Detecting ligand binding to a small RNA target via saturation transfer difference NMR experiments in D2O and H2O
Detecting ligand binding to a small RNA target via saturation transfer difference NMR experiments in D2O and H2O
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DOI：
10.1021/ja027526z
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发表时间：
2002-11-13
影响因子：
15
通讯作者：
James, TL
中科院分区：
文献类型：
--
作者：
Mayer, M;James, TL
小RNA基序被用作通过饱和转移差（STD）NMR实验进行基于配体的NMR筛选的靶标。在STD实验中使用小RNA靶的先决条件，如饱和时间，频率和脉冲，进行了讨论。我们还表明，由于D_2O中R_1弛豫速率降低，所以用D_2O代替H_2O作溶剂是有利的。已知核糖体A位点的27个核苷酸模型以高亲和力结合氨基糖苷类巴龙霉素。这种结合相互作用可以很容易地检测到，证明了STD NMR实验作为RNA-配体相互作用筛选工具的有效性。
A small RNA motif is used as a target for ligand-based NMR-screening by saturation transfer difference (STD) NMR experiments. The prerequisites for using a small RNA target in STD experiments, such as saturation time, frequency, and pulses, are discussed. We also show that it is of advantage to use D2O as solvent instead of H2O due to the reducedR1relaxation rate in D2O. The 27-nucleotide model of the ribosomal A-site was known to bind the aminoglycoside paromomycin with high affinity. This binding interaction could be detected easily, proving the effectiveness of STD NMR experiments as a screening tool for RNA−ligand interactions.