Editing Acetylation and Protein Homeostasis

编辑乙酰化和蛋白质稳态

• 批准号: 10581878
• 负责人: George Burslem
• 金额: $ 25万
• 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/10581878
• 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; 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.

摘要 蛋白质稳态对于维持健康细胞至关重要，并且主要由泛素控制 蛋白酶体系统（UPS），其中蛋白质通过3种酶的级联用泛素标记，导致 被蛋白酶体识别并随后降解。虽然有些蛋白质被组成型识别， 并被该系统降解，其他的则通过翻译后修饰被标记为UPS的底物 例如磷酸化。近年来，非组蛋白的乙酰化作用成为一种重要的机制 泛素-蛋白酶体系统的调节，特别是在E3连接酶底物识别水平。 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 乙酰化驱动蛋白质调节的分子机制和生物学途径 体内平衡（项目1）。此外，基于我们以前针对嵌合体的蛋白水解工作，我们将 开发异双功能的方法来调节蛋白质乙酰化状态作为一种新的机制，以控制 蛋白质稳态的研究，这一基本的生物调节和作为一个潜在的治疗 方法（项目2）。 在项目1中，我们将鉴定和表征在翻译后水平上具有稳定性调节的蛋白质， 乙酰化使用与RNA-Seq配对的蛋白质组学实验，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，我们将表征相关E3对乙酰降解决定子底物的分子识别， 利用生物物理，生物化学和结构方法，揭示了对这种机制的独特见解 蛋白质稳态的关键在项目2中，我们将开发异双功能化合物， 乙酰基转移酶或脱乙酰基酶与新底物结合。基于化学诱导后处理的概念， 翻译修饰，例如蛋白水解靶向嵌合体，我们将确定（去）乙酰化 在设计和合成之前，通过化学生物学方法， 这些项目共同提供了对基础生物学的深入了解， 调节蛋白质稳定性的过程和一种新的化学生物学方法来修饰它们。