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

项目总结 该提案联合了梅奥诊所和威斯康星大学的互补分析专业知识 绘制体力活动(PA)分子图谱的目标。梅奥诊所中心的总体目标是 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 获取MoTrPAC样本的方法。我们提出了一种从非靶向开始的代谢组学工作流程 分析以确定候选代谢物。从那里，注释的代谢物将得到验证，并且是未知的 代谢物将通过新的化合物合成、MS/MS数据和核磁共振相结合来鉴定 光谱学结构解析。从非靶向分析中产生的重要代谢物将指导 随后在血浆和组织样本中进行有针对性的代谢物定量测量。我们提出了一个 蛋白质组学工作流程，包括肌肉、脂肪和外体的全球定量蛋白质组学和 使用磷酸化蛋白质组学、乙酰化蛋白质组学和以下指标评估翻译后修饰 氧化修饰。全球发现蛋白质组学将指导显著的靶向、定量蛋白质组学 血浆中的蛋白质(例如，虹膜蛋白、肌肉生长抑素)。我们的多肽合成能力允许定制合成 用于定量标准的任何稳定的同位素标记的多肽。此外，我们还将测量稳定同位素 血浆、肌肉和单个骨骼肌蛋白质中标记氨基酸的富集性 从而提供了对PA如何影响氨基酸动力学、蛋白质代谢和基因翻译的洞察 转录成单独的蛋白质。血浆和组织中脂质的同位素浓缩将使测量成为可能 脂肪酸动力学对PA的响应。先进的统计方法将融合蛋白质组学和代谢组学 用于生成PA分子概念图的数据。在这里，我们建议利用富有成效的持续 代谢组学和蛋白质组学专家在梅奥和威斯康星大学作为一个单一的 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，同时也贡献我们的肌肉和脂肪组织生物学知识 身体活动联盟的分子传感器(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).