A novel complete reconstitution system for membrane integration of the simplest membrane protein

Ken ichi Nishiyama, Masahide Maeda, Masato Abe, Takashi Kanamori, Keiko Shimamoto, Shoichi Kusumoto, Takuya Ueda, Hajime Tokuda

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

A complete reconstitution system for membrane integration of the simplest protein was developed by means of defined factors. A mutant version of Pf3 coat protein, 3L-Pf3 coat, requires neither signal recognition particle/Sec factors nor a membrane potential for its integration into the cytoplasmic membrane of Escherichia coli. Although 3L-Pf3 coat is spontaneously integrated into liposomes composed of phospholipids, diacylglycerol completely blocks such spontaneous integrations at a physiological level. Under the conditions where spontaneous integration does not occur, 3L-Pf3 coat integration was absolutely dependent on a novel integration-stimulating factor. Combination of the PURE system, an in vitro translation system composed of the purified factors involved in translation in E. coli, with liposomes containing the highly purified integration-stimulating factor revealed multiple cycles of 3L-Pf3 coat integration, achieving the complete reconstitution of membrane integration. Based on the function of the factor, we propose that the factor is named MPIase (Membrane Protein Integrase).

Original languageEnglish
Pages (from-to)733-736
Number of pages4
JournalBiochemical and Biophysical Research Communications
Volume394
Issue number3
DOIs
Publication statusPublished - 2010 Apr 9
Externally publishedYes

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Keywords

  • Diacylglycerol
  • Integration-stimulating factor
  • Liposomes
  • Membrane protein integration
  • MPIase
  • PURE system

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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