Quantification of compartmentalized metabolism in eukaryotic systems

真核系统中区室代谢的定量

• 批准号: 10624928
• 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/10624928
• 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; Critical Pathways; Cytoplasm; Cytosol; Data; Diabetes Mellitus; Disease; 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; Ligase; 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; Post-Translational Protein Processing; Process; Proliferating; Publications; Pyruvate; Regulation; Research; Resolution; Route; Signal Transduction; Signaling Molecule; Source; Stable Isotope Labeling; System; Techniques; Testing; Tissues; Toxicology; beta-Hydroxybutyrate; experimental study; inhibitor; lipid biosynthesis; oxidation; preference; thioester; transmission process

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。项目摘要摘要 酰基辅酶A硫代酯（酰基酸）是进化保守的关键代谢中间体， 在真核细胞的不同部分制造并使用。在不同的亚细胞隔室中，酰基-COAS可以 用于满足能量需求，充当信号分子，并用作翻译后的酰基单位 蛋白质的修饰（PTM）。在生物分析化学领域仍未解决的主要挑战 是对这些关键代谢产物的亚细胞局部池的定量，尤其是在 线粒体，核和细胞质中相同的代谢产物可以具有不同的功能。我们将克服 这两个目标的挑战，利用稳定的同位素标记策略，定量质谱法， 药理学，毒理学和独特的细胞生物学模型。首先，我们将量化主要的动力学和命运 细胞和心脏组织中分室化酰基烟的碳底物。其次，我们将量化效果 用组蛋白脱乙酰基酶抑制剂，电子传输链的扰动室特定过程 酰基-COA，下游代谢产物和隔室特异性PTM的抑制剂和遗传模型。这 项目将允许对新陈代谢的不同隔室进行严格的量化，从而定量 在新陈代谢出现问题的正常和病理过程中的研究和操纵。

项目成果

Cumulus cell acetyl-CoA metabolism from acetate is associated with maternal age but only partially with oocyte maturity.

• DOI: 10.1080/19396368.2021.2003479
• 发表时间: 2022-03
• 期刊: Systems biology in reproductive medicine
• 影响因子: 2.4
• 作者: Anderson S;Xu P;Frey AJ;Goodspeed JR;Doan MT;Orris JJ;Clements N;Glassner MJ;Snyder NW
• 通讯作者: Snyder NW

Direct anabolic metabolism of three-carbon propionate to a six-carbon metabolite occurs in vivo across tissues and species.

• DOI: 10.1016/j.jlr.2022.100224
• 发表时间: 2022-06
• 期刊: JOURNAL OF LIPID RESEARCH
• 影响因子: 6.5
• 作者: Doan, Mary T.;Neinast, Michael D.;Varner, Erika L.;Bedi, Kenneth C., Jr.;Bartee, David;Jiang, Helen;Trefely, Sophie;Xu, Peining;Singh, Jay P.;Jang, Cholsoon;Rame, J. Eduardo;Brady, Donita C.;Meier, Jordan L.;Marguiles, Kenneth B.;Arany, Zoltan;Snyder, Nathaniel W.
• 通讯作者: Snyder, Nathaniel W.