CHIRAL PHOSPHATE PROBES OF PROTEIN DNA INTERACTIONS

蛋白质 DNA 相互作用的手性磷酸盐探针

• 批准号: 2292059
• 负责人: LINDA JEN-JACOBSON
• 金额: $ 2.52万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-05-01 至 1997-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2292059
• 关键词: DNA; chemical synthesis; endonuclease; intermolecular interaction; phosphates; phosphonate; proteins; stereochemistry; thiophosphate

The primary objective of this project, an expansion of an ongoing productive collaboration, is to further develop and refine the use of phosphate analogues for the high-resolution study of protein-DNA interactions. This involves the development of new and improved methods for the stereospecific synthesis of chiral phosphate analogues (H- phosphonates, phosphorothioates and methanephosphonates) and the "calibration" of the effects of these analogues with a relatively well- characterized protein-DNA complex (Eco RI endonuclease). The concordance of results with the various analogues will permit us to probe protein interactions with individual phosphoryl oxygens in DNA at atomic resolution, and test detailed hypotheses about the functional roles of these contacts. Finally, we will extend this analysis to a less well- characterized protein-DNA complex (Bam HI endonuclease) to determine the generality of the observed phenomena.

该项目的主要目标是扩展正在进行的 富有成效的协作，是进一步开发和完善对 用于蛋白质-DNA 高分辨率研究的磷酸盐类似物 互动。这涉及新方法和改进方法的开发 用于手性磷酸类似物（H- 膦酸盐、硫代磷酸盐和甲膦酸盐）和 用相对良好的“校准”这些类似物的效果 表征了蛋白质-DNA 复合物（Eco RI 核酸内切酶）。索引 各种类似物的结果将使我们能够探测蛋白质 与DNA中单个磷酰氧在原子上的相互作用 分辨率，并测试有关功能作用的详细假设 这些接触。最后，我们将把这个分析扩展到不太好的情况—— 表征蛋白质-DNA 复合物（Bam HI 核酸内切酶）以确定 观察到的现象的普遍性。