Andréll Juni, Tate Christopher G
MRC Laboratory of Molecular Biology, Cambridge, UK.
Mol Membr Biol. 2013 Feb;30(1):52-63. doi: 10.3109/09687688.2012.703703. Epub 2012 Sep 11.
The number of structures of integral membrane proteins from higher eukaryotes is steadily increasing due to a number of innovative protein engineering and crystallization strategies devised over the last few years. However, it is sobering to reflect that these structures represent only a tiny proportion of the total number of membrane proteins encoded by a mammalian genome. In addition, the structures determined to date are of the most tractable membrane proteins, i.e., those that are expressed functionally and to high levels in yeast or in insect cells using the baculovirus expression system. However, some membrane proteins that are expressed inefficiently in these systems can be produced at sufficiently high levels in mammalian cells to allow structure determination. Mammalian expression systems are an under-used resource in structural biology and represent an effective way to produce fully functional membrane proteins for structural studies. This review will discuss examples of vertebrate membrane protein overexpression in mammalian cells using a variety of viral, constitutive or inducible expression systems.
由于过去几年设计出的一些创新型蛋白质工程和结晶策略，来自高等真核生物的整合膜蛋白结构数量正在稳步增加。然而，令人清醒的是，这些结构仅占哺乳动物基因组编码的膜蛋白总数的极小一部分。此外，迄今为止确定的结构是最易处理的膜蛋白，即那些使用杆状病毒表达系统在酵母或昆虫细胞中功能性表达且表达水平较高的膜蛋白。然而，一些在这些系统中表达效率低下的膜蛋白可以在哺乳动物细胞中以足够高的水平产生，从而进行结构测定。哺乳动物表达系统在结构生物学中是一种未充分利用的资源，是为结构研究生产功能完全正常的膜蛋白的有效途径。本综述将讨论使用多种病毒、组成型或诱导型表达系统在哺乳动物细胞中过表达脊椎动物膜蛋白的实例。
