Accelerated acquisition of high resolution triple-resonance spectra using non-uniform sampling and maximum entropy reconstruction
Accelerated acquisition of high resolution triple-resonance spectra using non-uniform sampling and maximum entropy reconstruction
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DOI：
10.1016/j.jmr.2004.05.016
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发表时间：
2004-09-01
影响因子：
2.2
通讯作者：
Wagner, G
中科院分区：
文献类型：
--
作者：
Rovnyak, D;Frueh, DP;Wagner, G
非均匀采样显示出在用于获得蛋白质中的H、N、C '、CA和CB核的骨架分配的一套三维NMR实验的获取中提供显著的时间节省：HNCO、HN（CA）CO、HNCA、HN（CO）CA、HNCACB和HN（CO）CACB。非均匀采样意味着根据用户指定的采样时间表，仅在所有递增的演化周期的子集中收集数据。当以1.5mM浓度的膜蛋白的11 kDa胞质结构域的统一方式获得一套六个3D实验时，总共消耗了146小时。采用非均匀采样，在32小时内获得相同的实验，并通过随后的最大熵重建，产生了与通过均匀采集数据的常规傅立叶变换获得的光谱质量相似的光谱。这种方法节省的实验时间可以显着加速通过NMR进行蛋白质结构测定，特别是当与自动分配软件结合使用时，并且能够研究在室温下稳定性差的样品。由于还可以使用节省的时间来获得更多的扫描次数以提高灵敏度，同时保持高分辨率，因此这种方法将有助于扩展NMR研究可获得的蛋白质的大小限制，并为研究具有溶解度问题的样品开辟道路。(C)2004年爱思唯尔公司All rights reserved.
Non-uniform sampling is shown to provide significant time savings in the acquisition of a suite of three-dimensional NMR experiments utilized for obtaining backbone assignments of H, N, C', CA, and CB nuclei in proteins: HNCO, HN(CA)CO, HNCA, HN(CO)CA, HNCACB, and HN(CO)CACB. Non-uniform sampling means that data were collectcd for only a subset of all incremented evolution periods, according to a user-specified sampling schedule. When the suite of six 3D experiments was acquired in a uniform fashion for an 11 kDa cytoplasmic domain of a membrane protein at 1.5 mM concentration, a total of 146 h was consumed. With non-uniform sampling, the same experiments were acquired in 32 h and, through subsequent maximum entropy reconstruction, yielded spectra of similar quality to those obtained by conventional Fourier transform of the uniformly acquired data. The experimental time saved with this methodology can significantly accelerate protein structure determination by NMR, particularly when combined with the use of automated assignment software, and enable the study of samples with poor stability at room temperature. Since it is also possible to use the time savings to acquire a greater numbers of scans to increase sensitivity while maintaining high resolution, this methodology will help extend the size limit of proteins accessible to NMR studies, and open the way to studies of samples that suffer from solubility problems. (C) 2004 Elsevier Inc. All rights reserved.