Mayo Clinic Physical Activity Research Center: Metabolomics and Proteomics Analysis Site

项目摘要 该提案联合了马约诊所和威斯康星州大学的互补分析专业知识， 绘制身体活动（PA）分子图谱的目标。马约临床中心的总体目标是 代谢组学和蛋白质组学（MCCMAPS）是应用最先进的代谢组学和蛋白质组学 接近MoTrPAC样品。我们提出了一个代谢组学工作流程， 分析以鉴定候选代谢物。从那里，注释的代谢物将得到验证，并且未知 代谢物将通过新型化合物合成、MS/MS数据和NMR的组合来鉴定 光谱结构解析。从非靶向分析中出现的重要代谢物将指导 随后在血浆和组织样品中进行靶向定量代谢物测量。我们提出了一个 蛋白质组学工作流程，包括肌肉、脂肪和外泌体中的全局定量蛋白质组学， 使用磷酸蛋白质组学、乙酰蛋白质组学和蛋白质组学指数评估翻译后修饰 氧化修饰全球性的发现蛋白质组学将指导有针对性的，定量的蛋白质组学， 血浆中的蛋白质（例如，鸢尾素、肌肉生长抑制素）。我们的肽合成能力允许定制合成 任何稳定同位素标记的肽作为定量标准。此外，我们将测量稳定同位素 标记氨基酸在血浆、肌肉隔室和个体骨骼肌蛋白中的富集 从而提供了PA如何影响氨基酸动力学，蛋白质代谢和基因翻译的见解 转录到单个蛋白质。血浆和组织中脂质的同位素富集将使测量成为可能 的脂肪酸动力学响应PA。先进的统计方法将融合蛋白质组学和代谢组学 数据以生成PA的分子概念图。在这里，我们建议利用一个富有成效的持续 马约和威斯康星州大学的代谢组学和蛋白质组学专家之间的合作 致力于揭示PA分子特征的综合分析资源。我们的目标不仅是 贡献我们的分析专业知识，也贡献我们的肌肉和脂肪组织生物学知识， 物理活动分子传感器联盟（MoTrPAC）

PROJECT SUMMARY This proposal unites complementary analytical expertise of Mayo Clinic and University of Wisconsin in pursuit of the goal of charting a molecular map of physical activity (PA). The overall goal of the Mayo Clinic Center for Metabolomics and Proteomics (MCCMAPS) is to apply state-of-the-art metabolomics and proteomics approaches to the MoTrPAC samples. We propose a metabolomics workflow that begins with untargeted profiling to identify candidate metabolites. From there, annotated metabolites will be validated, and unknown metabolites will be identified through a combination of novel compound synthesis, MS/MS data and NMR spectroscopy structural elucidation. The important metabolites that emerge from untargeted profiling will guide subsequent targeted quantitative metabolite measurements in plasma and tissue samples. We propose a proteomics workflow that includes global quantitative proteomics in muscle, adipose, and exosomes and assessment of posttranslational modifications using phosphoproteomics, acetylproteomics, and indices of oxidative modifications. Global, discovery proteomics will guide targeted, quantitative proteomics of salient proteins in plasma (e.g., irisin, myostatin). Our peptide synthesis capabilities allows for custom synthesis of any stable isotope labeled peptides for standards for quantitation. In addition, we will measure stable isotope enrichment of labeled amino acids in plasma, muscle compartments, and individual skeletal muscle proteins thus offering an insight on how PA influences amino acid kinetics, protein metabolism, and translation of gene transcripts to individual proteins. Isotopic enrichment of lipids in plasma and tissues will enable measurements of fatty acid kinetics in response to PA. Advanced statistical methods will fuse proteomics and metabolomics data to generate molecular concept maps of PA. Here we propose to leverage a productive ongoing collaboration between metabolomics and proteomics experts at Mayo and University of Wisconsin as a single integrated analytical resource dedicated to uncovering molecular signatures of PA. Our goal is to not only contribute our analytical expertise, but also contribute our knowledge of muscle and adipose tissue biology to the Molecular Transducers of Physical Activity Consortium (MoTrPAC).