National Center for Quantitative Biology of Complex Systems

国家复杂系统定量生物学中心

• 批准号: 10426384
• 负责人: LINGJUN LI
• 金额: $ 17.28万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-05 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10426384
• 关键词: Alanine; Amines; Amino Acids; Biocompatible Materials; Biological; Biology; Biomedical Technology; Cells; Chemicals; Collection; Complex; Data; Data Analyses; Defect; Dissociation; Electron Transport; Emerging Technologies; Gases; Goals; Ions; Isotopes; Label; Lead; Leucine; Light; Liquid substance; Mass Spectrum Analysis; Measures; Metabolic; Methods; Mission; Modernization; Mus; Noise; Ornithine; Peptides; Phase; Post-Translational Protein Processing; Preparation; Production; Proline; Proteins; Proteome; Reaction; Reporter; Resources; Sampling; Signal Transduction; Stable Isotope Labeling; Structure; System; Systems Biology; Techniques; Technology; Testing; Time; base; chemical reaction; design; experimental study; human tissue; improved; instrument; novel; novel strategies; parallel processing; photoactivation; prevent; scaffold; success; tandem mass spectrometry; translational medicine

PROJECT SUMMARY – TR&D 2 Processing multiple samples in parallel in a single MS analysis – i.e., sample multiplexing – has become a preferred method for the application of MS-based systems biology approaches to translational medicine thanks to numerous key advantages. First, parallel processing of samples in a single mass spectrometric analysis reduces the amount of material required from any one sample. Second, multiplexing reduces sample preparation and instrument analysis time requirements. Third, highly plexed analyses facilitate collection of biological replicate data. Fourth, the ability to combine samples into a single analysis permits fewer MS experiments, which in turn allows for greater data overlap across all conditions. In this TR&D, we will develop and apply a new class of isobaric tags for protein and protein post-translational modifications (PTMs) quantitation. Specifically, the tags will be built upon an amine-reactive N,N-dialkylated amino acid-based scaffold that is compatible with electron transfer dissociation (ETD). The ultimate goal of this aim is to determine how to most effectively combine the efficiencies of multiplexed quantification with the advantages of ETD for labile PTM identification. Second, we will develop and apply chemical tags for multiplexed quantification with data independent acquisition (DIA). To date, only metabolic labels have been successful in multiplexing analysis for DIA. Here we will continue to develop our mass defect-based chemical tag, amine-reactive dimethyl pyrimidinyl ornithine (DiPyrO) tags, to facilitate multiplexed quantitative DIA applications. These tags will offer up to 10-plex DIA quantification, bringing multiplexed capacity for this emergent technology. Third, we will improve MS hardware for isobaric tag technology. In particular, we will use gas-phase ion/ion reactions to purify the target analyte and remove contaminants, thereby improving the quantitative accuracy of isobaric tagging. And, to improve precision, we will leverage infrared photo-activation to completely release all reporter tags, thereby boosting signal.

项目摘要 – TR&D 2 在单次 MS 分析中并行处理多个样品（即样品多重分析）已成为一种 基于 MS 的系统生物学方法应用于转化医学的首选方法，感谢 首先，在单个质谱分析中并行处理样品。 其次，多重分析减少了样品制备 第三，高度复杂的分析有利于生物的收集。 第四，将样品合并到单一分析中的能力允许更少的 MS 实验，从而减少重复数据的数量。 反过来，在这个 TR&D 中，我们将开发并应用一个新的类别。 用于蛋白质和蛋白质翻译后修饰 (PTM) 定量的同量异位标签。 将建立在与电子兼容的胺反应性 N,N-二烷基化氨基酸支架上 该目标的最终目标是确定如何最有效地结合转移解离（ETD）。 其次，我们利用 ETD 进行多重定量的效率来进行不稳定的 PTM 鉴定。 将开发和应用化学标签，通过数据独立采集（DIA）进行多重定量。 迄今为止，只有代谢标签在 DIA 多重分析中取得了成功，在这里我们将继续。 开发基于质量缺陷的化学标签，胺反应性二甲基嘧啶基鸟氨酸 (DiPyrO) 标签， 促进多重定量 DIA 应用，这些标签将提供高达 10 重的 DIA 定量。 第三，我们将改进同量异位标签的 MS 硬件。 特别是，我们将使用气相离子/离子反应来纯化和去除目标分析物。 污染物，从而提高同量异位标记的定量准确性，并且，为了提高精度，我们将。 利用红外光激活完全释放所有报告标签，从而增强信号。