Targeted Synaptic Proteomics

靶向突触蛋白质组学

• 批准号: 8245702
• 负责人: JUNMIN PENG
• 金额: $ 19.07万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8245702
• 关键词: Automatic Data Processing; Biological; Cell model; Collaborations; Communication; Communities; Complex Mixtures; Computer software; Core Protein; Data; Data Set; Detection; Development; Epilepsy; Etiology; Event; Generic Drugs; Goals; Health; Huntington Disease; Isotopes; Label; Laboratories; Learning; Light; Liquid Chromatography; Mass Spectrum Analysis; Measurement; Measures; Membrane; Memory; Mental Depression; Methods; Modification; Molecular; Monitor; Mouse Protein; Mus; Nerve Degeneration; Neurons; Neurosciences; Neurotransmitter Receptor; Peptides; Phosphorylation; Physiological; Play; Post-Translational Protein Processing; Postsynaptic Membrane; Protein Dynamics; Proteins; Proteomics; Reaction; Reagent; Regulation; Reproducibility; Research; Role; Sampling; Series; Serine; Shotguns; Signal Transduction; Site; Stable Isotope Labeling; Substance abuse problem; Synapses; Synaptic plasticity; Technology; Testing; Threonine; Tissues; Tyrosine; Ubiquitination; Universities; Work; base; density; genetic regulatory protein; insight; mouse model; multiple reaction monitoring; nervous system disorder; postsynaptic; postsynaptic density protein; research study; tandem mass spectrometry

DESCRIPTION (provided by applicant): The goal of this research is to develop a quantitative strategy for systematically monitoring the abundance and modifications of postsynaptic proteins. The postsynaptic density (PSD) is a specialized membrane structure essential for neuronal communication. Dynamic protein changes in the PSD play a pivotal role in synapse plasticity. Dysregulation of PSD proteins has been implicated in the etiology of a variety of neurological disorders, such as depression, epilepsy, substance abuse and neurodegeneration. Approximately 1,000 proteins including neurotransmitter receptors and regulatory proteins have been identified in the PSD, but a quantitative, dynamic view of the protein components is still missing. Much less is known about the modulation of posttranslational modifications (e.g. phosphorylation) in the PSD. The main challenge is how to consistently quantify the PSD proteins and their functional modifications under different physiological and pathological conditions. We hypothesize that targeted proteomics will provide a simple, large-scale, and sensitive measurement of proteins and modifications in the PSD. Targeted proteomics uses a specific mass spectrometry technology termed Selected Reaction Monitoring (SRM) or Multiple Reaction Monitoring (MRM). The targeted proteomics overcomes a number of caveats associated with conventional discovery mass spectrometry, such as limited reproducibility, data redundancy and compromised sensitivity. We will first obtain comprehensive PSD datasets and then develop an SRM-based strategy to quantify core PSD proteins, as well as associated phosphorylation events. The strategy will be thoroughly tested to profile PSD components in a cellular model of synaptic activation and during the development of Huntington disease. The study will establish an SRM-based strategy for targeted quantitative PSD analysis. The strategy is expected to greatly simplify current proteomics platform, and will provide crucial basis for understanding molecular mechanisms of learning, memory and neurodegeneration. PUBLIC HEALTH RELEVANCE: Molecular regulation of synapse is the key to the formation of learning and memory. We propose to develop a simplified state-of-the-art mass spectrometry technology to quantitatively monitor the dynamics of proteins and posttranslational modifications in the postsynaptic compartment. The methods will provide a generic approach for investigating the synaptic plasticity under diverse physiological and pathological conditions.

描述（由申请人提供）：本研究的目标是开发一种定量策略，用于系统地监测突触后蛋白的丰度和修饰。突触后密度（PSD）是一种特殊的膜结构，对神经元的通讯至关重要。PSD蛋白的动态变化在突触可塑性中起着关键作用。PSD蛋白的失调与多种神经系统疾病的病因有关，如抑郁症、癫痫、药物滥用和神经变性。在PSD中已经发现了大约1000种蛋白质，包括神经递质受体和调节蛋白，但对蛋白质成分的定量、动态观察仍然缺失。对PSD中翻译后修饰（如磷酸化）的调节知之甚少。主要的挑战是如何在不同的生理和病理条件下一致地量化PSD蛋白及其功能修饰。我们假设靶向蛋白质组学将为PSD中的蛋白质和修饰提供一种简单、大规模和敏感的测量方法。靶向蛋白质组学使用一种特定的质谱技术，称为选择反应监测（SRM）或多反应监测（MRM）。靶向蛋白质组学克服了传统发现质谱法的一些缺点，如重复性有限、数据冗余和灵敏度降低。我们将首先获得全面的PSD数据集，然后开发一个基于srm的策略来量化核心PSD蛋白，以及相关的磷酸化事件。该策略将被彻底测试，以在突触激活的细胞模型和亨廷顿病的发展过程中分析PSD成分。该研究将建立一个基于srm的策略，用于有针对性的定量PSD分析。该策略有望极大地简化现有的蛋白质组学平台，并将为理解学习、记忆和神经变性的分子机制提供重要的基础。