2′-Methylseleno-modified oligoribonucleotides for X-ray crystallography synthesized by the ACE RNA solid-phase approach
2′-Methylseleno-modified oligoribonucleotides for X-ray crystallography synthesized by the ACE RNA solid-phase approach
复制标题
通过 ACE RNA 固相方法合成用于 X 射线晶体学的 2-甲基硒修饰寡核糖核苷酸
作者：
Barbara Puffer;Holger Moroder;M. Aigner;R. Micura
位点特异性修饰的2′-甲基硒基RNA代表了X射线晶体学数据定相的有价值的衍生物。在核酸的三维结构测定中的几个成功应用，例如Diels-Alder核酶，依赖于这种修饰。在这里，我们介绍了所有四个标准核苷，腺苷，胞苷，鸟苷和尿苷的2′-甲基硒代亚磷酰胺构建块的合成路线，这是专为2′-O-双（乙酰氧基乙氧基）甲基（ACE）RNA固相合成。我们还报告了他们纳入寡核糖核苷酸，包括脱保护和纯化。通过ACE RNA化学对2′-甲基硒基标记的方法学扩展是使Se-RNA普遍可获得并广泛传播Se-RNA晶体学研究方法的重要一步。迄今为止，2′-甲基硒修饰的寡核苷酸的制备仅限于2′-O-[（三异丙基硅基）氧基]甲基（TOM）和2′-O-叔丁基二甲基硅基（TBDMS）RNA合成方法。
Site-specifically modified 2′-methylseleno RNA represents a valuable derivative for phasing of X-ray crystallographic data. Several successful applications in three-dimensional structure determination of nucleic acids, such as the Diels–Alder ribozyme, have relied on this modification. Here, we introduce synthetic routes to 2′-methylseleno phosphoramidite building blocks of all four standard nucleosides, adenosine, cytidine, guanosine and uridine, that are tailored for 2′-O-bis(acetoxyethoxy)methyl (ACE) RNA solid-phase synthesis. We additionally report on their incorporation into oligoribonucleotides including deprotection and purification. The methodological expansion of 2′-methylseleno labeling via ACE RNA chemistry is a major step to make Se-RNA generally accessible and to receive broad dissemination of the Se-approach for crystallographic studies on RNA. Thus far, preparation of 2′-methylseleno-modified oligoribonucleotides has been restricted to the 2′-O-[(triisopropylsilyl)oxy]methyl (TOM) and 2′-O-tert-butyldimethylsilyl (TBDMS) RNA synthesis methods.
影响因子：
15
作者：
Du, Q;Carrasco, N;Huang, Z
通讯作者：
Huang, Z
使用 5-溴尿苷取代的 RNA 进行特定的 UV 诱导 RNA 蛋白交联。
影响因子：
2.9
作者：
Gott,JM;Willis,MC;Koch,TH;Uhlenbeck,OC
通讯作者：
Uhlenbeck,OC
将硒共价掺入寡核苷酸中，通过 MAD 测定 X 射线晶体结构：原理证明。
影响因子：
3.9
作者：
Teplova,Marianna;Wilds,ChristopherJ;Wawrzak,Zdzislaw;Tereshko,Valentina;Du,Quan;Carrasco,Nicolas;Huang,Zhen;Egli,Martin
通讯作者：
Egli,Martin