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.
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Katayama, Y., Shimokata, K., Suematsu, M. et al. Cell-free synthesis of cytochrome c oxidase, a multicomponent membrane protein. J Bioenerg Biomembr 42, 235–240 (2010). https://doi.org/10.1007/s10863-010-9285-8
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DOI: https://doi.org/10.1007/s10863-010-9285-8