Light Chain of Botulinum A Neurotoxin Expressed as an Inclusion Body from a Synthetic Gene is Catalytically and Functionally Active
Abstract
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 (BoNT/A), 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.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2000
- Accession Number
- ADA386484
Entities
People
- Leonard A. Smith
- S. A. Ahmed
Organizations
- United States Army Medical Research Institute of Infectious Diseases