Longitudinal-Relaxation-Enhanced NMR Experiments for the Study of Nucleic Acids in Solution

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• 作者：Jonathan Farjon ; Jrme Boisbouvier ; Paul Schanda ; Arthur Pardi ; Jean-Pierre Simorre ; Bernhard Brutscher
• 刊名：Journal of the American Chemical Society
• 出版年：2009
• 出版时间：June 24, 2009
• 年：2009
• 卷：131
• 期：24
• 页码：8571-8577
• 全文大小：323K
• 年卷期：v.131,no.24(June 24, 2009)
• ISSN：1520-5126

文摘

Atomic-resolution information on the structure and dynamics of nucleic acids is essential for a better understanding of the mechanistic basis of many cellular processes. NMR spectroscopy is a powerful method for studying the structure and dynamics of nucleic acids; however, solution NMR studies are currently limited to relatively small nucleic acids at high concentrations. Thus, technological and methodological improvements that increase the experimental sensitivity and spectral resolution of NMR spectroscopy are required for studies of larger nucleic acids or protein−nucleic acid complexes. Here we introduce a series of imino-proton-detected NMR experiments that yield an over 2-fold increase in sensitivity compared to conventional pulse schemes. These methods can be applied to the detection of base pair interactions, RNA−ligand titration experiments, measurement of residual dipolar ¹⁵N−¹H couplings, and direct measurements of conformational transitions. These NMR experiments employ longitudinal spin relaxation enhancement techniques that have proven useful in protein NMR spectroscopy. The performance of these new experiments is demonstrated for a 10 kDa TAR-TAR*_GA RNA kissing complex and a 26 kDa tRNA.