Decoding Hydrogen Peroxide Signaling at Cellular Membranes

解码细胞膜上的过氧化氢信号

• 批准号: 10274736
• 负责人: Jay Thiagarajah
• 金额: $ 44.25万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 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/10274736
• 关键词: Address; Area; Biological; Cell Membrane Permeability; Cell membrane; Cell physiology; Cells; Cellular Membrane; Cysteine; Development; Disease; Environment; Enzymes; Epithelial Cells; Eukaryotic Cell; Future; Generations; Homeostasis; Human; Hydrogen Peroxide; Immune response; Immune signaling; Inflammatory; Malignant Neoplasms; Membrane; Modification; NADPH Oxidase; Nerve Degeneration; Oxidation-Reduction; Pathway interactions; Permeability; Plants; Prevention strategy; Proteins; Proteomics; Reactive Oxygen Species; Regulation; Research; Resolution; Signal Transduction; Signaling Molecule; Stress; Sulfhydryl Compounds; Surface; Tissues; cell motility; environmental change; fluorescence imaging; human disease; migration; novel; protein structure function; tool; trafficking; water channel

PROJECT SUMMARY Eukaryotic cells employ different mechanisms to sense and respond to their environment and maintain tissue homeostasis. Cells have evolved strategies to co-opt stable reactive oxygen species such as hydrogen peroxide (H2O2) as non-transcriptional signaling molecules in order to rapidly respond and adapt to environmental changes. Numerous examples of the influence of H2O2 signaling have emerged, ranging from abiotic stress in plants to immune responses in humans. H2O2 signaling and subsequent regulation of target proteins is therefore an important but still underappreciated biological control mechanism. H2O2-regulated cell signaling is largely dependent on the presence of redox-sensitive thiol switches in protein cysteine residues, where the reactivity of these switches is highly dependent on the local H2O2 concentration. Our previous studies in epithelial cells have shown that key determinants of cellular H2O2 concentrations include generation of H2O2, by cell membrane surface enzymes such as the NADPH oxidases and permeability across cellular membranes which can be facilitated by Aquaporin (AQP) channels. A number of cellular processes such as innate immune signaling, vesicular trafficking and migration have been shown to be regulated by H2O2, but how cellular membranes allow for specific and privileged signaling by H2O2 remains incompletely understood. We therefore propose studies that aim to establish general rules and emergent concepts related to H2O2 signals at membranes. Our studies will encompass four major areas of inquiry that seek to address i) How does plasma membrane permeability to H2O2 influence redox signaling and regulation? ii.) How do H2O2 signals and subsequent regulation of proteins alter essential vesicular trafficking pathways in the cell? iii) How does H2O2 signaling occur at vesicular membranes? iv.) How does spatial control of cellular H2O2 regulate the directional migration of cells? To address these questions, we will apply and develop high resolution quantitative fluorescence imaging to follow the spatial and temporal dynamics of H2O2 signals at membranes, in combination with proteomic approaches to identify target modified cysteines. Further studies will investigate how oxidative modifications alter target protein structure, function and localization, constructing a mechanistic understanding of how H2O2 signals are relayed from cellular membranes. Future studies will build on this framework to uncover strategies to direct and manipulate H2O2 signals for treatment of human disease. Integrated to these studies will be the development of novel tools and approaches to study H2O2 signals at membranes that can be broadly applied for research in this field.

项目概要 真核细胞采用不同的机制来感知和响应其环境并维持组织 体内平衡。细胞已经进化出利用稳定的活性氧（例如氢）的策略 过氧化物（H2O2）作为非转录信号分子，以便快速响应和适应 环境变化。 H2O2 信号传导影响的许多例子已经出现，从 植物的非生物胁迫对人类免疫反应的影响。 H2O2 信号传导和随后的靶标调节 因此，蛋白质是一种重要但仍未得到充分重视的生物控制机制。 H2O2调节细胞 信号传导很大程度上取决于蛋白质半胱氨酸残基中氧化还原敏感硫醇开关的存在， 其中这些开关的反应性高度依赖于局部 H2O2 浓度。我们之前的 对上皮细胞的研究表明，细胞 H2O2 浓度的关键决定因素包括生成 H2O2，通过细胞膜表面酶（例如 NADPH 氧化酶）和跨细胞渗透性 水通道蛋白 (AQP) 通道可以促进膜的形成。许多细胞过程，例如 先天免疫信号、囊泡运输和迁移已被证明受 H2O2 调节，但是 细胞膜如何允许 H2O2 发出特定且优先的信号传导仍不完全清楚。 因此，我们提出旨在建立与 H2O2 相关的一般规则和新兴概念的研究 膜上的信号。我们的研究将涵盖四个主要的探究领域，旨在解决 i) 如何 质膜对 H2O2 的通透性是否会影响氧化还原信号传导和调节？ ii.) H2O2 是如何产生的 信号和随后的蛋白质调节会改变细胞中重要的囊泡运输途径吗？ iii) 如何 H2O2 信号传导发生在囊泡膜上吗？ iv.) 细胞 H2O2 的空间控制如何调节 细胞定向迁移？为了解决这些问题，我们将应用和开发高分辨率 定量荧光成像跟踪膜上 H2O2 信号的空间和时间动态， 结合蛋白质组学方法来鉴定目标修饰的半胱氨酸。进一步的研究将调查 氧化修饰如何改变靶蛋白结构、功能和定位，构建机制 了解 H2O2 信号如何从细胞膜传递。未来的研究将以此为基础 框架来揭示指导和操纵 H2O2 信号以治疗人类疾病的策略。 与这些研究相结合的是开发新的工具和方法来研究 H2O2 信号 膜可广泛应用于该领域的研究。