Structural analysis of metal ion ligation to nucleotides and nucleic acids using pulsed EPR spectroscopy.
Structural analysis of metal ion ligation to nucleotides and nucleic acids using pulsed EPR spectroscopy.
复制标题
使用脉冲 EPR 光谱法对金属离子与核苷酸和核酸的连接进行结构分析。
DOI：
10.1021/ja0112238
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发表时间：
2002
影响因子：
15
通讯作者：
Britt,RDavid
中科院分区：
文献类型：
--
作者：
Hoogstraten,CharlesG;Grant,ChristopherV;Horton,ThomasE;DeRose,VictoriaJ;Britt,RDavid
Metal ions play key structural and functional roles in many nucleic acid systems, particularly as required cofactors for many catalytic RNA molecules (ribozymes). We apply the pulsed EPR technologies of electron spin−echo envelope modulation and electron spin−echo-electron nuclear double resonance to the structural analysis of the paramagnetic metal ion Mn(II) bound to nucleotides and nucleic acids. We demonstrate that pulsed EPR, supplemented with specific isotope labeling, can characterize ligation to nucleotide base nitrogens, outer-sphere interactions with phosphate groups, distances to sites of specific2H atom labels, and the hydration level of the metal ion. These techniques allow a comprehensive structural analysis of the mononucleotide model system MnGMP. Spectra of phenylalanine-specific transfer RNA from budding yeast and of the hammerhead ribozyme demonstrate the applicability of the methods to larger, structured RNA systems. This suite of experiments opens the way to detailed structural characterization of specifically bound metal ions in a variety of ribozymes and other nucleic acids of biological interest.
