Fluorinated and hemifluorinated surfactants as alternatives to detergents for membrane protein cell-free synthesis
Résumé
Hemifluorinated and fluorinated surfactants are lipophobic and, as such, non-detergent. While they do not solubilise biological membranes, they can, after conventional solubilisation, substitute to detergents to keep membrane proteins soluble, which generally improves their stability (Breyton et al. FEBS Lett. 564, 312-318, 2004). Here, 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 directly MscL to preformed lipid vesicles. The electrophysiological activity of MscL synthesized in vitro in the presence of either hemi- or perfluorinated surfactant is similar to that of the protein expressed in vivo.
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