A high-resolution solid-state NMR approach for the structural studies of bicelles
A high-resolution solid-state NMR approach for the structural studies of bicelles
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DOI：
10.1021/ja061153a
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发表时间：
2006-05-17
影响因子：
15
通讯作者：
Ramamoorthy, Ayyalusamy
中科院分区：
文献类型：
--
作者：
Dvinskikh, Sergey;Durr, Ulrich;Ramamoorthy, Ayyalusamy
双胞越来越多地被用作核磁共振实验中的膜模拟系统，以研究膜蛋白的结构。在这项研究中，我们证明了2D固态NMR方法的有效性，该方法可用于测量结构约束，如1H，13 C和31 P核之间的异源偶极耦合，而不需要同位素富集。该方法不像目前使用的双帧分离局部场（SLF）技术（诸如PISEMA）那样需要高射频功率。此外，可以精确地测量多个偶极耦合，并且强偶极耦合的存在不会抑制弱耦合。从磁性排列的DMPC：DHPC双胞获得的高分辨率光谱，即使在存在肽的情况下，也表明这种方法将有助于理解脂质-蛋白质相互作用，这种相互作用在形成膜蛋白的功能中起着至关重要的作用。
Bicelles are increasingly being used as membrane mimicking systems in NMR experiments to investigate the structure of membrane proteins. In this study, we demonstrate the effectiveness of a 2D solid-state NMR approach that can be used to measure the structural constraints, such as heteronuclear dipolar couplings between1H,13C, and31P nuclei, in bicelles without the need for isotopic enrichment. This method does not require a high radio frequency power unlike the presently used rotating-frame separated-local-field (SLF) techniques, such as PISEMA. In addition, multiple dipolar couplings can be measured accurately, and the presence of a strong dipolar coupling does not suppress the weak couplings. High-resolution spectra obtained from magnetically aligned DMPC:DHPC bicelles even in the presence of peptides suggest that this approach will be useful in understanding lipid−protein interactions that play a vital role in shaping up the function of membrane proteins.