Editing Acetylation and Protein Homeostasis

编辑乙酰化和蛋白质稳态

• 批准号: 10272903
• 负责人: George Burslem
• 金额: $ 40.63万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10272903
• 关键词: Acetylation; Acetyltransferase; Biochemical; Biological; Biological Process; Biology; Biophysics; Cells; Chemicals; Deacetylase; Disease; EP300 gene; Enzymes; Genetic Transcription; Goals; Health; Human; Molecular; Pathway interactions; Phosphorylation; Post-Translational Protein Processing; Production; Protac; Protein Acetylation; Protein Databases; Proteins; Proteomics; Regulation; Structure; System; Therapeutic; Ubiquitin; Work; design; experimental study; insight; molecular recognition; multicatalytic endopeptidase complex; non-histone protein; novel; novel therapeutic intervention; protein degradation; protein protein interaction; proteostasis; recruit; tool; transcriptome sequencing; ubiquitin-protein ligase

Abstract Protein homeostasis is crucial to maintain healthy cells and is predominantly controlled by the ubiquitin proteasome system (UPS) whereby proteins are tagged with ubiquitin, via a cascade of 3 enzymes, resulting in recognition by the proteasome and subsequent degradation. While some proteins are constitutively recognized and degraded by this system, others are marked as substrates for the UPS by post-translational modifications such as phosphorylation. Recently, acetylation of non-histone proteins has emerged as an important mechanism of regulation for the ubiquitin-proteasome system, particularly at the level of E3 ligase substrate recognition. Leveraging our expertise of the ubiquitin proteasome and protein-protein interactions we propose to elucidate the molecular mechanisms and biological pathways resulting in acetylation driven modulation of protein homeostasis (Project 1). Additionally, building on our previous work with proteolysis targeting chimera, we will develop heterobifunctional approaches to modulate protein acetylation states as a novel mechanism to control protein homeostasis for both the study of this fundamental biological regulation and as a potential therapeutic approach (Project 2). In Project 1, we will identify and characterize proteins with stability regulated at the level of post-translational acetylation. Using proteomics experiments paired with RNA-Seq we will generate a database of proteins with intracellular levels directly controlled by p300 driven acetylation, not altered at the level of transcription. Furthermore, we will characterize the molecular recognition of acetyl degron substrates by the relevant E3 ligases using biophysical, biochemical and structural approaches, revealing unique insights into this mechanism of protein homeostasis. In Project 2, we will develop heterobifunctional compounds which recruit an acetyltransferase or deacetylase to a neo-substrate. Building on the concept of chemically induced post- translational modifications, exemplified by proteolysis targeting chimera, we will identify the (de)acetylation machinery most amenable to this approach via chemical biology approaches before designing and synthesising compounds to edit acetylation in native systems.Together these projects provide insights into basic biological processes regulating protein stability and a novel chemical biology approach to modify them.

抽象的 蛋白质稳态对于维持细胞健康至关重要，并且主要由泛素控制 蛋白酶体系统 (UPS)，通过 3 种酶的级联，蛋白质被泛素标记，从而产生 被蛋白酶体识别并随后降解。虽然某些蛋白质被组成型识别 并被该系统降解，其他则通过翻译后修饰标记为 UPS 的底物 如磷酸化。最近，非组蛋白的乙酰化已成为一种重要机制 泛素-蛋白酶体系统的调控，特别是在 E3 连接酶底物识别水平上。 利用我们在泛素蛋白酶体和蛋白质-蛋白质相互作用方面的专业知识，我们建议阐明 导致乙酰化驱动的蛋白质调节的分子机制和生物途径 体内平衡（项目 1）。此外，在我们之前针对嵌合体的蛋白水解工作的基础上，我们将 开发异双功能方法来调节蛋白质乙酰化状态作为控制的新机制 蛋白质稳态，用于研究这种基本生物调节并作为潜在的治疗方法 方法（项目 2）。 在项目 1 中，我们将鉴定并表征在翻译后水平上受稳定性调节的蛋白质 乙酰化。使用蛋白质组学实验与 RNA-Seq 相结合，我们将生成一个蛋白质数据库 细胞内水平直接由 p300 驱动的乙酰化控制，在转录水平上不改变。 此外，我们将通过相关的 E3 来表征乙酰降解决定子底物的分子识别 使用生物物理、生物化学和结构方法连接酶，揭示了对该机制的独特见解 蛋白质稳态。在项目 2 中，我们将开发异双功能化合物，该化合物招募 乙酰转移酶或脱乙酰酶至新底物。基于化学诱导后的概念 翻译修饰，例如靶向嵌合体的蛋白水解，我们将鉴定（去）乙酰化 在设计和合成之前通过化学生物学方法最适合这种方法的机械 化合物在天然系统中编辑乙酰化。这些项目共同提供了对基本生物学的见解 调节蛋白质稳定性的过程以及修改它们的新颖的化学生物学方法。