Production of multi-subunit complexes on liposome through an E. coli cell-free expression system.

Yutetsu Kuruma, Toshiharu Suzuki, Takuya Ueda

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Recently cell-free translation systems became a laboratory research tool to obtain an objective protein. However, a general method for in vitro protein synthesis and multi-subunit complex formation on lipid membrane has not been established. Here, we describe the procedure for the production of subcomplexes of F(o)F(1)-ATP synthase using a reconstructed cell-free translation system. As for the membrane part F(o) (a(1)b(2)c((10-15))), cosynthesis of c-subunit and UncI proteins, which are integral membrane proteins, and further c (11)-ring formation in lipid bilayer are performed by use of a liposomes-containing cell-free system. Moreover, as for the cytoplasm part F(1) (alpha(3)beta(3)gammadeltaepsilon), subcomplex formations are successfully achieved by mixing the translation products.

Original languageEnglish
Pages (from-to)161-171
Number of pages11
JournalMethods in molecular biology (Clifton, N.J.)
Volume607
DOIs
Publication statusPublished - 2010

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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