Engineering botulinum neurotoxin domains for activation by toxin light chain.

摘要: Targeted secretion inhibitors (TSI) are a new class of biopharmaceuticals designed from botulinum neurotoxin protein scaffold. The backbone consists the 50-kDa endopeptidase light chain and translocation domain (N-terminal portion heavy chain), lacks neuronal toxicity, but retains ability to target cytoplasmic soluble N-ethylmaleimide-sensitive factor attachment receptor (SNARE) proteins. TSI produced as single-chain proteins then cleaved post-translationally generate functional heterodimers. Precise proteolytic cleavage is essential activate dichain form. themselves highly specific proteases. We have exploited this activity create self-activating enzymes by replacing native site with substrate SNARE peptide for protease. also created cross-activating backbones. By activation in one another serotype, controlled achieved. peptides encompassing whole coiled-coil region enabled complete assembly backbone. These engineered backbones capable translocating their enzymatic domains intracellular They investigative tools which further understanding interactions.