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Biochem J. 2007 Apr 1;403(1):183-7.

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

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1
Laboratoire de Physico-chimie Moléculaire des Membranes Biologiques, UMR 7099, CNRS and Université Paris-7, Institut de Biologie Physico-Chimique, CNRS FRC 550, F-75005 Paris, France.

Abstract

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.

PMID:
17176254
PMCID:
PMC1828882
DOI:
10.1042/BJ20061473
[Indexed for MEDLINE]
Free PMC Article
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