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Structure and activity of a functional derivative of Clostridium botulinum neurotoxin

DOI: 10.1016/j.jsb.2010.11.010 DOI Help
PMID: 21078393 PMID Help

Authors: Geoffrey Masuyer (University of Bath) , Matthew Beard (Syntaxin Limited) , Verity Cadd (Syntaxin Limited) , John Chaddock (Syntaxin Limited) , K. Ravi Acharya (University of Bath)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Journal Of Structural Biology , VOL 174 (1) , PAGES 52-57

State: Published (Approved)
Published: April 2011

Abstract: Botulinum neurotoxins (BoNTs) cause flaccid paralysis by inhibiting neurotransmission at cholinergic nerve terminals. BoNTs consist of three essential domains for toxicity: the cell binding domain (Hc), the translocation domain (Hn) and the catalytic domain (LC). A functional derivative (LHn) of the parent neurotoxin B composed of Hn and LC domains was recombinantly produced and characterised. LHn/B crystallographic structure at 2.8 Å resolution is reported. The catalytic activity of LHn/B towards recombinant human VAMP was analysed by substrate cleavage assay and showed a higher specificity for VAMP-1, -2 compared to VAMP-3. LHn/B also showed measurable activity in living spinal cord neurons. Despite lacking the Hc (cell-targeting) domain, LHn/B retained the capacity to internalize and cleave intracellular VAMP-1 and -2 when added to the cells at high concentration. These activities of the LHn/B fragment demonstrate the utility of engineered botulinum neurotoxin fragments as analytical tools to study the mechanisms of action of BoNT neurotoxins and of SNARE proteins.

Journal Keywords: Animals; Botulinum; Cells; Cultured; Clostridium; Crystallography; X-Ray; Humans; Molecular; Protein; Secondary; Rats; Rats; Sprague-Dawley; Sequence; Amino; Structure-Activity; Vesicle-Associated; Vesicle-Associated Membrane Protein 2

Subject Areas: Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography

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