UDE COBRE: SMALL MOLECULE A-HELIX MIMICS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 项目一：宗德洛 螺旋是中央生物识别元件，与蛋白质、DNA和RNA相互作用。用小分子可预测地模拟这种二级结构元素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，但很少有小分子模拟螺旋的例子。这项工作的主要主题是发展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短识别螺旋的模仿上，然后进行扩展的S螺旋的模仿。研究表明，P53、P65和VP16蛋白的酸性激活结构域通过一个短的识别螺旋与它们的目标蛋白相互作用，其中主要的相互作用是通过I、I3和I4位的疏水残基(称为FXXFF基序)进行的。我们将制备基于十氢化氢的支架，它允许呈现螺旋的I、I 1、I 3和I 4残基(一个面)。支架将用来自激活结构域序列的适当侧链来精制。这些分子将被测试与目标蛋白质的结合，以评估支架的适宜性和有效性以及相互作用的特异性。与激活结构域衍生的多肽的竞争实验将确定使用相同的结合位点。模拟物将附着在DNA结合域上，以确定转录激活的有效性。此外，这些分子将被连接到已知的小分子DNA结合基序上，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.