期刊
JOURNAL OF BIOENERGETICS AND BIOMEMBRANES
卷 42, 期 3, 页码 235-240出版社
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s10863-010-9285-8
关键词
E. coli cell-free system; Heme A; Paracoccus denitrificans
资金
- MEXT [17GS0429, 16087026]
- Keio University
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.
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