Targeted delivery of a SNARE protease to sensory neurons using a single chain antibody (scFv) against the extracellular domain of P2X(3) inhibits the release of a pain mediator.

Abstract	P2X3 (P2X purinoceptor 3) is predominantly expressed on nociceptive sensory neurons and plays a crucial role in signalling leading to chronic inflammatory pain and some features of neuropathic pain. Thus it represents a potential target for pain therapeutics. BoNT/A (botulinum neurooxin type A) effectively relieves certain types of pain through inhibiting the neuronal release of pain peptides. A recombinant single-chain variable fragment (scFv) antibody designated MH7C was generated against the extracellular domain of P2X3 using phage display. The genes encoding the scFv and activated di-chain form of BoNT/A without the C-terminal-binding subdomain (LC-HN-HCN/A) were ligated and expressed in Escherichia coli cells as a composite fusion protein. The purified protein bound and entered P2X3-containing sensory neurons, cleaved synaptosomal-associated protein of 25 kDa and inhibited the release of a pain peptide. This novel fusion protein designated 'LC-HN-HCN/A-MH7C' has potential clinical applications in the treatment of chronic inflammatory and sympathetically maintained neuropathic pain.
Authors	Hui Ma, Jianghui Meng, Jiafu Wang, Stephen Hearty, J Oliver Dolly, Richard O'Kennedy
Journal	The Biochemical journal (Biochem J) Vol. 462 Issue 2 Pg. 247-56 (Sep 01 2014) ISSN: 1470-8728 [Electronic] England
PMID	24844294 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References	Receptors, Purinergic P2X3 Recombinant Fusion Proteins Single-Chain Antibodies Synaptosomal-Associated Protein 25 Botulinum Toxins, Type A incobotulinumtoxinA Calcitonin Gene-Related Peptide
Topics	Animals Botulinum Toxins, Type A (chemistry, genetics) Calcitonin Gene-Related Peptide (metabolism) Cells, Cultured Female Ganglia, Spinal (cytology) Humans Mice Pain (metabolism) Rabbits Rats Receptors, Purinergic P2X3 (immunology) Recombinant Fusion Proteins (chemistry, genetics, pharmacology) Sensory Receptor Cells (drug effects, metabolism) Single-Chain Antibodies (chemistry, genetics) Synaptosomal-Associated Protein 25 (metabolism)

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