HiLiDe鈥擲ystematic Approach to Membrane Protein Crystallization in Lipid and Detergent

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• 作者：Pontus Gourdon ; Jacob Lauwring Andersen ; Kim Langmach Hein ; Maike Bublitz ; Bj酶rn Panyella Pedersen ; Xiang-Yu Liu ; Laure Yatime ; Maria Nyblom ; Thorbj酶rn Terndrup Nielsen ; Claus Olesen ; Jesper Vuust M酶ller ; Poul Nissen ; Jens Preben Morth
• 刊名：Crystal Growth & Design
• 出版年：2011
• 出版时间：June 1, 2011
• 年：2011
• 卷：11
• 期：6
• 页码：2098-2106
• 全文大小：1116K
• 年卷期：v.11,no.6(June 1, 2011)
• ISSN：1528-7505

文摘

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.