Protein Kinase C in the Repair of Cellular Functions

蛋白激酶 C 在细胞功能修复中的作用

• 批准号: 7730427
• 负责人: Grazyna Nowak
• 金额: $ 32.02万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7730427
• 关键词: ATP Synthesis Pathway; Acute Kidney Failure; Adenine Nucleotide Translocase; Blood Circulation; Brain; Brain Hypoxia-Ischemia; Cell Survival; Cell physiology; Cells; Citric Acid Cycle; Complex; Data; Dependence; Electrophoresis; Energy Metabolism; Energy Metabolism Pathway; Enzymes; Event; Gene Delivery; Generations; Goals; Heart; Hypoxia; Immunoprecipitation; In Vitro; Injury; Ischemia; Isoenzymes; Kidney; Kidney Failure; Lipid Peroxidation; Mass Spectrum Analysis; Measures; Mediating; Metabolic Biotransformation; Metabolic Pathway; Mitochondria; Mitochondrial Proteins; Modeling; Morphology; Mus; Myocardial Infarction; Necrosis; Operative Surgical Procedures; Organ; Organ Transplantation; Oxidants; Oxidative Phosphorylation; Oxidative Stress; Oxygen; Pathway interactions; Phosphorylation; Phosphotransferases; Production; Protein Isoforms; Protein Kinase; Protein Kinase C; Proteins; Proteomics; Reactive Oxygen Species; Recovery; Recovery of Function; Regulation; Renal function; Reperfusion Injury; Reperfusion Therapy; Research; Respiration; Role; Signal Transduction; Stroke; Techniques; Testing; Therapeutic Intervention; Tubular formation; Xenobiotics; cell injury; cell type; combat; enzyme activity; improved; in vivo; injured; injury and repair; insight; kidney cell; kidney cortex; mitochondrial dysfunction; mitochondrial permeability transition pore; novel; novel therapeutic intervention; prevent; protein complex; protein function; public health relevance; renal ischemia; repaired; research study; respiratory; restoration; two-dimensional

DESCRIPTION (provided by applicant): The long-term goal of our research is to elucidate the signaling events and regulatory mechanisms involved in: 1) the loss of renal functions associated with acute renal failure (ARF) as caused by ischemia/reperfusion and 2) the repair of renal functions following ARF. The kidney is one of the major target organs for ischemia/reperfusion and oxidative stress-induced damage. Renal proximal tubular cells (RPTC) are a major target of these insults within the kidney due to their dependence on mitochondrial function and oxygen for energy (ATP) generation and large capacity for biotransformation of xenobiotics. Protein kinase signaling is emerging as a major mechanism regulating mitochondrial function. The goal of this proposal is to determine signaling mechanisms through which two protein kinase C isozymes (PKC-??and PKC-?) regulate mitochondrial dysfunction, oxidative stress, and cell survival during injury and repair of RPTC following hypoxia. Our recent studies demonstrated a key role of two isoforms of PKC in regulating mitochondrial functions during repair of injured RPTC. Our preliminary data demonstrate the novel observation that PKC-?, PKC-??are present in RPTC mitochondria and that PKC phosphorylates a number of yet unidentified mitochondrial proteins. We demonstrated that PKC-??activation mediates mitochondrial dysfunction following injury. Inhibition of PKC-??activation promotes recovery of mitochondrial function, diminishes energy deficits, and decreases RPTC necrosis following hypoxia and oxidant-induced injury. In contrast, the activation of PKC-??reduces mitochondrial dysfunction and RPTC necrosis following hypoxia and oxidant-induced injury. The central hypothesis of this proposal is that PKC-??and PKC-??differentially regulate ATP synthesis by phosphorylating key proteins of mitochondrial oxidative phosphorylation apparatus and/or the mitochondrial permeability transition pore. The following specific aims will test this hypothesis. Specific Aim 1 will determine the specific mitochondrial pathways of energy metabolism that are regulated by PKC-??and PKC-??during RPTC injury and repair. Specific Aim 2 will identify proteins through which PKC-??and PKC- ??regulate mitochondrial energy metabolism in injured RPTC. Specific Aim 3 will determine whether PKC-??and/or PKC-??regulate mitochondrial respiration and ATP production in ischemic kidney and whether protein complexes involved in oxidative phosphorylation are mitochondrial targets for PKC-??and/or PKC-??in the kidney in vivo. Upon completion of this project, we will have important novel information that will help us understand the significance of PKC-??and PKC-??in kidney repair following injury and will provide insights into using these kinases as targets for new therapeutic interventions to treat renal failure. PUBLIC HEALTH RELEVANCE: Current therapies to treat injury caused by ischemia (reduced availability of oxygen) in the kidney and other organs are limited because the mechanisms that regulate renal injury and recovery are not well understood. This project will examine how three different enzymes (protein kinases) regulate production of energy in the injured kidney and the recovery of the kidney from injury caused by ischemia. Therefore, upon completion of this project we will have an important novel information that will help us understand the significance of these kinases in renal repair and will provide insights into the possibility of using these kinases as targets for new therapies against renal failure.

描述（由申请人提供）：我们研究的长期目标是阐明涉及以下方面的信号事件和调节机制：1）与缺血/再灌注引起的急性肾功能衰竭（ARF）相关的肾功能丧失和2）ARF后肾功能的修复。肾脏是缺血/再灌注和氧化应激诱导损伤的主要靶器官之一。肾近端小管细胞（RPTC）是肾脏内这些损伤的主要靶点，因为它们依赖于线粒体功能和用于能量（ATP）产生的氧以及用于生物转化外源性物质的大容量。蛋白激酶信号转导正在成为调节线粒体功能的主要机制。这项建议的目标是确定两个蛋白激酶C同工酶（PKC-？？和PKC-？）在缺氧后RPTC损伤和修复期间调节线粒体功能障碍、氧化应激和细胞存活。我们最近的研究表明，在受损的RPTC修复过程中，PKC的两种亚型在调节线粒体功能中起着关键作用。我们的初步数据证实了新的观察结果，PKC-？？存在于RPTC线粒体中，并且PKC磷酸化许多尚未鉴定的线粒体蛋白。我们证明了PKC-？？活化介导损伤后的线粒体功能障碍。抑制PKC-？？活化促进线粒体功能的恢复，减少能量不足，并减少缺氧和氧化剂诱导的损伤后的RPTC坏死。与此相反，PKC-？？减少缺氧和氧化剂诱导的损伤后线粒体功能障碍和RPTC坏死。这个建议的中心假设是，PKC-？？和PKC-？？通过磷酸化线粒体氧化磷酸化装置和/或线粒体通透性转换孔的关键蛋白质来差异调节ATP合成。以下具体目标将检验这一假设。具体目标1将确定具体的线粒体途径的能量代谢的PKC-？？和PKC-？？在RPTC损伤和修复期间。具体目标2将确定蛋白质，通过PKC-？？和PKC-？？调节受损RPTC的线粒体能量代谢。具体目标3将决定是否PKC-？？和/或PKC-？？在缺血性肾脏中调节线粒体呼吸和ATP的产生，以及参与氧化磷酸化的蛋白复合物是否是PKC-？和/或PKC-？？在体内的肾脏中。完成这个项目后，我们将有重要的新信息，这将有助于我们了解PKC的意义-？？和PKC-？？在损伤后的肾修复中的作用，并将为使用这些激酶作为治疗肾衰竭的新治疗干预的靶点提供见解。公共卫生相关性：目前治疗肾和其他器官中由缺血（氧的可用性降低）引起的损伤的疗法是有限的，因为调节肾损伤和恢复的机制还没有很好地理解。该项目将研究三种不同的酶（蛋白激酶）如何调节受损肾脏的能量产生以及缺血引起的肾脏损伤的恢复。因此，在完成本项目后，我们将获得重要的新信息，这将有助于我们了解这些激酶在肾修复中的重要性，并将为使用这些激酶作为肾衰竭新疗法的靶点的可能性提供见解。