Functional Studies of Endocannabinoid Metabolizing Enzymes

内源性大麻素代谢酶的功能研究

基本信息

批准号：
8484801
负责人：
BENJAMIN F CRAVATT
金额：
$ 48.9万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8484801
关键词：
2-arachidonylglycerol Active Sites Agonist Amides Anti-Inflammatory Agents Anti-inflammatory Behavior Binding Binding Proteins Biochemical Biochemical Pathway Biological Brain Carrier Proteins Catalysis Cell Cycle Kinetics Cells Chemicals Chemistry Degradation Pathway Disease Drug Targeting Drug usage Endocannabinoids Environment Enzymatic Biochemistry Enzymes Ethanolamines Fatty Acids Funding Genetic Goals Human Hydrolase Hydrolysis Individual Kinetics Knowledge Link Lipid A Lipid Bilayers Lipids Maps Mediating Membrane Membrane Lipids Mental Depression Metabolic Metabolic Diseases Metabolism Methods Modeling Molecular Biology Monoacylglycerol Lipases Mus Mutation Nervous system structure Neurons Obesity Pain Pathway interactions Physiological Physiology Play Process Property Proteins Proteomics Recruitment Activity Role Signal Transduction Site Structure Synthesis Chemistry System TRP channel Taurine Testing Travel anandamide base chemical genetics enzyme structure factor A fatty acid amide hydrolase in vivo inhibitor/antagonist insight metabolomics mouse model mutant protein transport tool uptake

项目摘要

TPhreo jeocbt jescutmivme aoryf : Project 1 is to understand the biochemical and cellular mechanisms for terminating endocannabinoid (EC) signaling in the nervous system. The two major ECs, anandamide (AEA) and 2- arachidonoylglycerol (2-AG), are degraded by disfinct enzymatic pathways in vivo. AEA is principally regulated by the integral membrane enzyme fatty acid amide hydrolase (FAAH), which also hydrolyzes several other bioactive fatty acid amides, including the anti-infiammatory lipid A/-palmitoyl ethanolamine, the satiating factor A/-oleoyl ethanolamine, and the TRP channel agonist A/-arachidonoyl taurine. A major challenge in understanding FAAH-regulated fatty acid amide signaling is to discriminate the distinct physiological functions of endogenous FAAH substrates. 2-AG is hydrolyzed by several enzymes in the brain, including the soluble enzyme monoacylglycerol lipase (MAGL) and the integral membrane hydrolases ABHD6 and ABHD12. A major challenge in understanding 2-AG signaling is to determine the unique biochemical and cellular properties of individual 2-AG hydrolytic enzymes. Prior to enzymatic hydrolysis, both AEA and 2-AG may interact with binding proteins responsible for cellular uptake and transport. A major challenge in understanding EC uptake and transport is to identify the putative proteins that mediate these processes. In the previous funding period, we used a combination of synthetic chemistry, enzymology, molecular biology, and functional proteomic/metabolomic methods to enrich our understanding of EC degradative pathways. We have also developed powerful new chemical and genetic tools to probe the function of these pathways. In this renewal application, we plan to use our suite of chemical and genetic tools to gain further mechanistic insights into EC degradative pathways. Specifically, we aim to: 1) characterize mouse models with altered EC degradative pathways, 2) characterize the biochemical and cell biological properties of brain 2-AG hydrolases, and 3) map lipid-protein interactions in EC degradative pathways using chemical proteomic methods. We anticipate that these studies will enhance our understanding of the biochemical pathways that terminate EC signaling in the nervous system and define key components in these pathways that may serve as drug targets for the treatment of a range of human disorders, including pain, depression, and metabolic disorders.

TPhreo jeocbt jescutmivme aoryf ：项目 1 是了解终止的生化和细胞机制神经系统中的内源性大麻素 (EC) 信号传导。两种主要的 EC，anandamide (AEA) 和 2- 花生四烯酰甘油 (2-AG) 在体内通过不同的酶途径降解。 AEA主要受到监管由整合膜酶脂肪酸酰胺水解酶 (FAAH) 产生，该酶还水解其他几种物质生物活性脂肪酸酰胺，包括抗炎脂质 A/-棕榈酰乙醇胺、饱足因子 A/-油酰乙醇胺和TRP通道激动剂A/-花生四烯酰牛磺酸。一个重大挑战了解 FAAH 调节的脂肪酸酰胺信号传导是为了区分不同的生理功能内源性 FAAH 底物。 2-AG 被大脑中的多种酶水解，包括可溶性酶单酰基甘油脂肪酶 (MAGL) 和整合膜水解酶 ABHD6 和 ABHD12。一个了解 2-AG 信号传导的主要挑战是确定独特的生化和细胞特性单个2-AG水解酶。在酶水解之前，AEA 和 2-AG 都可能与负责细胞摄取和运输的结合蛋白。了解 EC 吸收的主要挑战运输的目的是识别介导这些过程的推定蛋白质。在上一个资助期内，我们结合了合成化学、酶学、分子生物学和功能学蛋白质组学/代谢组学方法丰富了我们对 EC 降解途径的理解。我们还有开发了强大的新化学和遗传工具来探测这些途径的功能。在这次更新中应用程序中，我们计划使用我们的化学和遗传工具套件来获得对 EC 的进一步机制见解降解途径。具体来说，我们的目标是：1）表征具有改变的 EC 降解的小鼠模型途径，2) 表征脑 2-AG 水解酶的生化和细胞生物学特性，以及 3) 图谱使用化学蛋白质组学方法研究 EC 降解途径中的脂质-蛋白质相互作用。我们预计这些研究将增强我们对终止 EC 信号传导的生化途径的理解神经系统并定义这些途径中的关键组成部分，这些组成部分可以作为治疗的药物靶标一系列人类疾病，包括疼痛、抑郁和代谢紊乱。