Tate C G
MRC Laboratory of Molecular Biology, Hills Road, CB2 2QH, Cambridge, UK.
FEBS Lett. 2001 Aug 31;504(3):94-8. doi: 10.1016/s0014-5793(01)02711-9.
Recent successes in the determination of atomic resolution structures of integral membrane proteins have relied on purifying the proteins from abundant natural sources. In contrast, the majority of mammalian receptors, ion channels and transporters need to be overexpressed to obtain sufficient material for structural studies. This has often proved to be very difficult. Overexpression studies on a wide range of mammalian membrane proteins have shown that a few can be expressed functionally in bacteria, but many others require an insect or mammalian cell host for activity or high level expression. The serotonin transporter, which has been expressed in all the major hosts available, is a good example that has given insights into the problem of overexpressing mammalian membrane proteins for structural studies.
近期在解析整合膜蛋白原子分辨率结构方面取得的成功,依赖于从丰富的天然来源中纯化蛋白质。相比之下,大多数哺乳动物受体、离子通道和转运蛋白需要进行过表达,以获得足够的材料用于结构研究。事实证明,这往往非常困难。对多种哺乳动物膜蛋白的过表达研究表明,少数蛋白可以在细菌中功能性表达,但许多其他蛋白需要昆虫或哺乳动物细胞宿主才能实现活性或高水平表达。血清素转运蛋白已在所有可用的主要宿主中进行了表达,它就是一个很好的例子,为在结构研究中过表达哺乳动物膜蛋白的问题提供了见解。