Abstract
Membrane proteins play crucial roles in various processes. However, biochemical characterization of the membrane proteins remains challenging due to the difficulty in producing membrane proteins in a functional state. Here, we describe a novel method for the production of functional membrane proteins based on a wheat germ cell-free translation system. Using this method, functional membrane proteins are successfully synthesized in the presence of liposomes and a detergent. In addition, the synthesized membrane proteins are easily purified from the cell-free translation mixture as proteoliposomes by sucrose density gradient ultracentrifugation. These advantages over conventional approaches are very helpful for the clarification of the function of membrane proteins.
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Acknowledgments
This study was supported by a Grant-in-Aid for Scientific Research on Priority Areas (Plant Membrane Transport) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (no. 18056016 to Y.T.)
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Nozawa, A., Nanamiya, H., Tozawa, Y. (2010). Production of Membrane Proteins Through the Wheat-Germ Cell-Free Technology. In: Endo, Y., Takai, K., Ueda, T. (eds) Cell-Free Protein Production. Methods in Molecular Biology, vol 607. Humana Press. https://doi.org/10.1007/978-1-60327-331-2_17
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DOI: https://doi.org/10.1007/978-1-60327-331-2_17
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