UDE COBRE: SMALL MOLECULE A-HELIX MIMICS

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 项目一：Zondlo 螺旋是与蛋白质、DNA和RNA相互作用的中心生物识别元件。用小分子可预测地模拟这种二级结构元件的能力将允许控制无数的生物事件。 然而，尽管螺旋识别无处不在，但很少有螺旋的小分子模拟物的例子存在。 这项工作的主要主题是开发小分子模拟螺旋及其在生物系统中的应用。 在这个过程中，我们将研究目标识别螺旋的生物学机制。 最初的工作将集中在模仿短识别螺旋，继续模仿扩展的S-螺旋。p53、p65和VP 16蛋白的酸性激活结构域已显示通过短识别螺旋与其靶蛋白相互作用，其中主要相互作用通过位置i、i+3和i+4处的疏水残基（称为FXXFF基序）发生。我们将制备基于十氢化萘的支架，其允许呈现螺旋的i、i+1、i+3和i+4残基（一面）。支架将用来自活化结构域序列的适当侧链进行精心制作。将测试这些分子与靶蛋白的结合，以评估支架的适用性和有效性以及相互作用特异性。与活化结构域衍生肽的竞争实验将确定使用相同的结合位点。模拟物将连接到DNA结合结构域，以确定转录激活的有效性。此外，分子将连接到已知的小分子DNA结合基序，以产生细胞可扩散的小分子转录调节因子。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Project I: Zondlo Helices are central biological recognition elements, interacting with proteins, DNA and RNA. The ability to mimic this secondary structure element predictably with a small molecule would allow control of myriad biological events. However, despite the ubiquity of helical recognition, few examples exist of small molecule mimics of helices. The primary theme of this work is the development of small molecule mimics of helices and their application to biological systems. In the process, we will examine the biological mechanisms of target recognition helics. Initial work will focus on mimicry of short recognition helices, proceeding to mimicry of extended s-helices. The acidic activation domains of the p53, p65 and VP16 proteins have been shown to interact with their target proteins through a short recognition helix, in which the primary interactions occur via hydrophobic residues at positions i, i+3 and i+4 (termed the FXXFF motif). We will prepare decalin-based scaffolds which allow presentation of the i, i+1, i+3 and i+4 residues (one face) of an helix. The scaffolds will be elaborated with appropriate side chains from the activation domain sequences. These molecules will be tested for binding to target proteins to assess scaffold suitability and effectiveness and for interaction specificity. Competition experiments with activation domain-derived peptides will determine that the same binding site is utilized. Mimics will be attached to DNA-binding domains to determine effectiveness in transcription activation. Additionally, the molecules will be attached to known small molecule DNA-binding motifs to generate cell-diffusible small molecule regulators of transcription.