A bacterial cell-free expression system to produce membrane proteins and proteoliposomes: from cDNA to functional assay.
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.
Liguori
Lavinia
L
HumProTher Laboratory, TheREx-GREPI, TIMC-IMAG Laboratory, University of Joseph Fourier, UFR de Médecine, La Tronche, France.
Marques
Bruno
B
HumProTher Laboratory, TheREx-GREPI, TIMC-IMAG Laboratory, University of Joseph Fourier, UFR de Médecine, La Tronche, France.
Lenormand
Jean-Luc
JL
HumProTher Laboratory, TheREx-GREPI, TIMC-IMAG Laboratory, University of Joseph Fourier, UFR de Médecine, La Tronche, France.
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism*; D004586:Electrophoresis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism*; D017510:Protein Folding; D011510:Proteolipids / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis
D018076:DNA, Complementary; D008565:Membrane Proteins; D011510:Proteolipids; D011994:Recombinant Proteins; C020485:proteoliposomes