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

Détails

ID Serval
serval:BIB_71C8C6A79965
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Wheat germ cell-free expression: Two detergents with a low critical micelle concentration allow for production of soluble HCV membrane proteins.
Périodique
Protein Expression and Purification
Auteur(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
Statut éditorial
Publié
Date de publication
2015
Peer-reviewed
Oui
Volume
105
Pages
39-46
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Résumé
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.
Mots-clé
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
Création de la notice
02/01/2015 10:26
Dernière modification de la notice
20/08/2019 15:30
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