Discovery of a novel enzymatic cleavage site for botulinum neurotoxin F5.

Centers for Disease Control and Prevention, National Center for Environmental Health, Division of Laboratory Sciences, 4770 Buford Hwy, NE, Atlanta, GA 30341, USA.
FEBS letters (Impact Factor: 3.54). 12/2011; 586(2):109-15. DOI: 10.1016/j.febslet.2011.11.033
Source: PubMed

ABSTRACT Botulinum neurotoxins (BoNTs) cause botulism by cleaving proteins necessary for nerve transmission. There are seven serotypes of BoNT, A-G, characterized by their response to antisera. Many serotypes are further distinguished into differing subtypes based on amino acid sequence, some of which result in functional differences. Our laboratory previously reported that all tested subtypes within each serotype have the same site of enzymatic activity. Recently, three new subtypes of BoNT/F; /F3, /F4, and /F5, were reported. Here, we report that BoNT/F5 cleaves substrate synaptobrevin-2 in a different location than the other BoNT/F subtypes, between (54)L and (55)E. This is the first report of cleavage of synaptobrevin-2 in this location.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Botulinum neurotoxin (BoNT) causes the disease known as botulism, which can be lethal. Rapid determination of exposure to BoNT is an important public health goal. Our laboratory has developed Endopep-MS, a mass spectrometry-based endopeptidase method for detecting and differentiating BoNT. Here, we demonstrate that this method is very sensitive, detecting as little as 0.5 mouse LD<sub align="right"> 50 </sub> of BoNT/A and as little as 0.05 mouse LD<sub align="right"> 50 </sub> of BoNT/B, /E, and /F spiked into human serum samples. Additionally, the ability to further differentiate BoNT as the subtype of BoNT/A spiked into milk using toxin proteomics and mass spectrometry has been demonstrated. This method does not require DNA and can be performed on the same sample as that used for Endopep-MS analysis. The combination of these techniques, all performed on the same sample, provides a sensitive and selective analysis of BoNT isolated from a food or clinical sample and measures the toxin's activity.
    The Botulinum J 01/2012; 2(2):119 - 134.
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Botulinum neurotoxins (BoNTs) are produced by anaerobic bacteria of the genus Clostridium and cause a persistent paralysis of peripheral nerve terminals, which is known as botulism. Neurotoxigenic clostridia belong to six phylogenetically distinct groups and produce more than 40 different BoNT types, which inactivate neurotransmitter release owing to their metalloprotease activity. In this Review, we discuss recent studies that have improved our understanding of the genetics and structure of BoNT complexes. We also describe recent insights into the mechanisms of BoNT entry into the general circulation, neuronal binding, membrane translocation and neuroparalysis.
    Nature Reviews Microbiology 06/2014; · 23.32 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Botulinum neurotoxin type F (BoNT/F) may be produced by Clostridium botulinum alone or in combination with another toxin type such as types A or B. Type F neurotoxin gene sequences have been further classified into 7 toxin subtypes. Recently, the genome sequence of one strain of C. botulinum (Af84) was shown to contain three neurotoxin genes (bont/F4, bont/F5, and bont/A2). In this study, a total of 8 strains containing bont/F4 and 7 strains containing bont/F5 were examined. Culture supernatants produced by these strains were incubated with BoNT/F-specific peptide substrates. Cleavage products of these peptides were subjected to mass spectral analysis allowing detection of the BoNT/F subtypes present in the culture supernatants. PCR analysis demonstrated that a plasmid-specific marker (PL-6) was observed only among strains containing bont/F5. Among these strains, Southern hybridization revealed the presence of an approximately 242-kb plasmid harboring bont/F5-. Genome sequencing of four of these strains revealed that the genomic backgrounds of strains harboring either bont/F4 or bont/F5 are diverse. None of the strains analyzed in this study were shown to produce BoNT/F4 and BoNT/F5 simultaneously suggesting that strain Af84 is unusual. Finally, these data support a role for the mobility of a bont/F5-encoding plasmid among strains of diverse genomic backgrounds.
    Applied and Environmental Microbiology 03/2014; · 3.95 Impact Factor

Full-text (2 Sources)

Available from
May 21, 2014