Tate Christopher G
MRC Laboratory of Molecular Biology, Cambridge, UK.
Methods Mol Biol. 2010;601:187-203. doi: 10.1007/978-1-60761-344-2_12.
Crystallisation of integral membranes requires milligrams of purified protein in a homogeneous, monodisperse state, and crucially, the membrane protein must also be fully functional and stable. The stability of membrane proteins in solution is dependent on the type of detergents used, but unfortunately the use of the most stabilising detergent can often decrease the probability of obtaining crystals that diffract to high resolution, especially of small membrane proteins. A number of strategies have been developed to facilitate the purification of membrane proteins in a functional form, which have led to new possibilities for structure determination.
完整膜蛋白的结晶需要数毫克处于均匀、单分散状态的纯化蛋白,关键的是,膜蛋白还必须具备完全的功能且稳定。膜蛋白在溶液中的稳定性取决于所用去污剂的类型,但遗憾的是,使用最具稳定作用的去污剂往往会降低获得能产生高分辨率衍射晶体的概率,对于小的膜蛋白尤其如此。人们已开发出多种策略来促进以功能形式纯化膜蛋白,这为结构测定带来了新的可能性。