Role of Prohibitin Nitrosylation in its Neuroprotective Functions

抑制素亚硝基化在其神经保护功能中的作用

• 批准号: 10626154
• 负责人: Ping Zhou
• 金额: $ 42.37万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10626154
• 关键词: Address; Affect; Astrocytes; Binding; Bioenergetics; Biological Assay; Brain; Brain Injuries; Brain Pathology; Cells; Cerebral Ischemia; Cerebrum; Coculture Techniques; Code; Crista ampullaris; Cysteine; Electron Microscopy; Embryo; Environment; Event; Gel; Genus Hippocampus; Glucose; In Vitro; Ischemia; Ischemic Brain Injury; Ischemic Neuronal Injury; Knock-in; Knock-in Mouse; Laboratories; Life; Link; Maintenance; Mediating; Messenger RNA; Mitochondria; Mitochondrial Proteins; Modeling; Molecular; Mus; Mutant Strains Mice; Mutate; Mutation; Neuroglia; Neurologic; Neurons; Nitric Oxide; Nitric Oxide Donors; Oxidative Phosphorylation; Oxygen; Pathology; Patients; Pattern; Phosphorylation; Play; Positioning Attribute; Post-Translational Protein Processing; Process; Production; Protein S; Proteins; Reactive Oxygen Species; Regulation; Respiratory Chain; Role; S-Nitrosoglutathione; Scaffolding Protein; Somatic Mutation; Specificity; Stress; Stroke; Structure; System; Testing; Therapeutic; Transgenic Mice; Work; artery occlusion; brain tissue; cell type; deprivation; design; in vivo; inhibitor; ischemic injury; mitochondrial dysfunction; mutant; nervous system disorder; neuron loss; neuronal survival; neuroprotection; novel; prevent; prohibitin; protein protein interaction; restoration; stroke risk; targeted treatment; therapy design; translational potential; treatment strategy

Project Summary/Abstract The mitochondrial protein Prohibitin (PHB) is essential for life. Its importance to cellular activities is attested by the fact that deletion of PHB is embryonic lethal in mice and that to date no mutation has been found in the coding region of PHB in any neurological disease conditions, indicating that PHB integrity is essential and that somatic mutation is detrimental. Our recent work has demonstrated that PHB has remarkable neuroprotective potential against ischemic brain injury with an underlying mitochondrial associated mechanism. Its expression is critical for mitochondrial function in stress situations. However, how this important protein that are stable at both mRNA and protein levels, is functionally regulated in neuroprotection, as well as how it is dysregulated in other neurological conditions, remain surprisingly unknown. In exploring the mechanisms of PHB regulation, we discovered that nitric oxide (NO) is required for PHB expression mediated neuroprotection. Therefore, we investigated the interaction between NO and PHB and found that NO modifies PHB post-translationally, through protein s-nitrosylation, a novel regulatory mechanism similar to protein phosphorylation. In this application, we propose to study the effects of PHB S-nitrosylation and the mechanisms underlying functional regulation of PHB by NO. Our central hypothesis is that nitrosylation is critical for PHB’s neuroprotective function and, consequently, disturbances of PHB nitrosylation are detrimental and contribute to pathology. We will use a novel mutant knock-in mouse, in which the sole cysteine residue of PHB protein is mutated so that PHB cannot be nitrosylated, to analyze the mechanisms of NO regulation and the effects of loss of PHB nitrosylation on PHB function, in the settings of brain ischemic injury in association with mitochondria structural alterations. Three specific aims will systematically test the hypothesis. The results of the proposed studies will reveal a previously unrecognized regulatory mechanism of PHB which we believe is crucial to facilitate the design of potential therapies that could ultimately benefit patients at risk of stroke and other neurological diseases.

项目总结/摘要 线粒体蛋白质Prohibitin（PHB）是生命所必需的。它对细胞活动的重要性 通过以下事实证明，即在小鼠中缺失PHB是胚胎致死的，并且迄今为止， 在任何神经疾病的情况下， 表明PHB完整性是必需的，体细胞突变是有害的。我们最近 研究表明，PHB具有显著的神经保护潜力， 脑损伤与潜在的线粒体相关机制。它的表达对于 线粒体在应激状态下的功能然而，这种重要的蛋白质是如何稳定在 mRNA和蛋白质水平，在神经保护中功能性调节，以及它是如何 在其他神经系统疾病中的失调，仍然令人惊讶地未知。在探索 通过对PHB调控机制的研究，我们发现，一氧化氮（NO）是PHB调控所必需的， 表达介导的神经保护作用。因此，我们研究了NO 发现NO通过蛋白质s-亚硝基化， 类似于蛋白质磷酸化的新的调节机制。在本申请中，我们提出 研究PHB-S-亚硝基化的影响及其功能调控机制 我们的中心假设是，亚硝基化是关键的PHB的 神经保护功能，因此，干扰的PHB亚硝基化， 有害并导致病理学。我们将使用一种新的突变敲入小鼠， 其中突变了PHB蛋白的唯一半胱氨酸残基，使得PHB不能被亚硝基化， 分析NO的调控机制及PHB亚硝酰化缺失对PHB的影响 功能，在脑缺血性损伤的情况下与线粒体结构相关 改变。三个具体目标将系统地检验这一假设。的结果 拟议的研究将揭示一种以前未被认识到的PHB调节机制， 我相信这对促进潜在疗法的设计至关重要， 有中风和其他神经系统疾病风险的患者。