Novel approaches to study emerging roles of xenobiotic enzymes

项目总结/摘要 在过去的75年里，许多研究人员发现饮食摄入/代谢与长期肥胖之间存在明显的联系。 术语健康和疾病。饮食限制（DR），定义为热量摄入减少而不营养不良， 仍然是最有效和可重复的干预措施，以改善多个物种的健康和寿命。 不幸的是，长期DR相对未经测试，并且很难在人类中实施， 研究人员更好地定义DR改善健康的机制，以模仿 在没有真正的DR的情况下的好处。这个项目的重点是一个家庭的异生物质代谢酶， 含黄素的单加氧酶，或FMO，在DR下游诱导，最近报道 是必要的和足够的，以增加健康，抗应激能力，和长寿的线虫C。 优雅有趣的是，先前的报道也显示了在哺乳动物系统中FMO同系物的诱导， DR和其他已知会延长寿命的疾病。不幸的是，这些影响的机制 保守的FMO蛋白对健康和长寿的作用在很大程度上是未知的，因为它们在I期的主要作用是 异生物质解毒与观察到的对健康和寿命的影响没有明确的联系。该项目将 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， 测量它们的代谢效应和酶特异性活性。关注最新的代谢组学 该项目将1）为线虫开发一种新的食物来源，以更好地测量它们的新陈代谢 打开和关闭异生物质化合物，2）开发基于代谢组学的技术以使用氧同位素， 鉴定加氧酶底物，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。为了确保成功，这些试验将由线虫和 哺乳动物生物学和衰老与代谢组学分析和数据分析专家合作。的 由此产生的数据将为线虫生物学家提供重要的工具， 以及环境扰动，这可能会影响蠕虫和细菌的生物学， 寻找表征酶底物谱的新方法。此外，研究结果主要集中在FMO蛋白上， 活动将产生出版物和奖项申请的基础数据， 探索FMO活动的机制及其在改善健康和长寿方面的作用。

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.