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用于功能分析的含光合天线蛋白的束缚脂质双层的构建和结构分析。

Construction and structural analysis of tethered lipid bilayer containing photosynthetic antenna proteins for functional analysis.

机构信息

Department of Frontier Materials, Graduate School of Engineering, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan.

出版信息

Biomacromolecules. 2011 Jul 11;12(7):2850-8. doi: 10.1021/bm200585y. Epub 2011 Jun 22.

Abstract

The construction and structural analysis of a tethered planar lipid bilayer containing bacterial photosynthetic membrane proteins, light-harvesting complex 2 (LH2), and light-harvesting core complex (LH1-RC) is described and establishes this system as an experimental platform for their functional analysis. The planar lipid bilayer containing LH2 and/or LH1-RC complexes was successfully formed on an avidin-immobilized coverglass via an avidin-biotin linkage. Atomic force microscopy (AFM) showed that a smooth continuous membrane was formed there. Lateral diffusion of these membrane proteins, observed by a fluorescence recovery after photobleaching (FRAP), is discussed in terms of the membrane architecture. Energy transfer from LH2 to LH1-RC within the tethered membrane was observed by steady-state fluorescence spectroscopy, indicating that the tethered membrane can mimic the natural situation.

摘要

描述了一种含细菌光合膜蛋白、光捕获复合物 2(LH2)和光捕获核心复合物(LH1-RC)的系绳平面脂质双层的构建和结构分析,并将该系统确立为其功能分析的实验平台。通过亲和素-生物素连接,成功地在固定有亲和素的盖玻片上形成含有 LH2 和/或 LH1-RC 复合物的平面脂质双层。原子力显微镜(AFM)显示,在那里形成了一个光滑连续的膜。通过光漂白后荧光恢复(FRAP)观察到这些膜蛋白的侧向扩散,并根据膜结构进行了讨论。通过稳态荧光光谱观察到在系绳膜内从 LH2 到 LH1-RC 的能量转移,表明系绳膜可以模拟自然情况。

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