Efficient production and purification of functional bacteriorhodopsin with a wheat-germ cell-free system and a combination of Fos-choline and CHAPS detergents

Biochem Biophys Res Commun. 2010 Oct 1;400(4):638-42. doi: 10.1016/j.bbrc.2010.08.119. Epub 2010 Aug 31.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriorhodopsins / antagonists & inhibitors
  • Bacteriorhodopsins / biosynthesis*
  • Bacteriorhodopsins / isolation & purification*
  • Cell-Free System
  • Cholic Acids / chemistry*
  • Cholic Acids / pharmacology
  • Detergents / chemistry*
  • Detergents / pharmacology
  • Germ Cells
  • Micelles
  • Phosphorylcholine / analogs & derivatives*
  • Phosphorylcholine / chemistry
  • Phosphorylcholine / pharmacology
  • Protein Biosynthesis*
  • Triticum

Substances

  • Cholic Acids
  • Detergents
  • Micelles
  • Phosphorylcholine
  • Bacteriorhodopsins
  • dodecylphosphocholine
  • 3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate