Grafting synthetic transmembrane units to the engineered low-toxicity α-hemolysin to restore its hemolytic activity

Mihoko Ui, Kousuke Harima, Toshiaki Takei, Kouhei Tsumoto, Kazuhito V. Tabata, Hiroyuki Noji, Sumire Endo, Kimio Akiyama, Takahiro Muraoka, Kazushi Kinbara

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

The chemical modification of proteins to provide desirable functions and/or structures broadens their possibilities for use in various applications. Usually, proteins can acquire new functions and characteristics, in addition to their original ones, via the introduction of synthetic functional moieties. Here, we adopted a more radical approach to protein modification, i.e., the replacement of a functional domain of proteins with alternative chemical compounds to build "cyborg proteins." As a proof of concept model, we chose staphylococcal α-hemolysin (Hla), which is a well-studied, pore-forming toxin. The hemolytic activity of Hla mutants was dramatically decreased by truncation of the stem domain, which forms a β-barrel pore in the membrane. However, the impaired hemolytic activity was significantly restored by attaching a pyrenyl-maleimide unit to the cysteine residue that was introduced in the remaining stem domain. In contrast, negatively charged fluorescein-maleimide completely abolished the remaining activity of the mutants.

Original languageEnglish
Pages (from-to)3199-3206
Number of pages8
JournalMolecular BioSystems
Volume10
Issue number12
DOIs
Publication statusPublished - 2014 Dec 1

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology

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