Regulation of O-GlcNAcylation During Injury

损伤期间 O-GlcNAc 酰化的调节

基本信息

批准号：
9751373
负责人：
Natasha Elizabeth Zachara
金额：
$ 40.94万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9751373
关键词：
Acetylation Acute Address Affect American Antibodies Biochemical Cardiac Cardiotonic Agents Cardiovascular Diseases Cell Survival Cell model Cessation of life Cytoplasmic Protein Data Development Disease model Enzymes Etiology Excision Fatty-acid synthase Functional disorder Goals HSPB1 gene Health Heart Heat Stress Disorders Hemorrhage Human Hypertension Hypoxia Infarction Injury Ischemia Ischemic Preconditioning Knowledge Lead Link Malignant Neoplasms Maps Mass Spectrum Analysis Mediating Metabolic syndrome Mitochondrial Proteins Modeling Modification Molecular Monosaccharides Mus Myocardial Infarction Myocardial Ischemia Nerve Degeneration Nuclear Proteins O-GlcNAc transferase Organ Oxidative Stress Pathway interactions Patients Pharmacology Phosphorylation Play Post-Translational Protein Processing Proteins Regulation Reperfusion Injury Reperfusion Therapy Role Site Stroke Substrate Specificity Techniques Tissues Trauma Work Writing base biological adaptation to stress cardioprotection cell injury cytotoxicity endoplasmic reticulum stress improved in vitro Model in vivo inhibitor/antagonist injured insight new therapeutic target novel novel therapeutics peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase protein protein interaction response response to injury sugar

项目摘要

Summary The modification of intracellular proteins by O-linked β-N-acetylglucosamine (O-GlcNAc) has emerged as a novel regulator of cytoprotection1-4. Numerous forms of cellular injury, including cardiac ischemic preconditioning (acute, prolonged, and remote), lead to elevated levels of O-GlcNAc in both in vivo and in vitro models. Elevating O-GlcNAcylation before, or immediately after, the induction of cellular injury is protective in models of ischemia reperfusion (I/R) injury, as well as heat stress, oxidative stress, endoplasmic reticulum stress, hypoxia, and trauma hemorrhage2,5-8. Together, these data suggest that O-GlcNAc is a novel endogenous cardioprotective agent. To date, the majority of work studying the O-GlcNAc modification in models of I/R injury has focused on identifying the proteins and mechanisms by which O-GlcNAc mediates cardioprotection3,4. However, understanding the regulation of the O-GlcNAc modification during injury is critical and is yet unstudied. The goal of this proposal is to map the regulatory networks of the enzymes that cycle the O-GlcNAc modification, the O- GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Specifically, we will complete the following aims: Aim 1. Define the impact of protein-protein interactions on OGT and OGA activity and substrate targeting in the injured heart. Quantitative mass spectrometry will be used to identify protein-interactors of OGT and OGA. A combination of biochemical approaches will be used to interrogate the role of these effector- proteins on O-GlcNAc cycling in the ischemic heart. Aim 2. Map the OGT and OGA covalent-regulatory networks in the injured heart. The O-GlcNAc modification sites, and other covalent-regulators, of OGT and OGA will be identified. The impact of O- GlcNAcylation on OGT/OGA function will be assessed in models of oxidative stress and I/R injury. Collectively, we anticipate that these studies will define the pathways that regulate OGT, OGA, and O- GlcNAcylation during I/R injury. This critical insight will provide a framework for investigating novel therapeutic targets for myocardial infarction and an understanding about how the O-GlcNAc-mediated stress response is dysregulated contributing to cardiovascular disease

概括通过O连接的β-N-乙酰葡萄糖（O-GLCNAC）对细胞内蛋白的修饰已成为A 细胞保护作用的新型调节剂1-4。许多形式的细胞损伤，包括心脏缺血性预处理（急性，延长和远程），导致体内和体外模型中的O-GlCNAC水平升高。抬高在缺血模型中保护细胞损伤之前或之后的O-Glcnacylation。再灌注（I/R）损伤，以及热应激，氧化应激，内质网应激，缺氧和缺氧和创伤出血2,5-8。总之，这些数据表明O-GLCNAC是一种新型的内源性心脏保护代理人。迄今识别O-GLCNAC介导心脏保护的蛋白质和机制3,4。然而，了解受伤期间O-GLCNAC修饰的调节至关重要，并且仍然没有研究。目标该建议的内容是绘制循环O-GLCNAC修饰的酶的调节网络，O- GlcNAC转移酶（OGT）和O-Glcnacase（OGA）。具体来说，我们将完成以下目的： AIM 1。定义蛋白质 - 蛋白质相互作用对OGT和OGA活性和底物靶向的影响在受伤的心中。定量质谱法将用于鉴定OGT的蛋白质相互作用和OGA。生化方法的结合将用于审问这些效应的作用。缺血性心脏中O-GLCNAC循环的蛋白质。 AIM 2。绘制受伤心脏中的OGT和OGA共价调节网络。 O-GLCNAC 将确定OGT和OGA的修改位点以及其他共价调节器。 O-的影响 OGT/OGA功能上的Glcnacylation将在氧化应激和I/R损伤的模型中评估。总的来说，我们预计这些研究将定义调节OGT，OGA和O-的途径。 I/R损伤期间的Glcnacylation。这种关键见解将为研究新疗法提供一个框架心肌梗塞的靶标以及对O-GLCNAC介导的应力反应的理解导致心血管疾病的失调