Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR Spectroscopy.
Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR Spectroscopy.
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DOI：
10.1002/anie.201605870
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发表时间：
2016-09-19
影响因子：
16.6
通讯作者：
Kreutz, Christoph
中科院分区：
文献类型：
--
作者：
Juen, Michael Andreas;Wunderlich, Christoph Hermann;Nussbaumer, Felix;Tollinger, Martin;Kontaxis, Georg;Konrat, Robert;Hansen, D. Flemming;Kreutz, Christoph
在这项工作中，一个改进的稳定同位素标记协议的核酸。新的结构单元消除/最小化homomethyl 13 C和1H标量耦合，从而允许质子弛豫分散（RD）实验准确地报告核酸的化学交换。使用位点特异性2 H和13 C标记，将自旋拓扑引入DNA和RNA中，使1H弛豫色散实验以简单的方式适用。新的RNA/DNA结构单元被成功地整合到两个核酸中。A位点RNA先前被证明在微到毫秒的时间范围内经历两个位点的交换过程。使用质子弛豫色散实验的交换参数确定较早可以概括，从而验证了所提出的方法。我们进一步研究了cTAR DNA的动力学，这是一种参与HIV-1病毒复制周期的DNA转录物。同样，交换过程可以被描述和量化。这显示了新标记方案对于核酸的1H RD实验的普遍适用性。
In this work an improved stable isotope labeling protocol for nucleic acids is introduced. The novel building blocks eliminate/minimize homonuclear 13C and 1H scalar couplings thus allowing proton relaxation dispersion (RD) experiments to report accurately on the chemical exchange of nucleic acids. Using site‐specific 2H and 13C labeling, spin topologies are introduced into DNA and RNA that make 1H relaxation dispersion experiments applicable in a straightforward manner. The novel RNA/DNA building blocks were successfully incorporated into two nucleic acids. The A‐site RNA was previously shown to undergo a two site exchange process in the micro‐ to millisecond time regime. Using proton relaxation dispersion experiments the exchange parameters determined earlier could be recapitulated, thus validating the proposed approach. We further investigated the dynamics of the cTAR DNA, a DNA transcript that is involved in the viral replication cycle of HIV‐1. Again, an exchange process could be characterized and quantified. This shows the general applicablility of the novel labeling scheme for 1H RD experiments of nucleic acids.
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影响因子：
3.3
作者：
Frank, Aaron T.;Bae, Sung-Hun;Stelzer, Andrew C.
通讯作者：
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影响因子：
16.6
作者：
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15
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影响因子：
64.8
作者：
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通讯作者：
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