Liposome chaperon in cell-free membrane protein synthesis: One-step preparation of KcsA-integrated liposomes and electrophysiological analysis by the planar bilayer method

M. Ando, M. Akiyama, D. Okuno, M. Hirano, Toru Ide, S. Sawada, Y. Sasaki, K. Akiyoshi

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Abstract

Chaperoning functions of liposomes were investigated using cell-free membrane protein synthesis. KcsA potassium channel-reconstituted liposomes were prepared directly using cell-free protein synthesis. In the absence of liposomes, all synthesized KcsA protein aggregated. In the presence of liposomes, however, synthesized KcsA spontaneously integrated into the liposome membrane. The KscA-reconstituted liposomes were transferred to the planar bilayer across a small hole in a thin plastic sheet and the channel function of KcsA was examined. The original electrophysiological activities, such as voltage- and pH-dependence, were observed. These results suggested that in cell-free membrane protein synthesis, liposomes act as chaperones, preventing aggregation and assisting in folding and tetrameric formation, thereby allowing full channel activity.

Original languageEnglish
Pages (from-to)258-264
Number of pages7
JournalBiomaterials Science
Volume4
Issue number2
DOIs
Publication statusPublished - Feb 1 2016

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ASJC Scopus subject areas

  • Biomedical Engineering
  • Materials Science(all)

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