Two-dimensional crystallization of membrane proteins by reconstitution through dialysis.
Two-dimensional crystallization of membrane proteins by reconstitution through dialysis.
复制标题
DOI：
10.1007/978-1-62703-176-9_3
复制
发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Schmidt-Krey, Ingeborg
中科院分区：
文献类型：
--
作者：
Johnson, Matthew C;Dreaden, Tina M;Schmidt-Krey, Ingeborg
通过二维（2D）结晶和电子晶体学的膜蛋白的研究提供了至关重要的信息的结构和功能的快速增长的这些复杂的蛋白质在一个接近天然的脂质双层。在这里，我们提供了通过透析去除洗涤剂来规划和执行2D结晶试验的方案，包括磷脂的制备和透析设置。讨论了需要考虑的一般因素，如蛋白质制备、增溶洗涤剂、用于重构的脂质和缓冲液条件。几个2D结晶条件突出显示，已显示出很大的希望，在令人惊讶的短时间内生长2D晶体。最后，条件优化的顺序和尺寸的2D晶体的概述。
Studies of membrane proteins by two-dimensional (2D) crystallization and electron crystallography have provided crucial information on the structure and function of a rapidly growing number of these intricate proteins within a close-to-native lipid bilayer. Here we provide protocols for planning and executing 2D crystallization trials by detergent removal through dialysis, including the preparation of phospholipids and the dialysis setup. General factors to be considered, such as the protein preparation, solubilizing detergent, lipid for reconstitution, and buffer conditions are discussed. Several 2D crystallization conditions are highlighted that have shown great promise to grow 2D crystals within a surprisingly short amount of time. Finally, conditions for optimizing order and size of 2D crystals are outlined.