Recombinant derivatives of botulinum neurotoxins engineered for trafficking studies and neuronal delivery

摘要

This invention relates to isolated Clostridium botulinum propeptides and neurotoxins, isolated nucleic acid molecules encoding Clostridium botulinum propeptides and neurotoxins, methods of expression, treatment methods, and methods of detecting neurotoxin trafficking. The isolated Clostridium botulinum propeptides have a light chain region; a heavy chain region, where the light and heavy chain regions are linked by a disulfide bond; an intermediate region connecting the light and heavy chain regions and comprising a highly specific protease cleavage site; and an S6 peptide sequence according to SEQ ID NO:2 positioned upstream from, but not attached directly to, the N-terminus of the neurotoxin propeptide at the light chain region to enable site specific attachment of cargo.

主权项

1. An isolated Clostridium botulinum neurotoxin propeptide comprising:
a light chain region; a heavy chain region, wherein the light and heavy chain regions are linked by a disulfide bond; an intermediate region connecting the light and heavy chain regions and comprising a highly specific protease cleavage site, wherein said highly specific protease cleavage site has three or more specific adjacent amino acid residues that are recognized by the highly specific protease in order to enable cleavage; and a cargo attachment peptide sequence to enable site-specific attachment of cargo, wherein the cargo attachment peptide sequence is positioned upstream of the light chain region and is separated from the N-terminus of the light chain region by an amino acid spacer sequence.