Unit

UNIT 5.22 A Bacterial Cell-Free Expression System to Produce Membrane Proteins and Proteoliposomes: From cDNA to Functional Assay

  1. Lavinia Liguori,
  2. Bruno Marques,
  3. Jean-Luc Lenormand

Published Online: 1 NOV 2008

DOI: 10.1002/0471140864.ps0522s54

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Liguori, L., Marques, B. and Lenormand, J.-L. 2008. A Bacterial Cell-Free Expression System to Produce Membrane Proteins and Proteoliposomes: From cDNA to Functional Assay. Current Protocols in Protein Science. 54:5.22:5.22.1–5.22.30.

Author Information

  1. HumProTher Laboratory, TheREx-GREPI, TIMC-IMAG Laboratory, University of Joseph Fourier, UFR de Médecine, La Tronche, France

Publication History

  1. Published Online: 1 NOV 2008
  2. Published Print: NOV 2008

Abstract

Limitations in the production of folded membrane proteins represent the major bottleneck for functional and structural studies of this huge category of macromolecules. Cell-free expression systems provide an attractive alternative to the classical overexpression systems for producing membrane proteins. However, optimization of these systems remains a challenging task, considering the hydrophobic properties of these molecules. This unit describes the production of eukaryotic membrane proteins either in soluble form or integrated into liposomes using a bacterial cell-free expression system. Liposomes in the reaction mixture induce the direct insertion of freshly produced membrane proteins into the bilayer and allow the formation of functional proteoliposomes in which the membrane proteins are correctly folded. Curr. Protoc. Protein Sci. 54:5.22.1-5.22.30. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • cell-free expression system;
  • membrane proteins production;
  • one-step proteoliposomes synthesis;
  • delivery system