通过使用灵敏的 13C-甲基标记物，通过纵向交换和 CPMG 弛豫色散 NMR 光谱学探测 RNA 动力学。
Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive 13C-methyl label.
机构信息
Institute of Organic Chemistry, Leopold Franzens University, Innrain 52a, 6020 Innsbruck, Austria.
出版信息
Nucleic Acids Res. 2011 May;39(10):4340-51. doi: 10.1093/nar/gkq1361. Epub 2011 Jan 19.
The refolding kinetics of bistable RNA sequences were studied in unperturbed equilibrium via (13)C exchange NMR spectroscopy. For this purpose a straightforward labeling technique was elaborated using a 2'-(13)C-methoxy uridine modification, which was prepared by a two-step synthesis and introduced into RNA using standard protocols. Using (13)C longitudinal exchange NMR spectroscopy the refolding kinetics of a 20 nt bistable RNA were characterized at temperatures between 298 and 310K, yielding the enthalpy and entropy differences between the conformers at equilibrium and the activation energy of the refolding process. The kinetics of a more stable 32 nt bistable RNA could be analyzed by the same approach at elevated temperatures, i.e. at 314 and 316 K. Finally, the dynamics of a multi-stable RNA able to fold into two hairpin- and a pseudo-knotted conformation was studied by (13)C relaxation dispersion NMR spectroscopy.
通过（13）C 交换 NMR 光谱法研究了未受干扰平衡中双稳 RNA 序列的重折叠动力学。为此，通过使用 2' -（13）C-甲氧基尿嘧啶修饰的简单标记技术进行了阐述，该修饰通过两步合成制备，并使用标准方案引入 RNA。使用（13）C 纵向交换 NMR 光谱法，在 298 至 310K 之间的温度下对 20nt 双稳 RNA 的重折叠动力学进行了表征，得到了平衡构象之间的焓和熵差以及重折叠过程的活化能。通过相同的方法可以在较高温度下（即 314 和 316K）分析更稳定的 32nt 双稳 RNA 的动力学。最后，通过（13）C 弛豫分散 NMR 光谱法研究了能够折叠成两个发夹和一个拟环结构的多稳 RNA 的动力学。
Zotero 插件