Mechanisms of Nucleic Acid Oxidation and Cross-linking

核酸氧化和交联的机制

• 批准号: 0137716
• 负责人: Cynthia Burrows
• 金额: $ 46.3万
• 依托单位: University of Utah
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0137716&HistoricalAwards=false
• 关键词: Mechanisms; Nucleic; Acid; Oxidation; Cross

The objective of this work is to understand the chemistry involved in the formation of DNA-protein cross links. One electron oxidation of DNA fragments generates radical cations at a number of nucleosides which are then trapped by protein nucleophiles. The protein-DNA adducts will then be characterized by a number of spectroscopic techniques including NMR and LC-ESI-MS/MS. Lastly, a 40 mer DNA sequence, and its complement, will be synthesized. This sequence contains a triple G track, a double G and several AT tracks which compliment GC pairs. This strand will then be prepared using G precursors which have been doped with 10% OG (oxidized G) leading to a random distribution of OG in all 10 G positions in the strand and an average of 1 OG/strand. Reactions of these strands with amine and protein nucleophiles and characterization of the subsequent adducts will be used to identify "hot spot" sequences which are most susceptible to adduct formation following oxidative damage. With this award, the Organic and Macromolecular Chemistry Program is supporting the research of Dr. Cynthia J. Burrows of the Department of Chemistry at the University of Utah. Dr. Burrows will explore the reactions of proteins with DNA fragments which have been damaged by oxidative chemical reactions. Uncovering the chemical reactions involved in oxidative DNA damage is important to a fundamental molecular understanding of the chemical processes involved in aging, cancer and a number of neurological disorders. Students trained as a result of working on this project will gain experience in mechanistic organic chemistry as well as biochemistry, hence they will have skills needed by both the biotechnology and pharmaceutical industries.

这项工作的目的是了解 DNA-蛋白质交联形成所涉及的化学过程。 DNA 片段的一个电子氧化在许多核苷上产生自由基阳离子，然后被蛋白质亲核试剂捕获。 然后，蛋白质-DNA 加合物将通过多种光谱技术（包括 NMR 和 LC-ESI-MS/MS）进行表征。 最后，将合成 40 聚体 DNA 序列及其互补序列。 该序列包含一个三重 G 轨道、一个双 G 和几个与 GC 对互补的 AT 轨道。 然后使用掺杂有 10% OG（氧化 G）的 G 前体制备该链，从而导致 OG 随机分布在链的所有 10 个 G 位置上，平均每链 1 个 OG。 这些链与胺和蛋白质亲核试剂的反应以及随后加合物的表征将用于识别氧化损伤后最容易形成加合物的“热点”序列。有机和高分子化学项目通过该奖项支持犹他大学化学系 Cynthia J. Burrows 博士的研究。 Burrows 博士将探索蛋白质与因氧化化学反应而受损的 DNA 片段的反应。 揭示 DNA 氧化损伤中涉及的化学反应对于从分子角度理解与衰老、癌症和许多神经系统疾病相关的化学过程非常重要。 通过该项目接受培训的学生将获得机械有机化学和生物化学方面的经验，因此他们将具备生物技术和制药行业所需的技能。