天然样交联双链RNA的合成及其性质研究。
Synthesis of native-like crosslinked duplex RNA and study of its properties.
作者信息
Onizuka Kazumitsu, Hazemi Madoka E, Thomas Justin M, Monteleone Leanna R, Yamada Ken, Imoto Shuhei, Beal Peter A, Nagatsugi Fumi
机构信息
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi 980-8577, Japan.
Department of Chemistry, University of California, Davis, One Shields Ave, Davis, CA 95616, USA.
出版信息
Bioorg Med Chem. 2017 Apr 1;25(7):2191-2199. doi: 10.1016/j.bmc.2017.02.034. Epub 2017 Feb 21.
A variety of enzymes have been found to interact with double-stranded RNA (dsRNA) in order to carry out its functions. We have endeavored to prepare the covalently crosslinked native-like duplex RNA, which could be useful for biochemical studies and RNA nanotechnology. In this study, the interstrand covalently linked duplex RNA was formed by a crosslinking reaction between vinylpurine (VP) and the target cytosine or uracil in RNA. We measured melting temperatures and CD spectra to identify the properties of the VP crosslinked duplex RNA. The crosslinking formation increased the thermodynamic stability without disturbing the natural conformation of dsRNA. In addition, a competitive binding experiment with the duplex RNA binding enzyme, ADAR2, showed the crosslinked dsRNA bound the protein with nearly the same binding affinity as the natural dsRNA, confirming that it has finely preserved the natural traits of duplex RNA.
已发现多种酶与双链RNA（dsRNA）相互作用以执行其功能。我们致力于制备共价交联的天然双链RNA，这可能对生化研究和RNA纳米技术有用。在本研究中，链间共价连接的双链RNA通过乙烯基嘌呤（VP）与RNA中的目标胞嘧啶或尿嘧啶之间的交联反应形成。我们测量了熔解温度和圆二色光谱以确定VP交联双链RNA的性质。交联的形成增加了热力学稳定性，而不会干扰dsRNA的天然构象。此外，与双链RNA结合酶ADAR2的竞争性结合实验表明，交联的dsRNA与蛋白质的结合亲和力与天然dsRNA几乎相同，证实其精细地保留了双链RNA的天然特性。
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