Neurotoxic Mechanisms of Reactive Nitrogen Intermediates

活性氮中间体的神经毒性机制

基本信息

批准号：
7418655
负责人：
HARRY ISCHIROPOULOS
金额：
$ 27.33万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-04-15 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7418655
关键词：
2-tyrosine 3-nitropropionic acid 3-nitrotyrosine ATP Synthesis Pathway Amino Acids Angiotensin II Animals Apoptosis Apoptotic Architecture Aromatic-L-Amino-Acid Decarboxylases Behavioral Biochemical Biochemistry Biological Models Biology Blood Vessels Bronchoalveolar Lavage Fluid Carboxy-Lyases Cell Line Cell model Cells Cessation of life Chemistry Condition Corpus striatum structure Data Detection Dinitrophenols Disease Disruption Documentation Dopa Dopamine Enzymes Event Family Functional disorder Funding Half-Life Human In Vitro Injection of therapeutic agent Investigation Knowledge Lead Link Lipids Mediator of activation protein Metabolic Pathway Metabolism Methodology Microtubules Mitochondria Mitochondrial Electron Transport Complex II Modeling Molecular Molecular Target Motor Neurons Nature Nerve Degeneration Neurodegenerative Disorders Neuronal Dysfunction Neuronal Injury Neurons Nitrates Nitric Oxide Nitrogen Nitrogen Oxides Oxidants Oxidative Phosphorylation Oxidative Stress Oxidopamine Parkinson Disease Parkinsonian Disorders Pathway interactions Permeability Plasma Positioning Attribute Production Proteins Rattus Reactive Nitrogen Species Regulatory Pathway Reporting Research Personnel Respiration Rodent Role Teratocarcinoma Testing Time Toxic effect Tubulin Tyrosine Tyrosine 3-Monooxygenase Tyrosine Metabolism Pathway Uncoupling Agents Upper arm Work alpha synuclein base beta Tubulin dopaminergic neuron experience hemodynamics in vivo inhibitor/antagonist injured membrane polarity mitochondrial membrane multicatalytic endopeptidase complex neurotoxic neurotoxicity nitrate nitration novel pressure prevent programs protein aggregate protein aggregation protein degradation research study response sound synuclein tyrosyltubulin ligase vascular endothelial dysfunction

项目摘要

DESCRIPTION (provided by applicant): The detection of oxidized proteins, lipids and DNA/RNA in human as well as animal and cellular models of neurodegenerative diseases has been documented. In the last decade additional chemistry based on the formation of reactive nitrogen oxides, byproducts of nitric oxide reactivity have been also documented by the detection of tyrosine-nitrated proteins mostly in the inclusions that characterize Parkinson's and other related diseases. Recent data has also indicated that reactive nitrogen oxides convert the free amino acid tyrosine to 3-nitrotyrosine. This unusual amino acid can be also formed in the CNS following the proteolytic degradation of nitrated proteins. Tyrosine is an important amino acid in the CNS since it serves as the building block for the formation of dopamine. We hypothesize that 3-nitrotyrosine interferes primarily with dopamine formation and tyrosine metabolism. Support for this hypothesis is derived from the sound documentation that 3-nitrotyrosine injected into rodent striatum selectively injures dopaminergic neurons. Another unrecognized feature of 3-nitrotyrosine may be the interference with mitochondrial respiration leading to the decline in ATP synthesis and additional production of reactive species. Preliminary data have also indicated a specific 3- nitrotyrosine-dependent disruption of microtubule assembly leading to the formation of soluble and insoluble protein aggregates. To test the critical aspects of these hypotheses we propose to evaluate the following: 1) determine if 3-nitrotyrosine interferes with the production, metabolism and biology of dopamine and tyrosine, 2) evaluate if 3-nitrotyroinse interferes with mitochondrial respiration and function and 3) examine if the formation of alpha/beta-tubulin aggregates resulting from the specific incorporation of 3-nitroyrosine into a-tubulin serves as a building block for the aggregation and/or fibrilization of alpha-synuclein. An array of biochemical, pharmacological, and molecular approaches that are already in place will be employed to perform the proposed experiments. The proposed experiments will evaluate the critical role of the endogenously generated unusual amino acid, 3-nitrotyrosine, as a central mediator responsible for alterations in fundamental regulatory pathways in dopamine metabolism, oxidative phosphorylation, and protein aggregation, which constitute well-recognized molecular targets responsible for neuronal injury and death in Parkinson's disease and related disorders. Overall a novel and previously unrecognized biological chemistry resulting from the formation of a modified amino acid, 3-nitrotyrosine, will be investigated in order to uncover multifaceted but potentially interrelated pathways that promote neuronal dysfunction in neurodegenerative disorders.

描述（由申请人提供）：人类以及神经退行性疾病的动物和细胞模型中氧化蛋白质、脂质和 DNA/RNA 的检测已被记录。在过去的十年中，基于活性氮氧化物形成的附加化学，一氧化氮反应性的副产物也已通过检测酪氨酸硝化蛋白质（主要存在于帕金森病和其他相关疾病的内含物中）来记录。最近的数据还表明，活性氮氧化物将游离氨基酸酪氨酸转化为 3-硝基酪氨酸。这种不寻常的氨基酸也可以在硝化蛋白质的蛋白水解降解后在中枢神经系统中形成。酪氨酸是中枢神经系统中的重要氨基酸，因为它是形成多巴胺的基本成分。我们假设 3-硝基酪氨酸主要干扰多巴胺形成和酪氨酸代谢。这一假设的支持来自于将 3-硝基酪氨酸注射到啮齿动物纹状体中选择性损伤多巴胺能神经元的可靠文献。 3-硝基酪氨酸的另一个未被认识的特征可能是干扰线粒体呼吸，导致 ATP 合成下降和活性物质的额外产生。初步数据还表明，微管组装的特定3-硝基酪氨酸依赖性破坏导致可溶性和不溶性蛋白质聚集体的形成。为了测试这些假设的关键方面，我们建议评估以下内容：1) 确定 3-硝基酪氨酸是否干扰多巴胺和酪氨酸的产生、代谢和生物学，2) 评估 3-硝基酪氨酸是否干扰线粒体呼吸和功能，3) ）检查 3-硝基酪氨酸特异性掺入 α-微管蛋白所形成的 α/β-微管蛋白聚集体是否可作为构建模块α-突触核蛋白的聚集和/或纤维化。将采用一系列已经到位的生化、药理学和分子方法来进行拟议的实验。拟议的实验将评估内源产生的不寻常氨基酸 3-硝基酪氨酸的关键作用，它作为负责改变多巴胺代谢、氧化磷酸化和蛋白质聚集的基本调节途径的中心介质，这些途径构成了公认的负责多巴胺代谢、氧化磷酸化和蛋白质聚集的分子靶标。用于帕金森病和相关疾病的神经元损伤和死亡。总体而言，我们将研究由修饰氨基酸 3-硝基酪氨酸形成而产生的一种新颖且以前未被认识的生物化学，以揭示促进神经退行性疾病中神经元功能障碍的多方面但可能相互关联的途径。