CHIRAL PHOSPHATE PROBES OF PROTEIN DNA INTERACTIONS

该项目的主要目标是扩大一个正在进行的 生产性合作，是进一步发展和完善使用 用于蛋白质-DNA高分辨率研究的磷酸盐类似物 交互.这涉及开发新的和改进的方法 用于手性磷酸酯类似物（H- 膦酸酯、硫代磷酸酯和甲烷膦酸酯）和 “校准”这些类似物的效果与一个相对较好的- 特征性蛋白质-DNA复合物（EcoRI内切核酸酶）。的一致性 各种类似物的结果将使我们能够探测蛋白质 与DNA中单个磷酰氧原子的相互作用 解决方案，并测试有关功能角色的详细假设， 这些接触。最后，我们将把这种分析扩展到一个不太好的- 特征性的蛋白质-DNA复合物（BamH I内切核酸酶），以确定 观察到的现象的普遍性。

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.