Solubility of the catalytic domains of Botulinum neurotoxin serotype e subtypes

Sheng Chen, Joseph T. Barbieri

Research output: Journal article publicationJournal articleAcademic researchpeer-review

1 Citation (Scopus)

Abstract

The Clostridium botulinum neurotoxins (BoNTs) are the most potent protein toxins known to humans. There are seven serotypes of the BoNTs (A-G), among which serotypes A, B, E and F are known to cause natural human intoxication. To date, eleven subtypes of LC/E, termed E1∼E11, have been identified. The LCs of BoNT/E were insoluble, prohibiting studies towards understanding the mechanisms of toxin action and substrate recognition. In this work, the molecular basis of insolubility of the recombinant LCs of two representative subtypes of BoNT/E, E1(Beluga) and E3 (Alaska), was determined. Hydrophobicity profile and structural modeling predicted a C-terminal candidate region responsible for the insolubility of LC/Es. Deletion of C-terminal 19 residues of LC/E(1-400) resulted in enhanced solubility, from 2 to ∼50% for LC/EAlaskaand from 16 to ∼95% for LC/EBeluga. In addition, resides 230-236 were found to contribute to a different solubility level of LC/EAlaskawhen compared to LC/EBeluga. Substituting residues230TCI232in LC/EAlaskato the corresponding residues of230KYT232in LC/EBelugaenhanced the solubility of LC/EAlaskato a level approaching that of LC/EBeluga. Among these LC/Es and their derivatives, LC/EBeluga1-400 was the most soluble and stable protein. Each LC/E derivative possessed similar catalytic activity, suggesting that the C-terminal region of LC/Es contributed to protein solubility, but not catalytic activity. In conclusion, this study generated a soluble and stable recombinant LC/E and provided insight into the structural components that govern the solubility and stability of the LCs of other BoNT serotypes and Tetanus toxin.
Original languageEnglish
Pages (from-to)18-24
Number of pages7
JournalProtein Expression and Purification
Volume118
DOIs
Publication statusPublished - 1 Feb 2016

Keywords

  • Botulinum neurotoxin E
  • Expression
  • Solubility
  • Subtypes

ASJC Scopus subject areas

  • Biotechnology

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