Chemical Approaches to Study Protein Post-Translational Modifications

研究蛋白质翻译后修饰的化学方法

• 批准号: 10164804
• 负责人: Mary Kay H Pflum
• 金额: $ 50.62万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10164804
• 关键词: Acetylation; Address; Biological; Biomedical Research; Cells; Cellular biology; Chemicals; Collaborations; Development; Disease; Drug Design; Enzymes; Epigenetic Process; Event; Funding; Genetic Transcription; HDAC1 gene; Histone Deacetylase; Histones; Label; Lead; Link; Malignant Neoplasms; Mediating; Methods; Molecular; Multiprotein Complexes; National Institute of General Medical Sciences; Pharmacotherapy; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Post-Translational Protein Processing; Protein Acetylation; Protein Isoforms; Proteins; Reaction; Role; Signal Transduction; System; Transcriptional Regulation; Work; analog; drug development; epigenetic regulation; innovation; link protein; mutant; protein function; targeted treatment; tool

Summary Most biological events in the cell are mediated at some level by protein post-translational modifications. For example, aberrant protein phosphorylation catalyzed by kinase and phosphatase enzymes is linked to a wide variety of cancers. Similarly, the unregulated acetylation state of histone proteins, controlled by histone deacetylase (HDAC) proteins, can lead to epigenetic changes in transcription and ultimately disease. Key to characterizing both healthy and disease states is a detailed molecular understanding of the role of protein post-translational modifications, such as phosphorylation and acetylation, on protein function and interactions. Importantly, enzymes regulating post-translational modifications, including kinase, phosphatase, and HDAC proteins, are targets of drug treatment. Yet, tools linking protein modifications to downstream biological activities are often limited or unavailable, which has stalled progress in disease characterization and drug development. The NIGMS-funded projects in the Pflum lab address the critical need to develop innovative chemical approaches to discover unanticipated functions of protein modifying enzymes in cell biology. In our work with protein phosphorylation, we have pioneered in the last 10 years use of ATP analogs for kinase- catalyzed labeling reactions. Building on this prior work, we propose in the next 5 years to 1) develop a new suite of methods with unique abilities to probe kinase- and phosphatase-substrate pairs and multi- protein complexes in cells, and 2) apply our innovative tools to a variety of biological problems in collaboration with multiple biologists. In our work with protein acetylation, we have demonstrated in the last two years the power of using trapping mutants to discover non-histone substrates of HDAC1, which has revealed unexpected roles of HDAC1 proteins in cell biology. In the next 5 years, we will apply this powerful trapping strategy to additional HDAC protein isoforms, which will establish the role of HDAC proteins in activities beyond epigenetics and transcriptional regulation. Given the critical role of kinase, phosphatase, and HDAC enzymes in disease and drug treatment, yet the inadequate tools available to study these enzymes in cellular systems, the enabling chemical strategies proposed in this application will strengthen biomedical research in cell signaling and drug design.

概括 细胞中的大多数生物事件在某种程度上是由蛋白质翻译后修饰介导的。为了 例如，由激酶和磷酸酶催化的异常蛋白质磷酸化与 多种癌症。同样，组蛋白不受调控的乙酰化状态，由组蛋白控制 脱乙酰酶 (HDAC) 蛋白可导致转录的表观遗传变化并最终导致疾病。钥匙 表征健康和疾病状态的关键是对蛋白质作用的详细分子理解 翻译后修饰，例如磷酸化和乙酰化，对蛋白质功能和 互动。重要的是，调节翻译后修饰的酶，包括激酶， 磷酸酶和 HDAC 蛋白是药物治疗的目标。然而，将蛋白质修饰与 下游生物活性往往有限或不可用，这阻碍了疾病的进展 表征和药物开发。 Pflum 实验室 NIGMS 资助的项目解决了开发创新化学品的迫切需求 发现细胞生物学中蛋白质修饰酶的意外功能的方法。在我们的工作中 通过蛋白质磷酸化，我们在过去 10 年中率先使用 ATP 类似物进行激酶- 催化标记反应。基于之前的工作，我们建议在未来 5 年内 1) 开发一个 一套新的方法具有独特的能力来探测激酶和磷酸酶底物对和多 细胞中的蛋白质复合物，2) 将我们的创新工具应用于解决各种生物问题 与多名生物学家合作。在我们对蛋白质乙酰化的研究中，我们在过去的研究中证明了 两年后，我们使用捕获突变体来发现 HDAC1 的非组蛋白底物，该底物已 揭示了 HDAC1 蛋白在细胞生物学中意想不到的作用。未来5年，我们将应用这一强大的 对其他 HDAC 蛋白亚型的捕获策略，这将确定 HDAC 蛋白在 表观遗传学和转录调控之外的活动。鉴于激酶、磷酸酶的关键作用， 和 HDAC 酶在疾病和药物治疗中的应用，但可用于研究这些的工具不足 细胞系统中的酶，本申请中提出的有利化学策略将加强 细胞信号传导和药物设计的生物医学研究。