dc.contributor.author | Wang, Jiafu | |
dc.contributor.author | Zurawski, Tomas H. | |
dc.contributor.author | Meng, Jianghui | |
dc.contributor.author | Lawrence, Gary | |
dc.contributor.author | Olango, Weredeselam M. | |
dc.contributor.author | Finn, David P. | |
dc.contributor.author | Wheeler, Larry | |
dc.contributor.author | Dolly, J. Oliver | |
dc.date.accessioned | 2018-09-20T16:28:05Z | |
dc.date.available | 2018-09-20T16:28:05Z | |
dc.date.issued | 2010-12-07 | |
dc.identifier.citation | Wang, Jiafu; Zurawski, Tomas H. Meng, Jianghui; Lawrence, Gary; Olango, Weredeselam M.; Finn, David P.; Wheeler, Larry; Dolly, J. Oliver (2010). A dileucine in the protease of botulinum toxin a underlies its long-lived neuroparalysis. Journal of Biological Chemistry 286 (8), 6375-6385 | |
dc.identifier.issn | 0021-9258,1083-351X | |
dc.identifier.uri | http://hdl.handle.net/10379/14366 | |
dc.description.abstract | Blockade of neurotransmitter release by botulinum neurotoxin type A (BoNT(A)) underlies the severe neuroparalytic symptoms of human botulism, which can last a few years. The structural basis for this remarkable persistence remains unclear. Herein, recombinant BoNT(A) was found to match the neurotoxicity of that from Clostridium botulinum, producing persistent cleavage of synaptosomal-associated protein of 25 kDa (SNAP-25) and neuromuscular paralysis. When two leucines near the C terminus of the protease light chain of A (LC(A)) were mutated, its inhibition of exocytosis was followed by fast recovery of intact SNAP-25 in cerebellar neurons and neuromuscular transmission in vivo. Deletion of 6-7 N terminus residues diminished BoNT(A) activity but did not alter the longevity of its SNAP-25 cleavage and neuromuscular paralysis. Furthermore, genetically fusing LC(E) to a BoNT(A) enzymically inactive mutant (BoTIM(A)) yielded a novel LC(E)-BoTIM(A) protein that targets neurons, and the BoTIM(A) moiety also delivers and stabilizes the inhibitory LC(E), giving a potent and persistent cleavage of SNAP-25 with associated neuromuscular paralysis. Moreover, its neurotropism was extended to sensory neurons normally insensitive to BoNT(E). LC(E)-BoTIM(A)(AA) with the above-identified dileucine mutated gave transient neuromuscular paralysis similar to BoNT(E), reaffirming that these residues are critical for the persistent action of LC(E)-BoTIM(A) as well as BoNT(A). LC(E)-BoTIM(A) inhibited release of calcitonin gene-related peptide from sensory neurons mediated by transient receptor potential vanilloid type 1 and attenuated capsaicin-evoked nociceptive behavior in rats, following intraplantar injection. Thus, a long acting, versatile composite toxin has been developed with therapeutic potential for pain and conditions caused by overactive cholinergic nerves. | |
dc.publisher | American Society for Biochemistry & Molecular Biology (ASBMB) | |
dc.relation.ispartof | Journal of Biological Chemistry | |
dc.rights | Attribution-NonCommercial-NoDerivs 3.0 Ireland | |
dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/3.0/ie/ | |
dc.subject | neurotoxin-e | |
dc.subject | neuromuscular-transmission | |
dc.subject | nerve-terminals | |
dc.subject | synaptotagmin-i | |
dc.subject | sensory neurons | |
dc.subject | translocation | |
dc.subject | exocytosis | |
dc.subject | persistence | |
dc.subject | inhibition | |
dc.subject | injection | |
dc.title | A dileucine in the protease of botulinum toxin a underlies its long-lived neuroparalysis | |
dc.type | Article | |
dc.identifier.doi | 10.1074/jbc.m110.181784 | |
dc.local.publishedsource | http://www.jbc.org/content/286/8/6375.full.pdf | |
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