Developing Mass Spectrometry-based Approaches to Characterize Mono- and Poly(ADP-ribosyl)ated Proteomes

开发基于质谱的方法来表征单和多（ADP-核糖基）化蛋白质组

• 批准号: 10458521
• 负责人: Isabel Uribe
• 金额: $ 4.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-12-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10458521
• 关键词: ADP ribosylation; Adenosine Diphosphate Ribose; Amino Acids; Biological; Biological Process; Cell physiology; Cells; Chemistry; Complex; DNA; DNA Damage; DNA Repair; Data; Disease; Enzymes; Equilibrium; Excision; Glycoside Hydrolases; Histones; Investigative Techniques; Label; Length; Malignant Neoplasms; Mass Spectrum Analysis; Methods; Modeling; Modification; Molecular Conformation; Nerve Degeneration; Outcome; Pathogenesis; Peptides; Physiological; Physiological Processes; Polymerase; Polymers; Post-Translational Protein Processing; Process; Proteins; Proteome; Proteomics; Publishing; Recombinant Proteins; Regulation; SWI1; Sepharose; Site; Solid; Specificity; Structure; System; aspartylglutamate; base; biological adaptation to stress; cell growth regulation; chemical addition; chemical group; chemical property; cofactor; human disease; inorganic phosphate; insight; monomer; novel; peptide A; phosphodiester; preservation; scaffold; tool

PROJECT SUMMARY Post-translational modifications (PTMs) involve the addition of chemical groups to proteins, modulating the structures or functions of the modified proteins to coordinate cellular processes. ADP-ribosylation, the addition of adenosine diphosphate ribose to proteins, exists in both a monomeric (MARylation) and polymeric (PARylation) form, allowing for more complex regulation. The balance between the synthesis and degradation of ADP-ribosylation forms is fundamental for regulating cellular processes. Disruption of this balance leads to human diseases like cancer and neurodegeneration. However, much remains to be learned about the respective contributions of MARylation and PARylation to the regulation of physiological processes and disease pathogenesis. Mass spectrometry (MS)-based proteomics methods are the go-to techniques for investigating PTMs in healthy and disease states. Previously published proteomics methods studying ADP-ribosylation have been successful in identifying modification sites on amino acids of varied chemical properties. However, all of these methods fail to identify the ADP-ribosylation form associated with each site. This proposal aims to develop a MS-based proteomics method to identify the site and form of ADP-ribosylation on a proteome-wide level. To preserve information about the form of ADP-ribosylation at modification sites, my method will either (a) keep ADP- ribosylation intact during enrichment or (b) determine whether the amino acid is MARylated or PARylated. With a tool to distinguish between the ADP-ribosylation forms at each site, we can identify potential functional differences of MARylated and PARylated proteins in healthy and disease states. Ultimately, this workflow will provide valuable insights into the biological functions of the different forms of ADP-ribosylation in physiological and disease states.

项目摘要 翻译后修饰（PTM）涉及将化学基团添加到蛋白质中， 调节修饰蛋白的结构或功能以协调细胞过程。 ADP-核糖基化，将腺苷二磷酸核糖添加到蛋白 单体（MARYLATION）和聚合物（parylation）形式，可以进行更复杂的调节。 ADP-核糖基化形式的合成和降解之间的平衡是至关重要的 调节细胞过程。这种平衡的破坏会导致癌症等人类疾病 神经变性。但是，关于各自的贡献仍有很多待了解 对生理过程和疾病的调节的玛利亚和抚养 发病。质谱（MS）的蛋白质组学方法是用于首选技术 研究健康和疾病状态的PTM。以前发表的蛋白质组学方法 研究ADP-核糖基在识别氨基酸上的修饰位点已成功 各种化学性质的。但是，所有这些方法都无法识别ADP-核糖化 与每个站点关联的形式。该建议旨在开发一种基于MS的蛋白质组学方法 在蛋白质组水平上识别ADP-核糖基化的位点和形式。保留信息 关于在修饰位点的ADP-核糖基化形式，我的方法将（a）保持ADP- 富集过程中核糖基化完整或（b）确定氨基酸是玛丽化还是 加油。使用一个工具来区分每个站点的ADP-核糖基化形式，我们可以 确定在健康和 疾病状态。最终，该工作流将提供对生物学功能的宝贵见解 生理和疾病状态中ADP-核糖基化的不同形式。