Probing transcriptional activation at the molecular level - Equipment Supplement

在分子水平上探测转录激活 - 设备补充

• 批准号: 10604581
• 负责人: ANNA K. MAPP
• 金额: $ 10.05万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604581
• 关键词: Affect; Architecture; Binding; Biology; Budgets; Cells; Chemicals; Complex; Data; Equipment; Event; Funding; Future; Genetic Transcription; Goals; Grant; In Vitro; Individual; Intervention; Knowledge; Length; Malignant Neoplasms; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; National Institute of General Medical Sciences; Nature; Pathway interactions; Peptides; Price; Resolution; Running; Structure; Transcription Coactivator; Transcriptional Activation; Work; drug discovery; human disease; inhibitor; instrument; molecular recognition; new therapeutic target; painful neuropathy; peptidomimetics; prediction algorithm; protein protein interaction; small molecule; targeted treatment; therapeutic development; tool

Project Summary The protein‐protein interaction network that form between transcriptional coactivators and activators is dysregulated in every human disease as either a cause or an effect and as such represents a potentially powerful intervention point for therapeutic development. Yet there is little knowledge about this PPI network, particularly how binding events in one domain of a coactivator impacts the global structure and function of the remainder of the domains. We demonstrated in earlier work funded by NIGMS that we can take advantage of the conformational plasticity of coactivators to discover covalent chemical co‐chaperones and small molecules that capture distinct conformational suites of coactivators in vitro and in the cellular milieu. Thus we are poised to answer fundamental questions regarding coactivators: how does a local binding event at an ABD (known to change dynamics and local structure) affect the PPI network of the coactivator, global structural dynamics, and ultimately function. This will not only increase the resolution of our understanding of the basic biology of transcription but also provide a framework by which predictions for the best PPIs for transcription‐targeted therapeutics can be made. Building on the tools and strategies developed in our GM‐funded work, we will define the molecular recognition rules for coactivator ABDs. In doing so, we will produce a suite of selective modulators for functionally distinct coactivators. These data will also provide a rigorous framework and predictive algorithm for the future discovery of selective small molecules and peptidomimetic inhibitors. Through structural studies of intact coactivator‐activator complexes, we will expand the molecular recognition model to include the relationship between local binding‐induced conformational changes and global architectural and functional alterations within full‐length activators and coactivators. We request a replacement for our current automated peptide synthesizer (purchased in 2016) in order to accomplish the goals of the grant, particularly for Project 2. The model we are requesting also has the capability of synthesizing up to four peptides/peptidomimetics sequentially, leading to enormous efficiencies since we can then use the instrument in overnight runs. This will significantly facilitate the accomplishment of the project and due to the unanticipated nature of the need and the price of such an instrument, it cannot be accommodated in the budget of GM136356.

项目摘要 蛋白质相互作用网络，形成转录辅激活因子和 在每一种人类疾病中，激活因子都是失调的原因或结果， 代表了治疗开发的潜在的强有力的干预点。然而有 关于这个PPI网络的知识很少，特别是如何在一个域中绑定事件。 共激活因子影响其余结构域的整体结构和功能。我们 在NIGMS资助的早期工作中证明，我们可以利用 共激活剂的构象可塑性，以发现共价化学共分子伴侣和小分子 在体外和细胞内捕获不同构象的共激活剂的分子 环境。因此，我们准备回答关于共活化剂的基本问题： ABD的局部结合事件（已知会改变动力学和局部结构）影响PPI 网络的共激活剂，全球结构动力学，并最终功能。这不仅会 提高我们对转录的基础生物学的理解的分辨率， 一个框架，通过该框架可以预测转录靶向治疗的最佳PPI， 进行了在全球机制资助的工作中开发的工具和战略的基础上，我们将确定 辅活化剂ABD的分子识别规则。在此过程中，我们将制作一套选择性的 功能上不同的辅活化剂的调节剂。这些数据还将提供一个严格的 框架和预测算法的未来发现的选择性小分子和 拟肽抑制剂。通过对完整的辅活化剂-活化剂复合物的结构研究， 我们将扩展分子识别模型，以包括局部 结合诱导的构象变化和整体结构和功能改变 在全长激活子和辅激活子中 我们要求更换我们目前的自动肽合成仪（2016年购买）， 为了实现赠款的目标，特别是项目2。我们要求的型号 还具有顺序合成多达四种肽/肽模拟物的能力， 因为我们可以在夜间使用仪器。这将 这将极大地促进项目的完成，由于项目的不可预见性， 这种工具的需要和价格，它不能容纳在预算中， GM 136356。