Efficient production and purification of functional bacteriorhodopsin with a wheat-germ cell-free system and a combination of Fos-choline and CHAPS detergents.
Cell-free translation is one potential approach to the production of functional transmembrane proteins. We have now examined various detergents as supplements to a wheat-germ cell-free system in order to optimize the production and subsequent purification of a functional model transmembrane protein, bacteriorhodopsin. We found that Fos-choline and CHAPS detergents counteracted each other's inhibitory effects on cell-free translation activity and thereby allowed the efficient production and subsequent purification of functional bacteriorhodopsin in high yield.
Genji
Takahisa
T
Cell-Free Science and Technology Research Center and Venture Business Laboratory, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime 790-8577, Japan.
Nozawa
Akira
A
Tozawa
Yuzuru
Y
D001436:Bacteriorhodopsins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000302:isolation & purification*; D002474:Cell-Free System; D002793:Cholic Acids / Q000737:chemistry* / Q000494:pharmacology; D003902:Detergents / Q000737:chemistry* / Q000494:pharmacology; D005854:Germ Cells; D008823:Micelles; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D014176:Protein Biosynthesis; D014908:Triticum
D002793:Cholic Acids; D003902:Detergents; D008823:Micelles; D010767:Phosphorylcholine; D001436:Bacteriorhodopsins; C028810:dodecylphosphocholine; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate