Light Chain of Botulinum A Neurotoxin Expressed as an Inclusion Body from a Synthetic Gene is Catalytically and Functionally Active
ARMY MEDICAL RESEARCH INST OF INFECTIOUS DISEASES FORT DETRICK MD
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Botulinum neurotoxins, the most potent of all toxins, induce lethal neuromuscular paralysis by inhibiting exocytosis at the neuromuscular junction. The light chains LC of these dichain neurotoxins are a new class of zinc-endopeptidases that specifically cleave the synaptosomal proteins, SNAP-25, VAMP, or syntaxin at discrete sites. To facilitate the structural and functional characterization of these unique endopeptidases, we constructed a synthetic gene for the LC of the botulinum neurotoxin serotype A BoNTA, overexpressed it in Escherichia coli, and purified the gene product from inclusion bodies. Our procedure can provide 1.1g of the LC from 1 L of culture. The LC product was stable in solution at 4 C for at least 6 months.