Detergent optimized membrane protein reconstitution in liposomes for solid state NMR.
Detergent optimized membrane protein reconstitution in liposomes for solid state NMR.
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用于固态 NMR 的脂质体中去垢剂优化的膜蛋白重构。
DOI：
10.1021/bi500144h
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发表时间：
2014-04-22
期刊：
影响因子：
2.9
通讯作者：
Cross TA
中科院分区：
文献类型：
--
作者：
Murray DT;Griffin J;Cross TA
对于小螺旋膜蛋白，其结构对其环境高度敏感，固态核磁共振是一种结构技术，可以表征这些膜蛋白在类天然脂质双层和蛋白脂质体中的特征。迄今为止，文献中尚未提出一种系统方法来评估增溶去污剂对膜蛋白固态核磁共振的蛋白脂质体制剂的影响。提出了一组实验，旨在确定最适合透析介导的重构样品制备的条件。使用结核分枝杆菌的膜蛋白来说明该方法。结果表明，稳定大部分蛋白质的去污剂并不总是理想的，有时无法通过透析去除。通过关注去垢剂的脂质和蛋白质结合特性，可以轻松生产蛋白脂质体制剂，为定向样品和魔角旋转固态 NMR 提供双倍的信噪比。该方法将允许在脂质双层环境中对更多膜蛋白药物靶标进行结构表征。
For small helical membrane proteins, their structures are highly sensitive to their environment, and solid state NMR is a structural technique that can characterize these membrane proteins in native-like lipid bilayers and proteoliposomes. To date, a systematic method by which to evaluate the effect of the solubilizing detergent on proteoliposome preparations for solid state NMR of membrane proteins has not been presented in the literature. A set of experiments are presented aimed at determining the conditions most amenable to dialysis mediated reconstitution sample preparation. A membrane protein from M. tuberculosis is used to illustrate the method. The results show that a detergent that stabilizes the most protein is not always ideal and sometimes cannot be removed by dialysis. By focusing on the lipid and protein binding properties of the detergent, proteoliposome preparations can be readily produced, which provide double the signal-to-noise ratios for both the oriented sample and magic angle spinning solid state NMR. The method will allow more membrane protein drug targets to be structurally characterized in lipid bilayer environments.
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影响因子：
14.8
作者：
通讯作者：
--
影响因子：
56.9
作者：
KETCHEM, RR;HU, W;CROSS, TA
通讯作者：
CROSS, TA
影响因子：
3
作者：
Hanford, M;Peeples, TL
通讯作者：
Peeples, TL
DOI：
10.1016/j.bbagen.2013.02.001
发表时间：
2013-06-01
影响因子：
3
作者：
Low, Christian;Moberg, Per;Nordlund, Par
通讯作者：
Nordlund, Par
影响因子：
2.2
作者：
McNeill, Seth A.;Gor'kov, Peter L.;Long, Joanna R.
通讯作者：
Long, Joanna R.