Chemicals/CAS: Botulinum Toxin Type A; botulinum toxin type B; Botulinum Toxins; Pepsin A, EC 3.4.23.1; Trypsin, EC 3.4.21.4