Synthesis of 5-fluoropyrimidine Nucleotides as sensitive NMR probes of RNA structure
Synthesis of 5-fluoropyrimidine Nucleotides as sensitive NMR probes of RNA structure
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DOI：
10.1021/ja073825i
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发表时间：
2007-12-05
影响因子：
15
通讯作者：
Williamson, James R.
中科院分区：
文献类型：
--
作者：
Hennig, Mirko;Scott, Lincoln G.;Williamson, James R.
氟化 5'-三磷酸类似物 5F-UTP 和 5F-CTP 的酶促合成方法已被开发出来，以促进 RNA 的 F-19 标记用于生物物理研究。 HIV-2 TAR RNA 使用这些类似物通过 T7 RNA 聚合酶的体外转录反应合成。将 5F-U 或 5F-C 类似物均匀掺入 HIV-2 TAR RNA 转录本不会显着改变 RNA 结构或热力学稳定性。介绍并讨论了提供选择性距离信息的氟观察同核 F-19-F-19 和异核 F-19-H-1 NOE 实验。有效合成 5F-UTP 以及首次 5F-CTP 的可用性将有助于利用 F-19 标记的优势，在 RNA 的结构、配体结合和动态研究中使用 5F 标记的 RNA。
Enzymatic synthesis methods for the fluorinated 5'-triphosphqte analogues 5F-UTP and 5F-CTP have been developed to facilitate F-19-labeling of RNAs for biophysical studies. HIV-2 TAR RNAs were synthesized using these analogues by in vitro transcription reactions using T7 RNA polymerase. The uniform incorporation of 5F-U or 5F-C analogues into HIV-2 TAR RNA transcripts does not significantly alter the RNA structure or thermodynamic stability. Fluorine observed homonuclear F-19-F-19 and heteronuclear F-19-H-1 NOE experiments providing selective distance information are presented and discussed. The availability of efficient synthesis of 5F-UTP, and for the first time, 5F-CTP, will facilitate the use of 5F-labeled RNAs in structural, ligand binding, and dynamic studies of RNAs using the advantages of F-19-labeling.