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 在单个MS分析中并行处理多个样本（即样品多路复用）已成为一个 基于MS的系统生物学方法用于翻译医学的首选方法谢谢 具有许多关键优势。首先，单次质谱分析中样品的平行处理 减少任何一个样品所需的材料量。其次，多路复用减少样品制备 和仪器分析时间要求。第三，高度混音的分析最喜欢的生物学系列 复制数据。第四，将样品组合成单个分析的能力允许更少的MS实验，这 反过来允许在所有条件下更大的数据重叠。在此TR＆D中，我们将开发和应用新课程 用于蛋白质和蛋白质后翻译后修饰（PTMS）定量的同种异性标签。具体来说，标签 将建立在胺反应的N，N-二烷基化氨基酸的支架上，该支架与电子兼容 转移解离（ETD）。此目标的最终目标是确定如何最有效地结合 多路复用定量​​的效率具有ETD的优势对不稳定的PTM识别。第二，我们 将开发并应用化学​​标签，以与数据无关采集（DIA）进行多重定量。到 日期，仅代谢标签在DIA的多重分析中才成功。在这里我们将继续 开发我们的基于质量缺陷的化学标签，胺反应性二甲基嘧啶基鸟类（二皮酸）标签， 促进多路复用定量​​直径应用。这些标签最多可提供10 plex dia量化，带来 这种新兴技术的多重能力。第三，我们将改进同质标签的MS硬件 技术。特别是，我们将使用气相离子/离子反应净化目标分析物并去除 污染物，从而提高了同醇标记的定量准确性。而且，为了提高精度，我们将 利用红外照片激活来完全释放所有记者标签，从而增强信号。