title: Synthetic self-assembling clostridial chimera for modulation of sensory functions.
abstract: Clostridial neurotoxins reversibly block neuronal communication for weeks and months. While these proteolytic neurotoxins hold great promise for clinical applications and the investigation of brain function, their paralytic activity at neuromuscular junctions is a stumbling block. To redirect the clostridial activity to neuronal populations other than motor neurons, we used a new self-assembling method to combine the botulinum type A protease with the tetanus binding domain, which natively targets central neurons. The two parts were produced separately and then assembled in a site-specific way using a newly introduced 'protein stapling' technology. Atomic force microscopy imaging revealed dumbbell shaped particles which measure ∼23 nm. The stapled chimera inhibited mechanical hypersensitivity in a rat model of inflammatory pain without causing either flaccid or spastic paralysis. Moreover, the synthetic clostridial molecule was able to block neuronal activity in a defined area of visual cortex. Overall, we provide the first evidence that the protein stapling technology allows assembly of distinct proteins yielding new biomedical properties.
authors.affiliation: MRC Laboratory of Molecular Biology , Cambridge, United Kingdom.
authors.forename: Enrico
authors.initials: E
authors.lastname: Ferrari
authors.forename: Chunjing
authors.initials: C
authors.lastname: Gu
authors.forename: Dhevahi
authors.initials: D
authors.lastname: Niranjan
authors.forename: Laura
authors.initials: L
authors.lastname: Restani
authors.forename: Christine
authors.initials: C
authors.lastname: Rasetti-Escargueil
authors.forename: Ilona
authors.initials: I
authors.lastname: Obara
authors.forename: Sandrine M
authors.initials: SM
authors.lastname: Geranton
authors.forename: Jason
authors.initials: J
authors.lastname: Arsenault
authors.forename: Tom A
authors.initials: TA
authors.lastname: Goetze
authors.forename: Callista B
authors.initials: CB
authors.lastname: Harper
authors.forename: Tam H
authors.initials: TH
authors.lastname: Nguyen
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Maywood
authors.forename: John
authors.initials: J
authors.lastname: O'Brien
authors.forename: Giampietro
authors.initials: G
authors.lastname: Schiavo
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Wheeler
authors.forename: Frederic A
authors.initials: FA
authors.lastname: Meunier
authors.forename: Michael
authors.initials: M
authors.lastname: Hastings
authors.forename: J Michael
authors.initials: JM
authors.lastname: Edwardson
authors.forename: Dorothea
authors.initials: D
authors.lastname: Sesardic
authors.forename: Matteo
authors.initials: M
authors.lastname: Caleo
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Hunt
authors.forename: Bazbek
authors.initials: B
authors.lastname: Davletov
mesh_terms: D000818:Animals; D019274:Botulinum Toxins, Type A / Q000008:administration & dosage / Q000378:metabolism*; D001921:Brain / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D003014:Clostridium botulinum / Q000378:metabolism; D003017:Clostridium tetani / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D009474:Neurons / Q000166:cytology / Q000187:drug effects; D017288:Pain Threshold / Q000187:drug effects*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000378:metabolism*; D013744:Tetanus Toxin / Q000008:administration & dosage / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D013744:Tetanus Toxin; D019274:Botulinum Toxins, Type A