A yeast assay probes the interaction between botulinum neurotoxin serotype B and its SNARE substrate.

Proceedings of the National Academy of Sciences of the United States of America
Hong FangNeil Green

Abstract

The seven functionally distinct serotypes (A-G) of botulinum neurotoxin (BoNT) are dichains consisting of light chain (LC) with zinc-dependent endoprotease activity connected by one disulfide bond to heavy chain with neuronal-cell translocation and receptor-binding domains. LC-mediated proteolysis of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins and consequent inhibition of synaptic vesicle fusion to the presynaptic membrane of human motor neurons are responsible for flaccid paralysis associated with botulism. LC endoproteolysis is complex, requiring highly extended SNARE sequences at the surface of intracellular membranes and prompting our development of a genetically amenable assay to monitor the interaction between BoNT/LC and its SNARE substrate. Using BoNT serotype B as a model, the assay employs a chimeric SNARE protein where a portion of neuronal synaptobrevin (Sb) is fused to Snc2p, a Sb ortholog required for protein secretion from yeast cells. Regulated expression of serotype B-LC in yeast leads to cleavage of the chimera and a conditional growth defect. To assess utility of this assay for monitoring SNARE protein cleavage, we growth-selected chimeric SNARE mutations that inhibi...Continue Reading

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Citations

Mar 5, 2009·Biochemistry·James S HenkelJoseph T Barbieri
Jun 12, 2013·Cellular and Molecular Life Sciences : CMLS·Sergio Pantano, Cesare Montecucco
Mar 6, 2010·Trends in Microbiology·Ramin Mollaaghababa HakamiSina Bavari
Apr 3, 2012·Toxicon : Official Journal of the International Society on Toxinology·Mojca MattiazziIgor Križaj
Nov 10, 2011·Toxins·Andy Pickett, Karen Perrow

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Botulism is a rare but serious paralytic illness caused by a nerve toxin that is produced by the bacterium clostridium botulinum. Discover the latest research on botulism here.

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Botulism is a rare but serious paralytic illness caused by a nerve toxin that is produced by the bacterium clostridium botulinum. Discover the latest research on botulism here.