Conformational Chaperones for Structural Studies of Membrane Proteins Using Antibody Phage Display with Nanodiscs.
Conformational Chaperones for Structural Studies of Membrane Proteins Using Antibody Phage Display with Nanodiscs.
复制标题
使用纳米散发的抗体噬菌体显示膜蛋白的结构研究构象伴侣。
DOI：
10.1016/j.str.2015.11.014
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发表时间：
2016-02-02
期刊：
影响因子：
--
通讯作者：
Kossiakoff AA
中科院分区：
文献类型：
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作者：
Dominik PK;Borowska MT;Dalmas O;Kim SS;Perozo E;Keenan RJ;Kossiakoff AA
膜生物物理学中的一个主要挑战是定义蛋白质的构象转变与其功能之间的机械联系。我们描述了一种新的方法来稳定膜蛋白在天然脂质环境中的瞬时功能状态，允许它们的结构和生化特征。这是通过将基于抗体Fab的噬菌体展示选择的能力与将膜蛋白靶包埋在脂质填充的纳米盘中的益处相结合来实现的。除了提供稳定的脂质环境之外，纳米盘提供了优于基于去污剂的形式的显著技术优势。这使得能够产生丰富的高性能Fab结合剂库，其可用作结晶伴侣，用作单颗粒cryo-EM的基准标记物和用作不同构象状态的探针。此外，纳米盘产生的Fab可用于鉴定最佳模拟天然膜环境的去污剂以用于生物物理研究。
A major challenge in membrane biophysics is to define the mechanistic linkages between a protein’s conformational transitions and its function. We describe a novel approach to stabilize transient functional states of membrane proteins in native-like lipid environments allowing for their structural and biochemical characterization. This is accomplished by combining the power of antibody Fab-based phage display selection with the benefits of embedding membrane protein targets in lipid-filled nanodiscs. In addition to providing a stabilizing lipid environment, nanodiscs afford significant technical advantages over detergent-based formats. This enables the production of a rich pool of high performance Fab binders that can be used as crystallization chaperones, as fiducial markers for single particle cryo-EM and as probes of different conformational states. Moreover, nanodisc generated Fabs can be used to identify detergents that best mimic native membrane environments for use in biophysical studies.