Reconstitution and imaging of a membrane protein in a nanometer-size phospholipid bilayer
Reconstitution and imaging of a membrane protein in a nanometer-size phospholipid bilayer
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DOI：
10.1006/jsbi.1998.4007
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发表时间：
1998-09-01
影响因子：
3
通讯作者：
Sligar, SG
中科院分区：
文献类型：
--
作者：
Bayburt, TH;Carlson, JW;Sligar, SG
利用纳米尺度的磷脂双分子层作为载体，研究了功能单膜蛋白的重构。该纳米层由一个直径约10纳米的圆形磷脂结构域组成，该结构域由载脂蛋白A1稳定。作为该方法的演示，我们在肝微粒体nadph -细胞色素P450还原酶存在的情况下形成了纳米层。将含有酶的纳米层溶液与新切割的云母基质孵育，可自发形成完全定向的盘状磷脂结构域支持单层。定向单层中的p450 -还原酶保持其催化活性。通过扫描力显微镜的表征显示，分离的单膜蛋白可以随着时间的推移稳定成像。这些结果定义了一种在双层环境中研究单膜蛋白的新技术。(C) 1998学术出版社。
A phospholipid bilayer of nanometer dimension has been used as a support for the study of reconstituted functional single-membrane proteins. This nanobilayer consists of an approximately 10-nm-diameter circular phospholipid domain stabilized by apolipoprotein A1. As a demonstration of this methodology, we formed the nanobilayers in the presence of hepatic microsomal NADPH-cytochrome P450 reductase. Incubation of a solution of enzyme-containing nanobilayers with a freshly cleaved mica substrate resulted in the spontaneous formation of a fully oriented supported monolayer of discoidal phospholipid domains. The P450-reductase in the oriented monolayer retains its catalytic activity. Characterization by scanning force microscopy revealed isolated single-membrane proteins that could be stably imaged over time. These results define a novel technique for the study of single-membrane proteins in a bilayer environment. (C) 1998 Academic Press.