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Insertion and activation of functional Bacteriorhodopsin in a floating bilayer

Tetiana Mukhina, Yuri Gerelli, Arnaud Hemmerle, Alexandros Koutsioubas, Kirill Kovalev, Jean-MarieTeulon, Jean-Luc Pellequer, Jean Daillant, Thierry Charitat, Giovanna Fragneto
Journal of Colloid and Interface Science 2021, Volume 597, September 2021, Pages 370-382

The proton pump transmembrane protein bacteriorhodopsin was successfully incorporated into planar floating lipid bilayers in gel and fluid phases, by applying a detergent-mediated incorporation method. The method was optimized on single supported bilayers by using quartz crystal microbalance, atomic force and fluorescence microscopy techniques. Neutron and X-ray reflectometry were used on both single and floating bilayers with the aim of determining the structure and composition of this membrane-protein system before and after protein reconstitution at sub-nanometer resolution. Lipid bilayer integrity and protein activity were preserved upon the reconstitution process. Reversible structural modifications of the membrane, induced by the bacteriorhodopsin functional activity triggered by visible light, were observed and characterized at the nanoscale.

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