Peterson Robert D, Theimer Carla A, Wu Haihong, Feigon Juli
Department of Chemistry and Biochemistry, and Molecular Biology Institute, University of California, Los Angeles, CA 90095-1569, U.S.A.
J Biomol NMR. 2004 Jan;28(1):59-67. doi: 10.1023/B:JNMR.0000012861.95939.05.
NMR spectra of large RNAs are difficult to assign because of extensive spectral overlap and unfavorable relaxation properties. Here we present a new approach to facilitate assignment of RNA spectra using a suite of four 2D-filtered/edited NOESY experiments in combination with base-type-specific isotopically labeled RNA. The filtering method was developed for use in 3D filtered NOESY experiments (Zwahlen et al., 1997), but the 2D versions are both more sensitive and easier to interpret for larger RNAs than their 3D counterparts. These experiments are also useful for identifying intermolecular NOEs in RNA-protein complexes. Applications to NOE assignment of larger RNAs and an RNA-protein complex are presented.
由于广泛的谱线重叠和不利的弛豫特性，大RNA的核磁共振谱很难进行归属。在此，我们提出了一种新方法，通过一套四个二维滤波/编辑的核欧沃豪斯效应谱（NOESY）实验，结合碱基类型特异性同位素标记的RNA，来促进RNA谱的归属。该滤波方法是为三维滤波NOESY实验开发的（茨瓦伦等人，1997年），但二维版本对于大RNA来说比三维版本更灵敏且更易于解读。这些实验对于识别RNA-蛋白质复合物中的分子间核欧沃豪斯效应（NOE）也很有用。本文展示了这些实验在大RNA和RNA-蛋白质复合物的NOE归属中的应用。
