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

Mengfei Ho, Li Hsin Chang, Melissa Pires-Alves, Baskaran Thyagarajan, Jordan E. Bloom, Zhengrong Gu, Karla K. Aberle, Sasha A. Teymorian, Yuka Bannai, Steven C. Johnson, Joseph J. McArdle, Brenda A. Wilson

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)247-253
Number of pages7
JournalProtein Engineering, Design and Selection
Volume24
Issue number3
DOIs
StatePublished - Mar 2011

Keywords

  • antitoxin
  • cargo
  • delivery vehicle
  • neurotoxin

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Molecular Biology

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