Chapter 11 - Reconstitution of membrane proteins in phospholipid bilayer nanodiscs.
Chapter 11 - Reconstitution of membrane proteins in phospholipid bilayer nanodiscs.
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DOI：
10.1016/s0076-6879(09)64011-8
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发表时间：
2009
影响因子：
--
通讯作者：
Sligar, S. G.
中科院分区：
文献类型：
--
作者：
Ritchie, T. K.;Grinkova, Y. V.;Bayburt, T. H.;Denisov, I. G.;Zolnerciks, J. K.;Atkins, W. M.;Sligar, S. G.
自组装磷脂双层纳米盘已成为模型膜系统中功能重组膜蛋白的重要工具。纳米圆盘由载脂蛋白A-1支架蛋白的工程衍生物包裹的脂质结构域组成，可以定制成具有不同直径的圆盘的均匀制备，并具有用于各种纯化策略的表位标签。靶膜自组装成纳米圆盘的一个关键方面在于脂质：蛋白质比例的优化。在这里，我们描述了进行这种优化的策略，并提供了将细菌视紫红质重组为三聚体、视紫红质和功能活性p糖蛋白的例子。总之，这些都证明了纳米盘技术在制备结构广泛的膜蛋白单分散样品方面的多功能性。
Self-assembled phospholipid bilayer Nanodiscs have become an important and versatile tool among model membrane systems to functionally reconstitute membrane proteins. Nanodiscs consist of lipid domains encased within an engineered derivative of apolipoprotein A-1 scaffold proteins, which can be tailored to yield homogeneous preparations of disks with different diameters, and with epitope tags for exploitation in various purification strategies. A critical aspect of the self-assembly of target membranes into Nanodiscs lies in the optimization of the lipid:protein ratio. Here we describe strategies for performing this optimization and provide examples for reconstituting bacteriorhodpsin as a trimer, rhodopsin, and functionally active P-glycoprotein. Together these demonstrate the versatility of Nanodisc technology for preparing monodisperse samples of membrane proteins of wide-ranging structure.