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

Research output: Contribution to journalArticle

8 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)235-240
Number of pages6
JournalJournal of Bioenergetics and Biomembranes
Volume42
Issue number3
DOIs
Publication statusPublished - 2010 Jun

Fingerprint

Electron Transport Complex IV
Membrane Proteins
Proteins
Paracoccus denitrificans
Amino Acids
Cell-Free System
Protein Subunits
Cytochromes c
Heme
Isotopes
Oxidation-Reduction
Copper

Keywords

  • E. coli cell-free system
  • Heme A
  • Paracoccus denitrificans

ASJC Scopus subject areas

  • Physiology
  • Cell Biology
  • Medicine(all)

Cite this

Katayama, Y., Shimokata, K., Suematsu, M., Ogura, T., Tsukihara, T., Yoshikawa, S., & Shimada, H. (2010). Cell-free synthesis of cytochrome c oxidase, a multicomponent membrane protein. Journal of Bioenergetics and Biomembranes, 42(3), 235-240. https://doi.org/10.1007/s10863-010-9285-8

Cell-free synthesis of cytochrome c oxidase, a multicomponent membrane protein. / Katayama, Yukie; Shimokata, Kunitoshi; Suematsu, Makoto; Ogura, Takashi; Tsukihara, Tomitake; Yoshikawa, Shinya; Shimada, Hideo.

In: Journal of Bioenergetics and Biomembranes, Vol. 42, No. 3, 06.2010, p. 235-240.

Research output: Contribution to journalArticle

Katayama, Y, Shimokata, K, Suematsu, M, Ogura, T, Tsukihara, T, Yoshikawa, S & Shimada, H 2010, 'Cell-free synthesis of cytochrome c oxidase, a multicomponent membrane protein', Journal of Bioenergetics and Biomembranes, vol. 42, no. 3, pp. 235-240. https://doi.org/10.1007/s10863-010-9285-8
Katayama, Yukie ; Shimokata, Kunitoshi ; Suematsu, Makoto ; Ogura, Takashi ; Tsukihara, Tomitake ; Yoshikawa, Shinya ; Shimada, Hideo. / Cell-free synthesis of cytochrome c oxidase, a multicomponent membrane protein. In: Journal of Bioenergetics and Biomembranes. 2010 ; Vol. 42, No. 3. pp. 235-240.
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