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SNARE tagging allows stepwise assembly of a multimodular medicinal toxin

Darios, Frédéric, Niranjan, Dhevahi, Ferran, Enrico, Zhang, Fan, Soloviev, Mikhail, Rummel, Andreas, Bigalke, Hans, Suckling, Jason, Ushkaryov, Yuri, Naumenko, Nikolay, and others. (2010) SNARE tagging allows stepwise assembly of a multimodular medicinal toxin. Proceedings of the National Academy of Sciences, 107 (42). pp. 18197-18201. ISSN 1091-6490. (doi:10.1073/pnas.1007125107) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided. (Contact us about this Publication)
Official URL
http://dx.doi.org/10.1073/pnas.1007125107

Abstract

Generation of supramolecular architectures through controlled linking of suitable building blocks can offer new perspectives to medicine and applied technologies. Current linking strategies often rely on chemical methods that have limitations and cannot take full advantage of the recombinant technologies. Here we used SNARE proteins, namely, syntaxin, SNAP25, and synaptobrevin, which form stable tetrahelical complexes that drive fusion of intracellular membranes, as versatile tags for irreversible linking of recombinant and synthetic functional units. We show that SNARE tagging allows stepwise production of a functional modular medicinal toxin, namely, botulinum neurotoxin type A, commonly known as BOTOX. This toxin consists of three structurally independent units: Receptor-binding domain (Rbd), Translocation domain (Td), and the Light chain (Lc), the last being a proteolytic enzyme. Fusing the receptor-binding domain with synaptobrevin SNARE motif allowed delivery of the active part of botulinum neurotoxin (Lc-Td), tagged with SNAP25, into neurons. Our data show that SNARE-tagged toxin was able to cleave its intraneuronal molecular target and to inhibit release of neurotransmitters. The reassembled toxin provides a safer alternative to existing botulinum neurotoxin and may offer wider use of this popular research and medical tool. Finally, SNARE tagging allowed the Rbd portion of the toxin to be used to deliver quantum dots and other fluorescent markers into neurons, showing versatility of this unique tagging and self-assembly technique. Together, these results demonstrate that the SNARE tetrahelical coiled-coil allows controlled linking of various building blocks into multifunctional assemblies.

Item Type: Article
DOI/Identification number: 10.1073/pnas.1007125107
Additional information: The author made a substantial contribution to the organisation of the conduct of the study, to carrying out the study (including acquisition of study data) and to analysis and interpretation of the study data and b) The author helped draft the output.; number of additional authors: 15;
Uncontrolled keywords: botulinum neurotoxin, protein linking, recominant, self-assembly, coiled coil.
Subjects: Q Science
R Medicine > RS Pharmacy and materia medica
Divisions: Faculties > Sciences > Medway School of Pharmacy
Depositing User: Stewart Brownrigg
Date Deposited: 07 Mar 2014 00:05 UTC
Last Modified: 29 May 2019 12:24 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/40390 (The current URI for this page, for reference purposes)
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