Crystallizing membrane proteins using lipidic mesophases.
Crystallizing membrane proteins using lipidic mesophases.
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DOI：
10.1038/nprot.2009.31
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发表时间：
2009
期刊：
影响因子：
14.8
通讯作者：
中科院分区：
文献类型：
--
作者：
详细的协议，使结晶膜蛋白，利用介晶中间相描述。这已经被不同地称为脂质立方相或内消旋方法。该方法已被证明是相当普遍的，因为它已被用于解决原核和真核蛋白质的X射线晶体结构，蛋白质是单体，同源和异源多聚体，含发色团和无发色团，和α-螺旋和β-桶蛋白质。它最近的成功是人类工程β2-肾上腺素能和腺苷A2 A G蛋白偶联受体。提供了用于制备和表征介晶中间相、用于将蛋白质重构成基于单油酸甘油酯的中间相、用于中间相中蛋白质的功能测定以及用于以手动模式设置结晶的方案。还描述了用于收获微晶的方法。制备负载蛋白质的中间相和手动设置结晶板所需的时间为约1小时。
A detailed protocol for crystallizing membrane proteins that makes use of lipidic mesophases is described. This has variously been referred to as the lipid cubic phase or in meso method. The method has been shown to be quite general in that it has been used to solve X-ray crystallographic structures of prokaryotic and eukaryotic proteins, proteins that are monomeric, homo- and hetero-multimeric, chromophore-containing and chromophore-free, and α-helical and β-barrel proteins. Its most recent successes are the human engineered β2-adrenergic and adenosine A2A G protein-coupled receptors. Protocols are provided for preparing and characterizing the lipidic mesophase, for reconstituting the protein into the monoolein-based mesophase, for functional assay of the protein in the mesophase, and for setting up crystallizations in manual mode. Methods for harvesting micro-crystals are also described. The time required to prepare the protein-loaded mesophase and to set up a crystallization plate manually is about one hour.
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影响因子：
6.1
作者：
Cherezov, V;Caffrey, M
通讯作者：
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影响因子：
3.4
作者：
Cherezov, Vadim;Caffrey, Martin
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Caffrey, Martin
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3.4
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2.9
作者：
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通讯作者：
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影响因子：
6.1
作者：
Cherezov, V;Caffrey, M
通讯作者：
Caffrey, M