HiLiDe-Systematic Approach to Membrane Protein Crystallization in Lipid and Detergent
HiLiDe-Systematic Approach to Membrane Protein Crystallization in Lipid and Detergent
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DOI：
10.1021/cg101360d
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发表时间：
2011-06-01
影响因子：
3.8
通讯作者：
Morth, Jens Preben
中科院分区：
文献类型：
--
作者：
Gourdon, Pontus;Andersen, Jacob Lauwring;Morth, Jens Preben
获得初始结晶命中和后续优化仍然是膜蛋白结构测定的主要瓶颈。在这里，我们描述了一种在高浓度脂质和洗涤剂（HILIDE）中实现衍射质量膜蛋白晶体的方法。通过控制膜蛋白/脂质/洗涤剂比例的变化，实现初始命中，并通过采用适合该方法的浓度的二级洗涤剂进行系统筛选，建立优化的晶体。该方法一般适用于天然和异种表达的膜蛋白，并与任何结晶方法兼容，如油下蒸汽扩散和批量结晶。该方法提供了用户定义的脂质环境，并且本身不限于具有特定性质的脂质，例如脂相结晶方法。
To obtain the initial crystallization hit and the subsequent optimization remain the major bottlenecks in membrane protein structure determination. Here we describe an approach to achieve diffraction quality membrane protein crystals in high concentrations of lipid and detergent (HILIDE). Initial hits are achieved :through a controlled variation of the membrane protein/lipid/detergent ratios, and optimized crystals are established by systematic screening with secondary detergents at concentrations that are adapted for the method. The approach is generally applicable for native and heterologously expressed membrane proteins, and it is compatible with any crystallization method, such as vapor diffusion and batch crystallization under oil. The method provides a user defined lipidic environment and is in itself not limited to lipids with particular properties, such as the lipid-phase crystallization methods.