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Crystallization screens: compatibility with the lipidic cubic phase for in meso crystallization of membrane proteins.

机译:结晶筛选:与脂质立方相的相容性,可用于膜蛋白的中观结晶。

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摘要

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 degrees C and at 20 degrees C. 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 degrees C, 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 degrees C 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.
机译:用于生长膜蛋白晶体的中观方法使用自发形成的脂质立方中间相。去污剂增溶的蛋白质与脂质(通常为单油精)分散在一起,从而使立方相自组装。添加沉淀剂以触发晶体成核和生长。商业筛分溶液系列方便用于结晶试验。这项研究的目的是确定Hampton屏幕和Screen 2系列解决方案中的哪一种与in meso方法兼容。这些筛网包含任何可能破坏立方相的成分。 X射线衍射用于相识别和微观结构表征。该研究是在4摄氏度和20摄氏度下进行的。检查了两种样品制备方法。一种使用过量的半强度筛分溶液(制备方法1)。另一个使用了有限数量的未稀释的丝网溶液(制备方法2)。在20摄氏度下,超过90%的筛分溶液会通过制备制备出立方相。 1.使用Prep时,这一数字下降到50%。 2.相反,在制备条件下,在4摄氏度下,有50%到60%的筛子与立方相相容。 1个条件。借助Prep,该数字下降到25%。 2.根据单油精/水系统的相特性,说明了各种筛选组分的作用方式。

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