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

Methods Mol Biol. 2010;607:213-8. doi: 10.1007/978-1-60327-331-2_17.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins / biosynthesis*
  • Arabidopsis Proteins / chemistry
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / isolation & purification
  • Cell-Free System
  • Centrifugation, Density Gradient
  • Detergents / chemistry
  • Liposomes
  • Membrane Transport Proteins / biosynthesis*
  • Membrane Transport Proteins / chemistry
  • Membrane Transport Proteins / genetics
  • Membrane Transport Proteins / isolation & purification
  • Protein Biosynthesis
  • Protein Engineering / methods*
  • Recombinant Proteins / biosynthesis
  • Seeds / metabolism
  • Triticum / embryology
  • Triticum / genetics
  • Triticum / metabolism*

Substances

  • Arabidopsis Proteins
  • Detergents
  • Liposomes
  • Membrane Transport Proteins
  • Recombinant Proteins
  • phosphate translocator 1, Arabidopsis