Crystallization screens:: Compatibility with the lipidic cubic phase for in meso crystallization of membrane proteins
Crystallization screens:: Compatibility with the lipidic cubic phase for in meso crystallization of membrane proteins
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DOI：
10.1016/s0006-3495(01)75694-9
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发表时间：
2001-07-01
影响因子：
3.4
通讯作者：
Caffrey, M
中科院分区：
文献类型：
--
作者：
Cherezov, V;Fersi, H;Caffrey, M
生长膜蛋白晶体的内介孔方法使用自发形成的脂质立方介孔相。洗涤剂溶解的蛋白质与脂质分散，通常是单油质，这样立方相就会自组装。加入沉淀剂以触发晶体成核和生长。商用筛液系列便于在结晶试验中使用。本研究的目的是确定哪些汉普顿筛选和筛选2系列的解决方案是兼容的内介观方法。这些筛网含有的任何一种成分都可能破坏立方相。x射线衍射用于物相鉴定和微观结构表征。研究是在4度和20度的温度下进行的。研究了两种类型的样品制备。其中一个使用了过量的半强度筛选溶液（Prep. 1）。另一组使用限量的未稀释筛选液（Prep. 2）。在20°c时，超过90%的筛分溶液产生了Prep. 1的立方相。使用Prep. 2后，这一数字降至50%。相比之下，在Prep. 1条件下，50%至60%的筛片在4度下与立方相兼容。使用Prep. 2后，这一数字降至25%。根据单油/水体系的相性质，解释了不同筛分组分的作用方式。
The in meso method for growing crystals of membrane proteins uses a spontaneously forming lipidic cubic mesophase. The detergent-solubilized protein is dispersed with lipid, typically monoolein, and in so doing the cubic phase self-assembles. A precipitant is added to trigger crystal nucleation and growth. The commercial screen solution series are convenient for use in crystallization trials. The aim of this study was to determine which of the Hampton Screen and Screen 2 series of solutions are compatible with the in meso method. These screens contain components any of which could destroy the cubic phase. X-ray diffraction was used for phase identification and for microstructure characterization. The study was done at 4 degreesC and at 20 degreesC. Two types of sample preparations were examined. One used an excess of half-strength screen solution (Prep. 1). The other used a limiting quantity of undiluted screen solution (Prep. 2). At 20 degreesC, over 90% of the screen solutions produced the cubic phase with Prep. 1. This figure dropped to 50% with Prep. 2. In contrast, 50 to 60% of the screens were cubic phase compatible at 4 degreesC under Prep. 1 conditions. The figure fell to 25% with Prep. 2. The mode of action of the diverse screen components are explained on the basis of the phase properties of the monoolein/water system.