Labeling strategies for 13C-detected aligned-sample solid-state NMR of proteins
Labeling strategies for 13C-detected aligned-sample solid-state NMR of proteins
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DOI：
10.1016/j.jmr.2009.08.012
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发表时间：
2009-12-01
影响因子：
2.2
通讯作者：
Opella, Stanley J.
中科院分区：
文献类型：
--
作者：
Filipp, Fabian V.;Sinha, Neeraj;Opella, Stanley J.
C-13检测的固态核磁共振实验比相应的n -15检测的实验具有更高的灵敏度，石油固定，同位素标记的蛋白质排列样品。描述了几种裁剪同位素标记的方法，这些方法导致空间隔离的13C位点，从而使C-13之间的偶极子-偶极子耦合最小化，从而在使用C-13检测的一维或多维NMR实验中，在间接或直接维度上消除了对同核C-13-C-13解耦的需要。随机分数C-13标记的最佳百分比在25%到35%之间。特定标记的甘油和葡萄糖可以在碳源上使用，以定制同位素标记，选择取决于对特定研究感兴趣的共振。对于蛋白质骨架的研究，发现细菌在含[2-C-13]-葡萄糖的培养基上生长最有效。(C) 2009爱思唯尔公司版权所有。
C-13 detected solid-state NMR experiments have substantially higher sensitivity than the corresponding N-15-detected experiments oil stationary, aligned samples of isotopically labeled proteins. Several methods for tailoring the isotopic labeling are described that result in spatially isolated 13C sites so that dipole-dipole couplings among the C-13 are minimized, thus eliminating the need for homonuclear C-13-C-13 decoupling in either indirect or direct dimensions of one- OF multi-dimensional NMR experiments that employ C-13 detection. The optimal percentage for random fractional C-13 labeling is between 25% and 35%. Specifically labeled glycerol and glucose can be used at the carbon Sources to tailor the isotopic labeling, and the choice depends on the resonances of interest for a particular study. For investigations of the protein backbone, growth of the bacteria on [2-C-13]-glucose-containing media was found to be most effective. (C) 2009 Elsevier Inc. All rights reserved.