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: 10417172
• 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/10417172
• 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都是通过蛋白质后翻译修饰在某种程度上介导的。为了 例如，因激酶和磷酸酶催化的异常蛋白磷酸化与A有关 各种各样的癌症。同样，由组蛋白控制的组蛋白蛋白的不管制的乙酰化态 脱乙酰基酶（HDAC）蛋白会导致转录和最终疾病的表观遗传变化。钥匙 表征健康和疾病状态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，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蛋白是药物治疗的靶标。但是，将蛋白质修饰与 下游的生物学活动通常受到限制或不可用，这阻碍了疾病的进展 表征和药物开发。 Pflum实验室中的NIGMS资助的项目解决了开发创新化学的关键需求 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，我们在过去的10年中率先使用ATP类似物对激酶 - 催化的标记反应。在这项先前的工作的基础上，我们在未来5年内提议1） 具有独特能力的新方法，可探测激酶和磷酸酶 - 基底配对以及多种能力 细胞中的蛋白质复合物，以及2）将我们的创新工具应用于各种生物学问题 与多个生物学家合作。在蛋白质乙酰化的工作中，我们在最后一个 两年使用捕获突变体发现HDAC1的非历史基底物的功能， 揭示了HDAC1蛋白在细胞生物学中的意外作用。在接下来的5年中，我们将应用这种强大的 将策略捕获到其他HDAC蛋白同工型中，这将确定HDAC蛋白在 表观遗传学和转录调节以外的活动。鉴于激酶，磷酸酶的关键作用， 和疾病和药物治疗中的HDAC酶，但是可用于研究这些酶 细胞系统中的酶，本应用中提出的化学策略将加强 细胞信号传导和药物设计中的生物医学研究。