Journal of Bioenergetics and Biomembranes

, Volume 42, Issue 3, pp 235–240

Cell-free synthesis of cytochrome c oxidase, a multicomponent membrane protein

  • Yukie Katayama
  • Kunitoshi Shimokata
  • Makoto Suematsu
  • Takashi Ogura
  • Tomitake Tsukihara
  • Shinya Yoshikawa
  • Hideo Shimada
Article

Abstract

Cell-free protein synthesis is a useful technique that can site-specifically incorporate isotope-labeled amino acids into proteins. This incorporation is essential for infrared analyses of the electronic state of a specific amino acid residue used to elucidate protein function. Although 17 membrane proteins have been synthesized in their active state by cell-free systems, to date no hetero-subunit protein has been synthesized with this technique, suggesting that there are serious technical limitations. Here we report the cell-free synthesis of Paracoccus denitrificans cytochrome c oxidase, a membrane protein complex composed of three distinct subunits that contain two heme A molecules and two redox-active copper centers. The synthesized protein exhibited normal Soret/vis absorption spectra and ferrocytochrome c oxidation activity.

Keywords

E. coli cell-free system Heme A Paracoccus denitrificans 

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Yukie Katayama
    • 1
  • Kunitoshi Shimokata
    • 2
  • Makoto Suematsu
    • 3
  • Takashi Ogura
    • 1
    • 4
  • Tomitake Tsukihara
    • 1
  • Shinya Yoshikawa
    • 1
    • 4
  • Hideo Shimada
    • 1
  1. 1.Picobiology Institute, Graduate School of Life Science, Center of Advanced Science and TechnologyUniversity of HyogoAkohJapan
  2. 2.World Intec Co. LTDChyouJapan
  3. 3.Department of Biochemistry and Integrative Medical Biology, School of MedicineKeio UniversityShinjukuJapan
  4. 4.Graduate School of Life ScienceUniversity of HyogoAkohJapan

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