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 2.8Å 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.
|
Authors | Geoffrey Masuyer, Matthew Beard, Verity A Cadd, John A Chaddock, K Ravi Acharya |
Journal | Journal of structural biology
(J Struct Biol)
Vol. 174
Issue 1
Pg. 52-7
(Apr 2011)
ISSN: 1095-8657 [Electronic] United States |
PMID | 21078393
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
|
Copyright | Copyright © 2010 Elsevier Inc. All rights reserved. |
Chemical References |
- Vesicle-Associated Membrane Protein 1
- Vesicle-Associated Membrane Protein 2
- rimabotulinumtoxinB
- Botulinum Toxins
- Botulinum Toxins, Type A
|
Topics |
- Amino Acid Sequence
- Animals
- Botulinum Toxins
(chemistry, genetics, metabolism)
- Botulinum Toxins, Type A
- Cells, Cultured
- Clostridium botulinum
(metabolism)
- Crystallography, X-Ray
- Humans
- Molecular Sequence Data
- Protein Structure, Secondary
- Rats
- Rats, Sprague-Dawley
- Sequence Homology, Amino Acid
- Structure-Activity Relationship
- Vesicle-Associated Membrane Protein 1
(metabolism)
- Vesicle-Associated Membrane Protein 2
(metabolism)
|