Quantification of compartmentalized metabolism in eukaryotic systems

真核系统中区室代谢的定量

• 批准号: 9980952
• 负责人: Nathaniel W. Snyder
• 金额: $ 34.08万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9980952
• 关键词: ATP Citrate (pro-S)-Lyase; Acetates; Acetyl Coenzyme A; Acyl Coenzyme A; Address; Amino Acids; Analytical Biochemistry; Biomass; Carbon; Cardiac Myocytes; Cell Fractionation; Cell Nucleus; Cells; Cellular biology; Characteristics; Chemicals; Chemistry; Cholesterol; Citric Acid Cycle; Coenzyme A; Coenzyme A Ligases; Critical Pathways; Cytoplasm; Cytosol; Data; Diabetes Mellitus; Disease; Donor person; Electron Transport; Equilibrium; Eukaryotic Cell; Family member; Fatty Acids; Foundations; Generations; Genetic Models; Glucose; Glutamine; Heart; Heart Transplantation; Heart failure; Histone Acetylation; Histone Deacetylase Inhibitor; Human; Inborn Errors of Metabolism; Isotopes; Ketone Bodies; Kinetics; Knock-out; Label; Letters; Lipids; Liquid Chromatography; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolic; Metabolic Pathway; Metabolism; Methods; Mitochondria; Modeling; Nuclear; Nutrient; Obesity; Output; Oxidation-Reduction; Pantothenic Acid; Pathologic Processes; Pharmacology and Toxicology; Post-Translational Protein Processing; Process; Publications; Pyruvate; Regulation; Research; Resolution; Route; Signal Transduction; Signaling Molecule; Source; Stable Isotope Labeling; System; Techniques; Testing; Tissues; beta-Hydroxybutyrate; experimental study; inhibitor/antagonist; lipid biosynthesis; oxidation; preference; thioester

2. PROJECT SUMMARY ABSTRACT Acyl-coenzyme A thioesters (acyl-CoAs) are evolutionarily conserved critical metabolic intermediates that are made and used in distinct parts of the eukaryotic cell. In different sub-cellular compartments, acyl-CoAs can be used to meet energy needs, act as signaling molecules, and serve as acyl-donors for post-translational modifications (PTMs) of proteins. A major challenge remaining unaddressed in the field of bioanalytical chemistry is the quantitation of the subcellular localized pools of these critical metabolites, especially between the mitochondria, nucleus, and cytosol where the same metabolite can have distinct functions. We will overcome this challenge in two aims, leveraging stable isotope labeling strategies, quantitative mass spectrometry, pharmacology, toxicology, and unique cell biology models. First, we will quantify the kinetics and fate of major carbon substrates for compartmentalized acyl-CoAs in cells and heart tissue. Second, we will quantify the effect of perturbing compartment specific processes with histone deacetylase inhibitors, electron transport chain inhibitors, and genetic models on acyl-CoAs, downstream metabolites, and compartment specific PTMs. This project will allow the rigorous quantification of distinct compartments of metabolism, and thus, their quantitative study and manipulation in normal and pathological processes where metabolism goes awry.

2. 项目摘要