Production of membrane proteins through the wheat-germ cell-free technology.
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.
Nozawa
Akira
A
Cell-Free Science and Technology Research Center, Ehime University, Ehime, Japan.
Nanamiya
Hideaki
H
Tozawa
Yuzuru
Y
D029681:Arabidopsis Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D003902:Detergents / Q000737:chemistry; D008081:Liposomes; D026901:Membrane Transport Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014176:Protein Biosynthesis; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012639:Seeds / Q000378:metabolism; D014908:Triticum / Q000196:embryology / Q000235:genetics / Q000378:metabolism*
D029681:Arabidopsis Proteins; D003902:Detergents; D008081:Liposomes; D026901:Membrane Transport Proteins; D011994:Recombinant Proteins; C481826:phosphate translocator 1, Arabidopsis