Sensitivity and resolution enhanced solid-state NMR for paramagnetic systems and biomolecules under very fast magic angle spinning.
Sensitivity and resolution enhanced solid-state NMR for paramagnetic systems and biomolecules under very fast magic angle spinning.
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DOI：
10.1021/ar4000482
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发表时间：
2013-09-17
影响因子：
18.3
通讯作者：
Ishii, Yoshitaka
中科院分区：
文献类型：
--
作者：
Parthasarathy, Sudhakar;Nishiyama, Yusuke;Ishii, Yoshitaka
最近的研究在快速魔角旋转（MAS）的方法已经大大提高了分辨率和灵敏度的NMR光谱的生物分子和固体材料。在这个帐户中，我们总结了最近和正在进行的发展，在这一领域提出13 C和1H固态NMR（SSNMR）的研究顺磁系统和生物分子下快速MAS从我们的实验室。首先，我们描述了如何非常快的MAS（VFMAS）在20 kHz的旋转速度使我们能够克服主要困难，在1H和13 C高分辨率SSNMR的顺磁系统。因此，我们可以将灵敏度和分辨率提高几个数量级。使用快速回收（~ms/扫描）和短的1H T1值，我们可以在μg尺度上对顺磁性体系进行1H SSNMR微观分析，其灵敏度大大提高。其次，我们讨论了如何在旋转速度大于~40 kHz的VFMAS可以提高13 C生物分子SSNMR测量的灵敏度和分辨率。VFMAS下的低功率1H去耦方案通过~10 kHz的标称1H RF辐照为13 C SSNMR提供了优异的光谱分辨率。通过结合VFMAS方法和通过顺磁掺杂增强的1H T1弛豫，我们可以在现代生物分子SSNMR实验中实现极快的再循环。13 C标记的泛素掺杂10 mM Cu-EDTA的实验证明了这种新方法，称为顺磁辅助浓缩数据收集（PACC），提高了灵敏度。最后，我们研究13 C SSNMR测量生物分子在更快的MAS在更高的领域。我们在750-800 MHz的1H NMR频率下获得的Aβ淀粉样蛋白原纤维和GB 1微晶的初步13 C SSNMR数据表明，PACC方法和超高场的组合使用可以允许在50-200 nmol水平上对蛋白质进行常规多维SSNMR分析。最后，我们简要地讨论了在~100 kHz的旋转速度下，在超快MAS下，用13 C SSNMR研究双分子的前景。
Recent research in fast magic angle spinning (MAS) methods has drastically improved in the resolution and sensitivity for NMR spectroscopy of biomolecules and materials in solids. In this Account, we summarizes recent and ongoing developments in this area by presenting 13C and 1H solid-state NMR (SSNMR) studies on paramagnetic systems and biomolecules under fast MAS from our laboratories. First, we describe how very fast MAS (VFMAS) at the spinning speed of 20 kHz allows us to overcome major difficulties in 1H and 13C high-resolution SSNMR of paramagnetic systems. As a result, we can enhance both sensitivity and resolution by up to a few orders of magnitude. Using fast recycling (~ms/scan) using short 1H T1 values we can perform 1H SSNMR micro-analysis of paramagnetic systems in the μg scale with greatly improved sensitivity over that for diamagnetic systems. Second, we discuss how VFMAS at a spinning speed greater than ~40 kHz can enhance the sensitivity and resolution of 13C biomolecular SSNMR measurements. Low-power 1H decoupling schemes under VFMAS offer excellent spectral resolution for 13C SSNMR by nominal 1H RF irradiation at ~10 kHz. By combining the VFMAS approach and enhanced 1H T1 relaxation by paramagnetic doping we can achieve extremely fast recycling in modern biomolecular SSNMR experiments. Experiments for 13C-labeled ubiquitin doped with 10 mM Cu-EDTA demonstrate how effectively this new approach, called paramagnetic assisted condensed data collection (PACC), enhances the sensitivity. Lastly, we examine 13C SSNMR measurements for biomolecules under faster MAS at a higher field. Our preliminary 13C SSNMR data of Aβ amyloid fibrils and GB1 microcrystals acquired at 1H NMR frequencies of 750-800 MHz suggest that the combined use of the PACC approach and the ultra-high fields could allow for routine multi-dimensional SSNMR analyses of proteins at the 50-200 nmol level. Also, we briefly discuss the prospects for studying bimolecules using 13C SSNMR under ultra fast MAS at the spinning speed of ~100 kHz.
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影响因子：
4.4
作者：
Ishii, Y
通讯作者：
Ishii, Y
DOI：
10.1073/pnas.0708460105
发表时间：
2008-11-11
影响因子：
11.1
作者：
Balayssac, Stephane;Bertini, Ivano;Luchinat, Claudio
通讯作者：
Luchinat, Claudio
影响因子：
2.2
作者：
Ishii, Y;Tycko, R
通讯作者：
Tycko, R
影响因子：
15
作者：
Ishii, Y;Wickramasinghe, NP;Chimon, S
通讯作者：
Chimon, S
影响因子：
3.2
作者：
Jaroniec, Christopher P.
通讯作者：
Jaroniec, Christopher P.