Novel approaches to study emerging roles of xenobiotic enzymes

项目摘要/摘要 在过去的75年里，许多研究人员发现饮食摄入/新陈代谢与长期营养不良之间存在明显的联系。 术语健康和疾病。饮食限制(DR)，定义为减少卡路里摄入量，而不是营养不良， 仍然是改善多种物种健康和寿命的最有效和最可重复的干预措施。 不幸的是，长期DR既相对未经测试，也很难在人类身上实施，因此领先 研究人员更好地定义DR改善健康的机制，以努力模仿 在没有真正博士的情况下的好处这个项目专注于一系列异生代谢酶， 含有黄素的单加氧酶，或FMO，在DR下游诱导，最近被报道 这是提高线虫C。 优雅女装。有趣的是，以前的报告也显示在哺乳动物系统中诱导FMO同源物在 糖尿病视网膜病变和其他已知可以延长寿命的疾病。不幸的是，这些影响的机制(S) 保守的FMO蛋白对健康和长寿的影响在很大程度上是未知的，因为它们在I期中起主要作用。 异种生物解毒与观察到的对健康和长寿的影响没有明显的联系。这个项目将 通过开发和利用重要的工具来探索FMO蛋白的内源性作用和底物 测量它们的新陈代谢影响和酶的特定活性。关注最新的代谢组学 通过这项技术，该项目将1)开发一种新的线虫食物来源，以更好地测量线虫的新陈代谢 2)开发一种基于代谢组学的技术，使用氧同位素和 鉴定加氧酶的底物，3)鉴定和验证线虫的候选内源底物 FMO蛋白，以及4)测试线虫内源底物是否也是哺乳动物的底物 FMO蛋白。为了确保它们的成功，这些化验将由线虫和 哺乳动物生物学和衰老，与代谢组学、图谱和数据分析方面的专家合作。这个 由此产生的数据将为线虫生物学家测量基因对新陈代谢的影响提供重要工具 以及环境扰动，这可能会影响蠕虫和细菌生物学，对于生物化学家来说 寻找表征酶底物图谱的新方法。此外，结果主要集中在FMO蛋白上 活动将为出版物和奖励申请编制基础数据，以了解和 探索FMO活性的机制(S)及其改善健康和长寿的作用。

Project Summary/Abstract Many researchers over the past 75 years have found a clear link between dietary intake/metabolism and long- term health and disease. Dietary restriction (DR), defined as a decrease in caloric intake without malnutrition, remains the most potent and reproducible intervention to improve health and longevity across multiple species. Unfortunately, long-term DR is both relatively untested and very difficult to implement in humans, leading researchers to better define the mechanisms through which DR improves health in an effort to mimic the benefits in the absence of true DR. This project focuses on a family of xenobiotic metabolizing enzymes, flavin-containing monooxygenases, or FMOs, that are induced downstream of DR and were recently reported to be both necessary and sufficient to increase health, stress resistance, and longevity in the nematode C. elegans. Interestingly, previous reports also show induction of FMO homologs in mammalian systems under DR and other conditions known to increase longevity. Unfortunately, the mechanism(s) for the effects of these well-conserved FMO proteins on health and longevity are largely unknown, as their primary role in phase I xenobiotic detoxification is not clearly linked to the observed effects on health and longevity. This project will explore the endogenous role and substrates of FMO proteins by developing and utilizing important tools for measuring their metabolic effects and enzyme-specific activities. Focusing on the latest metabolomics technology, the project will 1) develop a novel food source for nematodes to better measure their metabolism on and off xenobiotic compounds, 2) develop a metabolomics based technique to use oxygen isotopes and identify substrates of oxygenases, 3) identify and validate candidate endogenous substrates for nematode FMO proteins, and 4) test whether the nematode endogenous substrates are also substrates of mammalian FMO proteins. To ensure their success, these assays will be performed by experts in nematode and mammalian biology and aging in collaboration with experts in metabolomics profiling and data analysis. The resulting data will provide important tools for nematode biologists measuring the metabolic impact of genetic and environmental perturbations that can effect both worm and bacterial biology and for biochemists looking for new ways to characterize enzymatic substrate profiles. In addition, the results focused on FMO protein activity will produce foundational data for publications and award applications based on understanding and exploiting the mechanism(s) of FMO activity and their role modifying health and longevity.