CHIRAL PHOSPHATE PROBES OF PROTEIN DNA INTERACTIONS

该项目的主要目标是对正在进行的 富有成效的协作，是为了进一步开发和细化使用 用于蛋白质-DNA高分辨率研究的磷酸盐类似物 互动。这涉及到开发新的和改进的方法。 用于立体定向合成手性磷酸类似物(H- 磷酸盐、硫代磷酸盐和甲基磷酸盐)和 对这些类似物的效果进行相对较好的“校准”- 具有特征的蛋白质-DNA复合体(Eco RI内切酶)。和谐共鸣 各种类似物的结果将使我们能够探测蛋白质 在原子水平上与DNA中各磷酰氧的相互作用 解决，并测试关于功能角色的详细假设 这些联系人。最后，我们将把这个分析扩展到一个不太好的地方- 鉴定蛋白质-DNA复合体(Bam HI内切酶)以确定 观察到的现象的概括性。

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.