SNARE tagging allows stepwise assembly of a multimodular medicinal toxin

Darios, Frederic and Niranjan, Dhevahi and Ferrari, Enrico and Zhang, Fan and Soloviev, Mikhail and Rummel, Andreas and Bigalke, Hans and Suckling, Jason and Ushkaryov, Yuri and Naumenko, Nikolay and Shakirzyanova, Anastasia and Giniatullin, Rashid and Maywood, Elizabeth and Hastings, Michael and Binz, Thomas and Davletov, Bazbek (2010) SNARE tagging allows stepwise assembly of a multimodular medicinal toxin. Proceedings of the National Academy of Sciences, 107 (42). 18197-18201 . ISSN 1091-6490

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Full text 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
Keywords:botulinum neurotoxin, protein linking, recombinant, self-assembly, coiled coil
Subjects:B Subjects allied to Medicine > B140 Neuroscience
C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
Divisions:College of Science > School of Life Sciences
ID Code:6586
Deposited By: Enrico Ferrari
Deposited On:15 Oct 2012 20:44
Last Modified:29 Nov 2013 09:11

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