Nollert Peter
deCODE genetics, BioStructures Group, 7869 NE Day RdW, Bainbridge Island, WA 98110, USA.
Methods. 2004 Nov;34(3):348-53. doi: 10.1016/j.ymeth.2004.03.030.
This review provides detailed procedures for the crystallization of membrane proteins via the lipidic cubic phase method. Bacteriorhodopsin-specific, hands-on protocols are given for (i) the preparation of bacteriohordopsin from purple membrane by monomerization in octylglucoside and gel filtration chromatography or by selective extraction after pre-treatment with dodecyl-trimethylammonium bromide, (ii) the incorporation of bacteriorhodopsin into lipidic cubic phases by mixing in vials or within coupled syringes and, (iii) the crystallization of bacteriorhodopsin in the lipidic matrix by adding a solid salt or an overlaying with a solution. References for further useful procedures and materials are listed in order to provide biochemists and crystallographers with all information that is necessary to grow crystals of the membrane protein bacteriorhodopsin.
本综述提供了通过脂质立方相法结晶膜蛋白的详细步骤。给出了针对细菌视紫红质的实际操作方案，包括：（i）通过在辛基葡糖苷中单体化并经凝胶过滤色谱法从紫膜制备细菌视紫红质，或在用十二烷基三甲基溴化铵预处理后进行选择性提取；（ii）通过在小瓶中混合或在连接的注射器内将细菌视紫红质掺入脂质立方相；以及（iii）通过添加固体盐或覆盖溶液在脂质基质中使细菌视紫红质结晶。列出了进一步有用的操作步骤和材料的参考文献，以便为生物化学家和晶体学家提供生长膜蛋白细菌视紫红质晶体所需的所有信息。
