Premium
Hemifluorinated surfactants: a non‐dissociating environment for handling membrane proteins in aqueous solutions?
Author(s) -
Breyton Cécile,
Chabaud Elodie,
Chaudier Yann,
Pucci Bernard,
Popot Jean-Luc
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(04)00227-3
Subject(s) - chemistry , bacteriorhodopsin , membrane protein , cytochrome c , micelle , chromatography , aqueous solution , membrane , moiety , cytochrome , tris , octyl glucoside , integral membrane protein , transmembrane protein , biochemistry , organic chemistry , enzyme , mitochondrion , receptor
The instability of membrane proteins in detergent solution can generally be traced to the dissociating character of detergents and often correlates with delipidation. We examine here the possibility of substituting detergents, after membrane proteins have been solubilized, with non‐detergent surfactants whose hydrophobic moiety contains a perfluorinated region that makes it lipophobic. In order to improve its affinity for the protein surface, the fluorinated chain is terminated by an ethyl group. Test proteins included bacteriorhodopsin, the cytochrome b 6 f complex, and the transmembrane region of the bacterial outer membrane protein OmpA. All three proteins were purified using classical detergents and transferred into solutions of C 2 H 5 C 6 F 12 C 2 H 4 ‐ S ‐poly‐ Tris ‐(hydroxymethyl)aminomethane (HF‐TAC). Transfer to HF‐TAC maintained the native state of the proteins and prevented their precipitation. Provided the concentration of HF‐TAC was high enough, HF‐TAC/membrane protein complexes ran as single bands upon centrifugation in sucrose gradients. Bacteriorhodopsin and the cytochrome b 6 f complex, both of which are detergent‐sensitive, exhibited increased biochemical stability upon extended storage in the presence of a high concentration of HF‐TAC as compared to detergent micelles. The stabilization of cytochrome b 6 f is at least partly due to a better retention of protein‐bound lipids.