Mitochondrial Imaging, PTM Analysis and Metabolism Core

线粒体成像、PTM 分析和代谢核心

基本信息

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
项目类别：
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

项目摘要

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。