Smaller Nanodiscs are Suitable for Studying Protein Lipid Interactions by Solution NMR
Smaller Nanodiscs are Suitable for Studying Protein Lipid Interactions by Solution NMR
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DOI：
10.1007/s10930-015-9613-2
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发表时间：
2015-05
期刊：
影响因子：
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通讯作者：
Xiaoming Wang;Zongjun Mu;Yan Li;Y. Bi;Yujuan Wang
中科院分区：
文献类型：
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作者：
Xiaoming Wang;Zongjun Mu;Yan Li;Y. Bi;Yujuan Wang
磷脂双层纳米盘是一种新型的膜模型系统，为膜蛋白的研究提供了“类天然”的膜环境。由膜支架蛋白和磷脂双层组装的纳米盘，具有可以通过改变MSP构建体的氨基酸残基来精确调节的限定尺寸。本文描述了膜支架蛋白缺失突变体ΔMSP的表达和纯化。组装了具有混合脂质的较小纳米盘，并且经分离的31 P NMR光谱显示与具有纯POPC和POPE脂质的纳米盘相同的化学位移，表明它们具有相似的化学环境。将STIM 1-TM掺入纳米盘的成功表明，这种较小的纳米盘系统可用于通过溶液NMR研究膜蛋白。
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 observed31P 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.