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: 7358941
• 负责人: ZHEN HUANG
• 金额: $ 0.84万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7358941
• 关键词: SELENIUM; DERIVATIZATION; NUCLEIC; ACIDS; RAY

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NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。X射线晶体学是高分辨率测定RNA结构、RNA-蛋白质和DNA-蛋白质复合物结构的有力工具。然而，用重原子衍生化进行相确定是X射线晶体学中长期存在的问题，它在很大程度上减缓了具有新型折叠的核酸的结构确定。促进结构测定的一种方法是用共价连接的重原子标记核酸，这使得能够进行相和结构测定。我们开发了一种新的策略，通过用硒取代氧来衍生RNA和DNA，并通过使用MAD定相方法的X射线晶体学证实了这一原理。我们正在通过化学和酶促合成，衍生DNA和具有新折叠的催化和结合RNA序列，用于核酸及其蛋白质复合物的X射线晶体学研究。