title: Re-engineering the target specificity of Clostridial neurotoxins - a route to novel therapeutics.
abstract: The ability to chemically couple proteins to LH(N)-fragments of clostridial neurotoxins and create novel molecules with selectivity for cells other than the natural target cell of the native neurotoxin is well established. Such molecules are able to inhibit exocytosis in the target cell and have the potential to be therapeutically beneficial where secretion from a particular cell plays a causative role in a disease or medical condition. To date, these molecules have been produced by chemical coupling of the LH(N)-fragment and the targeting ligand. This is, however, not a suitable basis for producing pharmaceutical agents as the products are ill defined, difficult to control and heterogeneous. Also, the molecules described to date have targeted neuroendocrine cells that are susceptible to native neurotoxins, and therefore the benefit of creating a molecule with a novel targeting domain has been limited. In this paper, the production of a fully recombinant fusion protein from a recombinant gene encoding both the LH(N)-domain of a clostridial neurotoxin and a specific targeting domain is described, together with the ability of such recombinant fusion proteins to inhibit secretion from non-neuronal target cells. Specifically, a novel protein consisting of the LH(N)-domains of botulinum neurotoxin type C and epidermal growth factor (EGF) that is able to inhibit secretion of mucus from epithelial cells is reported. Such a molecule has the potential to prevent mucus hypersecretion in asthma and chronic obstructive pulmonary disease.
authors.affiliation: Health Protection Agency, Centre for Emergency Preparedness & Response, Porton Down, Salisbury, UK. keith.foster@hpa.org.uk
authors.forename: K A
authors.initials: KA
authors.lastname: Foster
authors.forename: E J
authors.initials: EJ
authors.lastname: Adams
authors.forename: L
authors.initials: L
authors.lastname: Durose
authors.forename: C J
authors.initials: CJ
authors.lastname: Cruttwell
authors.forename: E
authors.initials: E
authors.lastname: Marks
authors.forename: C C
authors.initials: CC
authors.lastname: Shone
authors.forename: J A
authors.initials: JA
authors.lastname: Chaddock
authors.forename: C L
authors.initials: CL
authors.lastname: Cox
authors.forename: C
authors.initials: C
authors.lastname: Heaton
authors.forename: J M
authors.initials: JM
authors.lastname: Sutton
authors.forename: J
authors.initials: J
authors.lastname: Wayne
authors.forename: F C G
authors.initials: FC
authors.lastname: Alexander
authors.forename: D F
authors.initials: DF
authors.lastname: Rogers
mesh_terms: D001905:Botulinum Toxins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D002460:Cell Line; D010450:Endopeptidases / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D000922:Immunotoxins / Q000737:chemistry / Q000494:pharmacology; D037102:Lectins / Q000378:metabolism; D008024:Ligands; D009077:Mucins / Q000378:metabolism; D015202:Protein Engineering; D021381:Protein Transport; D050765:Qa-SNARE Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism
chemical_list: D000922:Immunotoxins; D037102:Lectins; D008024:Ligands; D009077:Mucins; D050765:Qa-SNARE Proteins; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D010450:Endopeptidases; D001905:Botulinum Toxins; C096324:botulinum toxin type C