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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要线粒体蛋白抑制素 (PHB) 对于生命至关重要，它对细胞活动非常重要。事实证明，删除 PHB 对小鼠胚胎是致命的，而且迄今为止还没有发现在任何神经系统疾病中，PHB 编码区均发现突变，表明 PHB 完整性至关重要，而体细胞突变是有害的。研究表明，PHB 具有显着的抗缺血神经保护潜力脑损伤与其潜在的线粒体机制相关。其表达对于脑损伤至关重要。然而，线粒体在应激情况下的功能如何保持稳定。 mRNA 和蛋白质水平在神经保护中受到功能调节，以及它是如何调节的在其他神经系统疾病中的失调，在探索过程中仍然令人惊讶地未知。 PHB调节机制，我们发现一氧化氮（NO）是PHB所必需的因此，我们研究了 NO 之间的相互作用。和 PHB，发现 NO 通过蛋白质 s-亚硝基化在翻译后修饰 PHB，在本申请中，我们提出了类似于蛋白质磷酸化的新调控机制。研究PHB S-亚硝基化的影响和功能调节的机制我们的中心假设是亚硝基化对于 PHB 至关重要。神经保护功能以及因此导致的 PHB 亚硝基化紊乱我们将使用一种新型突变基因敲入小鼠。其中PHB蛋白的唯一半胱氨酸残基发生突变，使得PHB不能被亚硝基化，分析NO调节机制以及PHB亚硝基化缺失对PHB的影响功能，在与线粒体结构相关的脑缺血损伤的情况下三个具体目标将系统地检验假设的结果。拟议的研究将揭示我们之前未认识到的 PHB 调节机制相信对于促进最终受益的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。