2'-hydroxyl proton positions in helical RNA from simultaneously measured heteronuclear scalar couplings and NOEs.
2'-hydroxyl proton positions in helical RNA from simultaneously measured heteronuclear scalar couplings and NOEs.
复制标题
来自同时测量的异核标量耦合和 NOE 的螺旋 RNA 中的 2-羟基质子位置。
DOI：
10.1021/ja0606226
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发表时间：
2006
影响因子：
15
通讯作者：
Bax,Ad
中科院分区：
文献类型：
--
作者：
Ying,Jinfa;Bax,Ad
RNA中的2 '-羟基起着重要的结构作用;它定义了小沟中的水合作用，影响RNA的热力学稳定性，并经常参与RNA催化。为了更好地研究RNA中的这个重要官能团，我们描述了一个恒时HMQC−IPAP−NOESY 3D NMR实验。它同时产生从核糖质子到2 'OH质子的高分辨13 C分离的NOE，以及JC-OH偶联的E. COSY型测量，从而允许定量研究RNA中2' OH质子的取向。观察到的NOE模式表明A型螺旋RNA中的2 'OH键主要朝向碱基结构域，这进一步得到小的（1.3 ± 0.7 Hz）3 JC 1'-2 'OH和相对大的（4.2 ± 0.7 Hz）3 JC 3'-2 'OH偶联的支持。恒时HMQC−IPAP−NOESY适用于测量蛋白质和核酸中快速交换质子的相互作用。
The 2‘-hydroxyl group in RNA plays an important structural role; it defines hydration in the minor groove, impacts thermodynamic stability of RNA, and often participates in RNA catalysis. To better study this important functional group in RNA, we describe a constant-time HMQC−IPAP−NOESY 3D NMR experiment. It simultaneously yields highly resolved13C-separated NOEs from ribose protons to the 2‘OH proton, as well as E.COSY-type measurement ofJC-OHcouplings, thereby permitting a quantitative study of the orientation of the 2‘OH proton in RNA. The observed NOE patterns indicate that the 2‘OH bonds in A-form helical RNA are primarily oriented toward the base domain, as further supported by small (1.3 ± 0.7 Hz)3JC1‘-2‘OHand relatively large (4.2 ± 0.7 Hz)3JC3‘-2‘OHcouplings. The constant-time HMQC−IPAP−NOESY is suitable for measurement of interactions of rapidly exchanging protons in proteins and nucleic acids.