SELENIUM DERIVATIZATION OF NUCLEIC ACIDS FOR X-RAY CRYSTALLOGRAPHY VIA MAD

通过 MAD 对核酸进行硒衍生化，用于 X 射线晶体学

• 批准号: 7726253
• 负责人: ZHEN HUANG
• 金额: $ 2.41万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-18 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7726253
• 关键词: Catalytic DNA; Chemicals; Complex; Computer Retrieval of Information on Scientific Projects Database; Crystallography; DNA; Funding; Grant; Institution; Label; Link; Methods; Nucleic Acids; Oxygen; Phase; Proteins; RNA; RNA Binding; RNA Sequences; Research; Research Personnel; Resolution; Resources; Roentgen Rays; Selenium; Source; Structure; United States National Institutes of Health; X-Ray Crystallography; novel; novel strategies; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. X-ray crystallography is a powerful tool for structure determination of RNA structure, RNA-protein and DNA-protein complexes with high resolution. Derivatization with heavy atoms for phase determination, a long-standing problem in X-ray crystallography, however, has largely slowed down structural determination of nucleic acids with novel folds. One approach to facilitate the structure determination is to label nucleic acids with covalently linked heavy atoms, which enable phase and structure determination. We have developed a novel strategy to derivatize RNA and DNA by replacing oxygen with selenium, and this principle has been demonstrated by X-ray crystallography using MAD phasing method. We are derivatizing DNA and catalytic and binding RNA sequences with novel folds, via chemical and enzymatic synthesis, for X-ray crystallographic studies of nucleic acids and their protein complexes.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 中心，不一定是研究者的机构。 X 射线晶体学是高分辨率 RNA 结构、RNA-蛋白质和 DNA-蛋白质复合物结构测定的强大工具。然而，用于相测定的重原子衍生化是X射线晶体学中长期存在的问题，这在很大程度上减慢了具有新折叠的核酸的结构测定。促进结构确定的一种方法是用共价连接的重原子标记核酸，这使得能够确定相和结构。我们开发了一种新的策略，通过用硒代替氧来衍生 RNA 和 DNA，并且这一原理已通过使用 MAD 定相方法的 X 射线晶体学得到了证明。我们通过化学和酶合成，衍生化具有新颖折叠的 DNA 以及催化和结合 RNA 序列，用于核酸及其蛋白质复合物的 X 射线晶体学研究。