Recombinant botulinum neurotoxin A heavy chain-based delivery vehicles for neuronal cell targeting.

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Title:	Recombinant botulinum neurotoxin A heavy chain-based delivery vehicles for neuronal cell targeting.
Authors:	Ho, Mengfei^1,2, Chang, Li-Hsin¹, Pires-Alves, Melissa¹, Thyagarajan, Baskaran³, Bloom, Jordan E.¹, Gu, Zhengrong¹, Aberle, Karla K.¹, Teymorian, Sasha A.¹, Bannai, Yuka¹, Johnson, Steven C.¹, McArdle, Joseph J.³, Wilson, Brenda A.^1,2
Source:	PEDS: Protein Engineering, Design & Selection. Mar2011, Vol. 24 Issue 3, p247-253. 7p. 3 Color Photographs, 3 Black and White Photographs.
Subjects:	Botulinum toxin, Neurotoxic agents, Antitoxins, Green fluorescent protein, Escherichia coli, Amino acids, Efferent pathways, Laboratory mice
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. [ABSTRACT FROM AUTHOR]
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Database:	Engineering Source

Description
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. [ABSTRACT FROM AUTHOR]
ISSN:	17410126