F1F2-selective NMR spectroscopy
F1F2-selective NMR spectroscopy
复制标题
F1F2-选择性核磁共振波谱
DOI：
10.1007/s10858-017-0113-x
复制
发表时间：
2017
影响因子：
2.7
通讯作者：
Sugase Kenji
中科院分区：
文献类型：
--
作者：
Walinda Erik;Morimoto Daichi;Shirakawa Masahiro;Sugase Kenji
傅里叶变换核磁共振光谱学为蛋白质和核酸的结构、相互作用和动态运动提供了前所未有的洞察力。传统的生物分子NMR依赖于三维和四维（4D）数据矩阵的采集来分别建立频域F1、F2、F3和F1、F2、F3、F4中的化学位移之间的相关性。虽然信息丰富，但这些数据集需要大量的采集时间，视觉上非常不直观，需要专业知识来处理，并对频域的暗区和亮区进行同等采样。在这里，我们提出了另一种方法来获得多维化学位移相关的生物分子。这种策略集中在一个狭窄的频率范围，F1 F2，在同一时间和记录的resultingF 3F 4相关谱的二维NMR。因此，仅对包含F1 F2中的信号的频域区域（“亮区域”）进行采样。F1 F2选择通过使用弱射频场的Hartmann-Hahn交叉极化来实现。这种方法揭示的信息相当于传统的4D实验，而降维可以缩短总的采集时间，简化光谱处理，解释和比较分析。通过泛素和脂肪酸结合蛋白4（FABP 4）的从头分配、结构和动力学研究，说明了F1 F2选择性方法的潜在适用性。这一概念的进一步扩展可能会产生新的选择性核磁共振实验，以帮助研究位点特异性结构动力学、蛋白质-蛋白质相互作用和蛋白质结构的变构调节。
Fourier transform NMR spectroscopy has provided unprecedented insight into the structure, interaction and dynamic motion of proteins and nucleic acids. Conventional biomolecular NMR relies on the acquisition of three-dimensional and four-dimensional (4D) data matrices to establish correlations between chemical shifts in the frequency domainsF1,F2,F3andF1,F2,F3,F4respectively. While rich in information, these datasets require a substantial amount of acquisition time, are visually highly unintuitive, require expert knowledge to process, and sample dark and bright regions of the frequency domains equally. Here, we present an alternative approach to obtain multidimensional chemical shift correlations for biomolecules. This strategy focuses on one narrow frequency range,F1F2, at a time and records the resultingF3F4correlation spectrum by two-dimensional NMR. As a result, only regions of the frequency domain that contain signals inF1F2(“bright regions”) are sampled.F1F2selection is achieved by Hartmann–Hahn cross-polarization using weak radio frequency fields. This approach reveals information equivalent to that of a conventional 4D experiment, while the dimensional reduction may shorten the total acquisition time and simplifies spectral processing, interpretation and comparative analysis. Potential applicability of theF1F2-selective approach is illustrated by de novo assignment, structural and dynamics studies of ubiquitin and fatty-acid binding protein 4 (FABP4). Further extension of this concept may spawn new selective NMR experiments to aid studies of site-specific structural dynamics, protein–protein interactions and allosteric modulation of protein structure.