Mukhina Tetiana, Gerelli Yuri, Hemmerle Arnaud, Koutsioubas Alexandros, Kovalev Kirill, Teulon Jean-Marie, Pellequer Jean-Luc, Daillant Jean, Charitat Thierry, Fragneto Giovanna
Institut Laue-Langevin, 71 av.des Martyrs, BP 156, 38042 Grenoble Cedex, France; Institut Charles Sadron, Université de Strasbourg, CNRS, UPR 22, 67034 Strasbourg, France.
Institut Laue-Langevin, 71 av.des Martyrs, BP 156, 38042 Grenoble Cedex, France; Marche Polytechnic University, Department of Life and Environmental Sciences, Via Brecce Bianche, 60131 Ancona, Italy.
J Colloid Interface Sci. 2021 Sep;597:370-382. doi: 10.1016/j.jcis.2021.03.155. Epub 2021 Mar 31.
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.
通过应用去污剂介导的掺入方法，质子泵跨膜蛋白细菌视紫红质成功地掺入了凝胶相和液相的平面浮动脂质双层中。该方法通过使用石英晶体微天平、原子力显微镜和荧光显微镜技术在单个支撑双层上进行了优化。中子反射和X射线反射技术被用于单个和浮动双层，目的是在亚纳米分辨率下确定蛋白质重构前后该膜蛋白系统的结构和组成。在重构过程中，脂质双层的完整性和蛋白质活性得以保留。观察并表征了由可见光触发的细菌视紫红质功能活性诱导的膜的可逆结构修饰，分辨率达到纳米级。
