論文

査読有り 国際誌
2023年1月24日

Molecular engineering of a minimal E-cadherin inhibitor protein derived from Clostridium botulinum hemagglutinin.

The Journal of biological chemistry
  • Sho Amatsu
  • ,
  • Takuhiro Matsumura
  • ,
  • Masahiko Zuka
  • ,
  • Yukako Fujinaga

299
3
開始ページ
102944
終了ページ
102944
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.jbc.2023.102944

Hemagglutinin (HA), a non-toxic component of the botulinum neurotoxin (BoNT) complex, binds to E-cadherin and inhibits E-cadherin-mediated cell-cell adhesion. HA is a 470 kDa protein complex comprising six HA1, three HA2, and three HA3 subcomponents. Thus, to prepare recombinant full-length HA in vitro, it is necessary to reconstitute the macromolecular complex from purified HA subcomponents, which involves multiple purification steps. In this study, we developed NanoHA, a minimal E-cadherin inhibitor protein derived from Clostridium botulinum HA with a simple purification strategy needed for production. NanoHA, containing HA2 and a truncated mutant of HA3 (aa 380-626; termed as HA3mini), is a 47 kDa single polypeptide (one-tenth the molecular weight of full-length HA, 470 kDa) engineered with three types of modifications: (i) a short linker sequence between the C-terminus of HA2 and N-terminus of HA3, (ii) a chimeric complex composed of HA2 derived from the serotype C BoNT complex and HA3mini from the serotype B BoNT complex; and (iii) three amino acid substitutions from hydrophobic to hydrophilic residues on the protein surface. We demonstrated that NanoHA inhibits E-cadherin-mediated cell-cell adhesion of epithelial cells (e.g., Caco-2 and MDCK cells) and disrupts their epithelial barrier. Finally, unlike full-length HA, NanoHA can be transported from the basolateral side to adherens junctions via passive diffusion. Overall, these results indicate that the rational design of NanoHA provides a minimal E-cadherin inhibitor with a wide variety of applications as a lead molecule and for further molecular engineering.

リンク情報
DOI
https://doi.org/10.1016/j.jbc.2023.102944
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/36707052
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9958082
ID情報
  • DOI : 10.1016/j.jbc.2023.102944
  • PubMed ID : 36707052
  • PubMed Central 記事ID : PMC9958082

エクスポート
BibTeX RIS