Wang Xiaoming, Mu Zongjun, Li Yan, Bi Yunchen, Wang Yujuan
High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Science, Hefei, 230031, Anhui, China.
Protein J. 2015 Jun;34(3):205-11. doi: 10.1007/s10930-015-9613-2.
Phospholipid bilayer nanodiscs, a newly developed model membrane system, provides "native-like" membrane environment for membrane protein studies. Nanodiscs assembled by membrane scaffold protein and phospholipid bilayer, with defined sizes that can be accurately regulated by changing the amino acid residues of the MSP construct. Herein we described the expression and purification of ΔMSP, a deletion mutant of the membrane scaffold protein. Smaller nanodiscs with mixed lipids were assembled, and the observed (31)P NMR spectra showed identical chemical shifts to those of nanodiscs with pure POPC and POPE lipids, indicating they share similar chemical environments. The success of incorporation STIM1-TM into nanodiscs indicated the application of this smaller nanodisc system can be used to membrane protein studies by solution NMR.
磷脂双层纳米盘是一种新开发的模型膜系统，为膜蛋白研究提供了“类天然”的膜环境。纳米盘由膜支架蛋白和磷脂双层组装而成，其大小可通过改变MSP构建体的氨基酸残基精确调控。在此，我们描述了膜支架蛋白的缺失突变体ΔMSP的表达和纯化。组装了具有混合脂质的较小纳米盘，观察到的（31）P NMR光谱显示其化学位移与纯POPC和POPE脂质的纳米盘相同，表明它们具有相似的化学环境。将STIM1-TM整合到纳米盘中的成功表明，这种较小的纳米盘系统可应用于通过溶液核磁共振进行膜蛋白研究。
