Direct preparation of giant proteo-liposomes by in vitro membrane protein synthesis

Shin ichiro M. Nomura, Satoshi Kondoh, Wakiko Asayama, Akikazu Asada, Shigemichi Nishikawa, Kazunari Akiyoshi

Research output: Contribution to journalArticlepeer-review

62 Citations (Scopus)

Abstract

We investigated the direct constitution of membrane proteins into giant liposomes in cell-free (in vitro) protein synthesis. Giant liposomes were present in a translation reaction cocktail of a wheat germ cell-free protein translation system. Apo cytochrome b5 (b5) and its fusion proteins were synthesized and directly localized in the liposomes. After the translation reaction, the proteo-liposomes were isolated by simplified discontinuous density-gradient centrifugation. Apo cytochrome b5 conjugated dihydrofolate reductase (DHFR) was synthesized in the same procedure and the protein was directly displayed on the liposome surface. b5 acts as a "hydrophobic tag" for recruitment to the liposome surface.

Original languageEnglish
Pages (from-to)190-195
Number of pages6
JournalJournal of Biotechnology
Volume133
Issue number2
DOIs
Publication statusPublished - 2008 Jan 20
Externally publishedYes

Keywords

  • Cytochrome b5
  • Giant liposome
  • In vitro protein synthesis
  • Membrane protein
  • Wheat germ lysate

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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