Production of membrane proteins through the wheat-germ cell-free technology.

Akira Nozawa, Hideaki Nanamiya, Yuzuru Tozawa

Cell-Free Science and Technology Research Center, Ehime University, Ehime, Japan.

Methods in molecular biology (Clifton, N.J.) 2010

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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.

Citation

Akira Nozawa, Hideaki Nanamiya, Yuzuru Tozawa. Production of membrane proteins through the wheat-germ cell-free technology. Methods in molecular biology (Clifton, N.J.). 2010;607:213-8