Wheat germ cell-free expression: Two detergents with a low critical micelle concentration allow for production of soluble HCV membrane proteins.

Details

Serval ID
serval:BIB_71C8C6A79965
Type
Article: article from journal or magazin.
Collection
Publications
Title
Wheat germ cell-free expression: Two detergents with a low critical micelle concentration allow for production of soluble HCV membrane proteins.
Journal
Protein Expression and Purification
Author(s)
Fogeron M.L., Badillo A., Jirasko V., Gouttenoire J., Paul D., Lancien L., Moradpour D., Bartenschlager R., Meier B.H., Penin F., Böckmann A.
ISSN
1096-0279 (Electronic)
ISSN-L
1046-5928
Publication state
Published
Issued date
2015
Peer-reviewed
Oui
Volume
105
Pages
39-46
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
Membrane proteins are notoriously difficult to express in a soluble form. Here, we use wheat germ cell-free expression in the presence of various detergents to produce the non-structural membrane proteins 2, 4B and 5A of the hepatitis C virus (HCV). We show that lauryl maltose neopentyl glycol (MNG-3) and dodecyl octaethylene glycol ether (C12E8) detergents can yield essentially soluble membrane proteins at detergent concentrations that do not inhibit the cell-free reaction. This finding can be explained by the low critical micelle concentration (CMC) of these detergents, which keeps the monomer concentrations low while at the same time providing the necessary excess of detergent concentration above CMC required for full target protein solubilization. We estimate that a tenfold excess of detergent micelles with respect to the protein concentration is sufficient for solubilization, a number that we propose as a guideline for detergent screening assays.
Keywords
Cell-Free System, Detergents/chemistry, Hepacivirus/genetics, Membrane Proteins/genetics, Membrane Proteins/metabolism, Micelles, Recombinant Proteins/genetics, Recombinant Proteins/metabolism, Triticum/metabolism, Viral Proteins/genetics, Viral Proteins/metabolism
Pubmed
Web of science
Create date
02/01/2015 10:26
Last modification date
20/08/2019 15:30
Usage data