3D DUMAS: Simultaneous acquisition of three-dimensional magic angle spinning solid-state NMR experiments of proteins
3D DUMAS: Simultaneous acquisition of three-dimensional magic angle spinning solid-state NMR experiments of proteins
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DOI：
10.1016/j.jmr.2012.04.006
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发表时间：
2012-07-01
影响因子：
2.2
通讯作者：
Veglia, Gianluigi
中科院分区：
文献类型：
--
作者：
Gopinath, T.;Veglia, Gianluigi
使用大仲马（双采集魔角旋转）固态NMR方法，我们创建了新的脉冲方案，能够同时采集均匀C-13，N-15标记蛋白质的三维（3D）实验。这些新实验利用从H-1到C-13和N-15的同时交叉极化（SIM-CP）来同时获得两个3D实验。这是通过C-13和N-15之间的双向极化转移以及固态NMR中的长寿命N-15 z-极化而实现的。为了证明这种方法的强大功能，将四个3D脉冲序列（NCACX、CANCO、NCOCX、CON（CA）CX）组合成两个脉冲序列（3D DUMAS-NCACX-CANCO、3D DUMAS-NCOCX-CON（CA）CX），允许同时采集这些实验，将实验时间缩短约一半。重要的是，仅3D DUMAS-NCACX-CANCO实验就可以获得微晶U-C-13，N-15泛素的大部分骨架顺序共振归属。大仲马方法是通用的，适用于许多3D实验，几乎是NMR光谱仪性能的两倍。(C)2012 Elsevier Inc. All rights reserved.
Using the DUMAS (Dual acquisition Magic Angle Spinning) solid-state NMR approach, we created new pulse schemes that enable the simultaneous acquisition of three dimensional (3D) experiments on uniformly C-13, N-15 labeled proteins. These new experiments exploit the simultaneous cross-polarization (SIM-CP) from H-1 to C-13 and N-15 to acquire two 3D experiments simultaneously. This is made possible by bidirectional polarization transfer between C-13 and N-15 and the long living N-15 z-polarization in solid state NMR. To demonstrate the power of this approach, four 3D pulse sequences (NCACX, CANCO, NCOCX, CON(CA)CX) are combined into two pulse sequences (3D DUMAS-NCACX-CANCO, 3D DUMAS-NCOCX-CON(CA)CX) that allow simultaneous acquisition of these experiments, reducing the experimental time by approximately half. Importantly, the 3D DUMAS-NCACX-CANCO experiment alone makes it possible to obtain the majority of the backbone sequential resonance assignments for microcrystalline U-C-13,N-15 ubiquitin. The DUMAS approach is general and applicable to many 3D experiments, nearly doubling the performance of NMR spectrometers. (C) 2012 Elsevier Inc. All rights reserved.