Mitochondrial Imaging, PTM Analysis and Metabolism Core

线粒体成像、PTM 分析和代谢核心

• 批准号: 10468113
• 负责人: Stephen M Black
• 金额: $ 31.69万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-20 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10468113
• 关键词: Animals; Apoptotic; Arizona; Biochemical; Bioenergetics; Biological; Biological Assay; Biophysics; Black race; Blood flow; Carnitine; Cells; Cellular Metabolic Process; Child; Complex Analysis; Core Facility; Coupled; Cultured Cells; Data; Detection; Endothelium; Ensure; Equipment; Glycolysis; Goals; Human Resources; Image; In Vitro; Individual; Kinetics; Laboratories; Lung; Mass Spectrum Analysis; Measurement; Mechanical Stress; Membrane Potentials; Metabolic; Metabolism; Mitochondria; Modification; Molecular; Oxygen Consumption; Phenotype; Physiological; Plasma; Post-Translational Protein Processing; Preparation; Procedures; Process; Production; Protein Analysis; Proteins; Pyruvate; Reactive Oxygen Species; Reproducibility; Research Personnel; Resolution; Resources; Running; Sampling; Services; Site; Standardization; Structure; Structure of parenchyma of lung; System; Techniques; Tissues; Training; Universities; Work; congenital heart disorder; cost efficient; experience; fatty acid oxidation; high resolution imaging; in vivo; mechanotransduction; metabolomics; mitochondrial membrane; pulmonary vascular disorder; success; tandem mass spectrometry

ABSTRACT The goal of the Mitochondrial Imaging, PTM analysis, and Metabolism Core (Core B) is to provide a high- quality state-of-the art service to support analyses in all three of the PPG Projects. In this context, Core B will be defined by providing four services. 1) High resolution imaging of mitochondria. This will include analyses of specific protein redistribution to mitochondria, mitochondrial network remodeling, and markers of mitochondrial function (mitochondrial derived reactive oxygen species, mitochondrial membrane potential, carnitine metabolites, ATP levels). 2) Bioenergetics: oxygen consumption, rate of glycolysis, fatty acid oxidation, measurement of cellular ATP kinetics, ETC complex analyses in mechanical stress challenged cells. 3) The analysis of PTMs in biological samples. Individual proteins will be purified to assay PTMs in vivo. To analyze PTMs, we will utilize a mass spectrometry approach coupled to the detection and quantification of specific sites of modification by tandem mass spectrometry. 4) Metabolomic analyses of individual metabolites in cultured cells, lung tissue, and plasma from animals and children born with congenital heart disease that results in changes in pulmonary blood flow (PBF). Core B will enhance the scientific work for all three projects by assuring that all mitochondrial, PTM, and metabolomic analyses are standardized and performed in a uniform standardized manner, and by highly experienced and trained personnel. Core B is run by experienced investigators who have had many years of experience using these techniques. They have developed or adapted many of these techniques and applied them to perform physiological studies in in vitro and in vivo systems. As a central dedicated analytical resource Core B will ensure these analyses are carried out in a cost-efficient manner.

抽象的 线粒体成像、PTM 分析和代谢核心（核心 B）的目标是提供高 高质量、最先进的服务支持所有三个 PPG 项目的分析。在此背景下，Core B 将 通过提供四种服务来定义。 1）线粒体的高分辨率成像。这将包括 分析线粒体的特定蛋白质重新分配、线粒体网络重塑和标记 线粒体功能（线粒体衍生的活性氧、线粒体膜 潜力、肉碱代谢物、ATP 水平）。 2) 生物能学：耗氧量、糖酵解率、脂肪 酸氧化、细胞 ATP 动力学测量、机械应力中的 ETC 复合物分析 受到挑战的细胞。 3）生物样品中PTM的分析。单个蛋白质将被纯化以进行分析 体内 PTM。为了分析 PTM，我们将利用质谱方法结合检测和 通过串联质谱法对特定修饰位点进行量化。 4) 代谢组学分析 动物和出生的儿童的培养细胞、肺组织和血浆中的个体代谢物 导致肺血流量（PBF）变化的先天性心脏病。核心B将增强 通过确保所有线粒体、PTM 和代谢组学分析，为所有三个项目开展科学工作 是标准化的，并以统一的标准化方式执行，并由经验丰富的和 训练有素的人员。 Core B由拥有多年经验的资深调查员负责管理 使用这些技术。他们开发或采用了许多这些技术并将其应用于 在体外和体内系统中进行生理研究。作为中央专用分析资源核心 B 将确保以具有成本效益的方式进行这些分析。