Fluorinated and hemifluorinated surfactants as alternatives to detergents for membrane protein cell-free synthesis

被引:71
|
作者
Park, Kyu-Ho
Berrier, Catherine
Lebaupain, Florence
Pucci, Bernard
Popot, Jean-Luc
Ghazi, Alexandre
Zito, Francesca [1 ]
机构
[1] CNRS, UMR 7099, Lab Physicochim Mol Membranes Biol, F-75005 Paris, France
[2] Univ Paris 07, CNRS, Inst Biol Physicochim, FRC 550, F-75005 Paris, France
[3] CNRS, UMR 8619, Inst Biochim & Biophys Mol & Cellulaire, F-91405 Orsay, France
[4] Univ Paris 11, F-91405 Orsay, France
[5] Univ Avignon & Pays Vaucluse, Lab Chim Bioorgan & Syst Mol Vectoriels, F-84000 Avignon, France
关键词
detergent; fluorinated surfactant; hemifluorinated surfactant; membrane delivery; membrane protein; MscL;
D O I
10.1042/BJ20061473
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Hemifluorinated and fluorinated surfactants are lipophobic and, as such, non-detergent. Although they do not solubilize biological membranes, they can, after conventional solubilization, substitute for detergents to keep membrane proteins soluble, which generally improves their stability [Breyton, Chabaud, Chaudier, Pucci and Popot (2004) FEBS Lett. 564, 312-318]. In the present study, we show that (hemi)fluorinated surfactants can be used for in vitro synthesis of membrane proteins: they do not interfere with protein synthesis, and they provide a suitable environment for MscL, a pentameric mechanosensitive channel, to fold and oligomerize to its native functional state. Following synthesis, both types of surfactants can be used to deliver MscL directly to pre-formed lipid vesicles. The electrophysiological activity of MscL synthesized in vitro in the presence of either hemi- or per-fluorinated surfactant is similar to that of the protein expressed in vivo.
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页码:183 / 187
页数:5
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