DISCOVERY AND DEVELOPMENT OF ANTHRAQUINONE PHOTONUCLEASE

蒽醌光子核酸酶的发现和开发

• 批准号: 2021846
• 负责人: GARY B SCHUSTER
• 金额: $ 10万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-07-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2021846
• 关键词: DNA footprinting; covalent bond; deoxyribonuclease I; laser spectrometry; nuclear magnetic resonance spectroscopy; nucleic acid structure; photochemistry; photolysis; quinones; technology /technique development; ultraviolet radiation

THIS IS A SHANNON AWARD PROVIDING PARTIAL SUPPORT FOR THE RESEARCH PROJECTS THAT FALL SHORT OF THE ASSIGNED INSTITUTE'S FUNDING RANGE BUT ARE IN THE MARGIN OF EXCELLENCE. THE SHANNON AWARD IS INTENDED TO PROVIDE SUPPORT TO TEST THE FEASIBILITY OF THE APPROACH; DEVELOP FURTHER TESTS AND REFINE RESEARCH TECHNIQUES; PERFORM SECONDARY ANALYSIS OF AVAILABLE DATA SETS; OR CONDUCT DISCRETE PROJECTS THAT CAN DEMONSTRATE THE PI'S RESEARCH CAPABILITIES OR LEAD ADDITIONAL WEIGHT TO AN ALREADY MERITORIOUS APPLICATION. THE APPLICATION BELOW IS TAKEN FROM THE ORIGINAL DOCUMENT SUBMITTED BY THE PRINCIPAL INVESTIGATOR. Broadly, the plan is to develop a set of anthraquinone derivatives as photofootprinting agents, focusing on two areas: i) Discovery of quinones which can image major and/or minor groove-bound ligands. ii) Develop time-resolved photofootprinting systems for the study of actively transcribing DNA-RNA polymerase complexes. The reactions of these quinones with nonduplex DNA and RNA forms will be explored to determine if they are able to react selectively with unique DNA and RNA structural features such as loops and hairpins. The quinones will be conjugated with recognition elements, particularly oligonucleotides and peptide nucleic acids, to assess their ability to perform sequence-specific ds cleavage of DNA. And the recently established collaboration with PNA Diagnostics to assess the ability of these quinones to sterilize blood products when they are conjugated with PNAs will be continued. The anthraquinones have special promise as agents for the cleavage of DNA. They can be selectively and rapidly activated by near-UV light. Because of their planar, aromatic skeleton the quinones normally bind to DNA by intercalation between adjacent base pairs. However, subtle structural changes switch them to minor-groove binders and change their reactions with DNA. Excited-state anthraquinone derivatives are good oxidants. Cleavage of DNA is initiated by oxidation of the poly(sugar- phosphate) backbone or by electron transfer from a base. Most importantly, the quinones are stable to repeated cycling through oxidized and reduced forms. Thus the quinones are catalytic and each molecule can cleave numerous DNA strands. The reactions of the quinones will be studied by laser spectroscopy to assess mechanisms for transport of base radical cations (holes) through ds DNA to a site remote and to probe the nature of the initial oxidation step. Analysis of the final cleavage products will be used to reveal the precise site of attack by the quinone. The binding mode of the quinone derivatives will be studied by high resolution NMR spectroscopy.

这是香农奖，为这项研究提供部分支持 项目低于指定研究所的资助范围，但 都在最优秀的边缘。香农奖旨在提供 支持测试该方法的可行性；开发进一步的测试 改进研究方法；对可用的数据进行二次分析 数据集；或执行可演示PI的离散项目 研究能力或导致额外的重量已经有价值的 申请。以下应用程序取自原始文档 由首席调查员提交。 大体上，该计划是开发一套蒽醌衍生品，如 光足迹试剂，重点放在两个领域：i)对苯二酚的发现 其可以成像主要和/或次要沟槽结合的配体。二)发展 用于主动研究的时间分辨照相足迹系统 转录DNA-RNA聚合酶复合体。这些人的反应 将探索具有非双链DNA和RNA形式的喹酮类化合物，以确定 如果他们能够用独特的DNA和RNA结构选择性地做出反应 圈圈和发夹等特征。对苯二酚将与 识别元件，特别是寡核苷酸和肽核 酸，以评估它们执行序列特异性DS切割的能力 关于DNA的。以及最近与PNA诊断公司建立的合作 评估这些对苯二酚在以下情况下对血液制品进行灭菌的能力 它们与PNA的结合将继续下去。 这些蒽醌类化合物具有特殊的前景，可以作为裂解 DNA它们可以被近紫外光选择性地快速激活。 由于其平面的芳香族骨架，对苯二酚通常与之结合。 通过插入相邻碱基对之间的DNA。然而，微妙的 结构变化将它们转换为小凹槽粘结剂，并改变其 与DNA的反应。激发态的蒽醌衍生物是很好的 氧化剂。DNA的切割是由聚(糖-)的氧化启动的。 磷酸盐)主链或通过碱基的电子转移。多数 重要的是，通过氧化，对苯二酚的重复循环是稳定的 和简化形式。因此，对苯二酚是催化的，每个分子都可以 割断大量的DNA链。 将通过激光光谱分析来研究对苯二酚的反应。 评估碱自由基阳离子(空穴)的传输机制 将DNA定向到一个远程位置，并探索初始氧化的性质 一步。对最终裂解产物的分析将用于揭示 对苯二酚攻击的准确位置。对苯二酚的结合方式 将用高分辨率核磁共振波谱对衍生物进行研究。