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Recombinant botulinum neurotoxin A heavy chain-based delivery vehicles for neuronal cell targeting.
- Source :
-
Protein engineering, design & selection : PEDS [Protein Eng Des Sel] 2011 Mar; Vol. 24 (3), pp. 247-53. Date of Electronic Publication: 2010 Nov 04. - Publication Year :
- 2011
-
Abstract
- The long half-life of botulinum neurotoxin serotype A (BoNT/A) in cells poses a challenge in developing post-exposure therapeutics complementary to existing antitoxin strategies. Delivery vehicles consisting of the toxin heavy chain (HC), including the receptor-binding domain and translocation domain, connected to an inhibitory cargo offer a possible solution for rescuing intoxicated neurons in victims paralyzed from botulism. Here, we report the expression and purification of soluble recombinant prototype green fluorescent protein (GFP) cargo proteins fused to the entire BoNT/A-HC (residues 544-1295) in Escherichia coli with up to a 40 amino acid linker inserted between the cargo and BoNT/A-HC vehicle. We show that these GFP-HC fusion proteins are functionally active and readily taken up by cultured neuronal cells as well as by neuronal cells in mouse motor nerve endings.
- Subjects :
- Animals
Botulinum Toxins, Type A isolation & purification
Botulinum Toxins, Type A metabolism
Drug Carriers chemistry
Drug Carriers isolation & purification
Drug Carriers metabolism
Escherichia coli genetics
Mice
Motor Neurons cytology
Motor Neurons metabolism
Neuromuscular Junction cytology
Protein Engineering
Protein Structure, Tertiary
Protein Transport
Recombinant Fusion Proteins isolation & purification
Recombinant Fusion Proteins metabolism
Reflex drug effects
Toes physiology
Botulinum Toxins, Type A chemistry
Botulinum Toxins, Type A genetics
Drug Delivery Systems methods
Neurons cytology
Neurons metabolism
Recombinant Fusion Proteins chemistry
Recombinant Fusion Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1741-0134
- Volume :
- 24
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Protein engineering, design & selection : PEDS
- Publication Type :
- Academic Journal
- Accession number :
- 21051321
- Full Text :
- https://doi.org/10.1093/protein/gzq093