Base-type-selective high-resolution 13C edited NOESY for sequential assignment of large RNAs
Base-type-selective high-resolution 13C edited NOESY for sequential assignment of large RNAs
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DOI：
10.1023/a:1008340210079
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发表时间：
2001-02-01
影响因子：
2.7
通讯作者：
Simorre, JP
中科院分区：
文献类型：
--
作者：
Brutscher, B;Boisbouvier, J;Simorre, JP
大量的光谱重叠是大RNA核磁共振研究的一个主要问题。在这里，我们提出了NMR技术的分辨率增强和光谱简化完全C-13标记的RNA。将TROSY型实验与多带选择性homopolymer C-13去偶相结合，获得了高分辨率的H-1-C-13相关谱。额外的C-C滤波器序列执行碱基类型选择性光谱编辑。由于TROSY型自旋演化，滤波期间的信号损失显著减少。这些工具可以插入任何C-13编辑的多维NMR实验中。作为一个例子，我们选择了C-13编辑的NOESY，这是RNA序列共振分配的关键实验。应用于33个核苷酸的RNA适体和76个核苷酸的tRNA说明了这种新方法的潜力。
Extensive spectral overlap presents a major problem for the NMR study of large RNAs. Here we present NMR techniques for resolution enhancement and spectral simplification of fully C-13 labelled RNA. High-resolution H-1-C-13 correlation spectra are obtained by combining TROSY-type experiments with multiple-band-selective homonuclear C-13 decoupling. An additional C-C filter sequence performs base-type-selective spectral editing. Signal loss during the filter is significantly reduced because of TROSY-type spin evolution. These tools can be inserted in any C-13-edited multidimensional NMR experiment. As an example we have chosen the C-13-edited NOESY which is a crucial experiment for sequential resonance assignment of RNA. Application to a 33-nucleotide RNA aptamer and a 76-nucleotide tRNA illustrates the potential of this new methodology.