Training in proteomics of novel kinase substrates for neurodegeneration

神经变性新型激酶底物蛋白质组学培训

• 批准号: 7334031
• 负责人: Charleen T Chu
• 金额: $ 21.12万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7334031
• 关键词: Affect; Animal Model; Award; Binding; Biochemistry; Biological; Brain; Cell Death; Cell Survival; Cells; Cessation of life; Collaborations; Complex; Condition; Consultations; Core Facility; Data; Development; Disease; Docking; Dopaminergic Cell; Educational workshop; Engineering; Environment; Family; Free Radicals; Future; Genes; Genetic; Goals; Grant; Hand; Harvest; Human; Injury; Knowledge; Laboratories; Lead; Life; Link; Localized; Maps; Mass Spectrum Analysis; Mentors; Mentorship; Midbrain structure; Mitochondria; Modeling; Mutagenesis; Mutate; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Operative Surgical Procedures; Oxidation-Reduction; Parkinson Disease; Parkinsonian Disorders; Pathogenesis; Pathway Analysis; Pharmacologic Substance; Phosphoproteins; Phosphorylation; Phosphotransferases; Physicians; Pilot Projects; Point Mutation; Principal Investigator; Protein Kinase; Protein Overexpression; Proteins; Proteomics; Publishing; RNA Interference; Radiolabeled; Recombinants; Regulation; Research; Research Personnel; Research Training; Role; Scientist; Signal Pathway; Signal Transduction; Standards of Weights and Measures; Stress; Structural Models; Structure; Techniques; Technology; Testing; Toxin; Training; Translating; Universities; Validation; Work; analog; base; brain tissue; career; data mining; day; design; dopaminergic neuron; experience; extracellular; injured; insight; instrument; interdisciplinary collaboration; mass spectrometer; metabolomics; mouse model; multidisciplinary; mutant; neuron loss; neuronal survival; neuroprotection; novel; radiotracer; tool

DESCRIPTION (provided by applicant): Parkinson disease (PD) is a debilitating neurodegenerative disease for which there are no effective neuroprotective therapies. Converging data from post-mortem brain studies, oxidative toxin models, and PD genetics all implicate altered mitochondrial kinase networks in PD pathogenesis. Mutations in the mitochondrial PTEN-induced kinase 1 (PINK1) occur in familial and possibly sporadic PD. The candidate is an established and productive investigator in the signaling regulation of neuronal fate in culture and animal models of PD pathogenesis. The candidate's published and preliminary work implicate mitochondrial targeting of extracellular signal regulated protein kinase 2 (ERK2) and decreased PINK1 signaling in promoting dopaminergic neuronal cell death. Overexpression of wild type PINK1 confers neuroprotection in cell death models. Thus, identifying downstream targets of PINK1, and of mitochondrial targets of redox-activated ERK2, would represent important steps in understanding mechanisms that regulate neuronal survival and death. Identification of substrates for novel kinases such as PINK1, and of context specific ERK2 targets, can be accomplished using the powerful techniques of phospho-proteomics and mass spectroscopy. This proposal capitalizes on the strong cell signaling, proteomic/metabolomic, pharmaceutical, and neurodegeneration research environments at the University of Pittsburgh. The candidate's immediate goals are to obtain training in mass spectrometry and phospho-proteomics under the mentorship of Billy Day, Director of the Pittsburgh Proteomic Core Facility. Co-mentor Bruce Freeman, expert in free radical biochemistry, will provide complementary training in basic redox proteomics. In addition to short courses, seminars, and technical workshops, practical experience will be derived from a pilot project involving ATP-binding pocket mutagenesis and mass spectrometry to identify substrates of PINK1. Lay summary: This career enhancement award will provide an established physician-scientist studying neurodegenerative diseases with essential cross-training in highly specialized target identification technologies. Solidifying interdisciplinary collaborations will enable the principal investigator to more rapidly translate knowledge on factors that determine whether or not neurons can successfully adapt to disease promoting stresses into novel targets for designing neuroprotective PD therapies.

描述（申请人提供）：帕金森病（PD）是一种使人衰弱的神经退行性疾病，目前尚无有效的神经保护疗法。来自死后脑研究、氧化毒素模型和帕金森病遗传学的数据都暗示了帕金森病发病过程中线粒体激酶网络的改变。线粒体pten诱导的激酶1 （PINK1）突变发生在家族性和可能的散发性PD中。候选人是在PD发病机制的培养和动物模型中神经元命运的信号调节方面建立和富有成效的研究者。该候选人已发表的和初步的工作表明，线粒体靶向细胞外信号调节蛋白激酶2 （ERK2）和减少PINK1信号传导促进多巴胺能神经元细胞死亡。野生型PINK1过表达在细胞死亡模型中具有神经保护作用。因此，鉴定PINK1的下游靶点，以及氧化还原激活的ERK2的线粒体靶点，将是理解调节神经元存活和死亡机制的重要步骤。使用强大的磷酸化蛋白质组学和质谱技术，可以鉴定PINK1等新型激酶的底物和上下文特定的ERK2靶点。该提案利用了匹兹堡大学强大的细胞信号、蛋白质组学/代谢组学、药物和神经变性研究环境。候选人的近期目标是在匹兹堡蛋白质组学核心设施主任Billy Day的指导下获得质谱和磷酸化蛋白质组学的培训。共同导师Bruce Freeman是自由基生物化学方面的专家，他将提供基础氧化还原蛋白质组学方面的补充培训。除了短期课程、研讨会和技术讲习班外，还将从一个涉及atp结合口袋诱变和质谱鉴定PINK1底物的试点项目中获得实际经验。概要：该职业提升奖将为研究神经退行性疾病的资深内科科学家提供高度专业化目标识别技术的必要交叉培训。巩固跨学科合作将使首席研究者能够更快地将决定神经元是否能够成功适应疾病促进压力的因素转化为设计神经保护性PD治疗的新靶点。