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: 7924777
• 负责人: Grazyna Nowak
• 金额: $ 31.37万
• 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/7924777
• 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：1)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)急性肾功能衰竭后的肾功能修复所涉及的信号事件和调节机制。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目标，因为它们依赖于线粒体功能和氧气能量（ATP）生成，并且具有很大的外源生物转化能力。蛋白激酶信号是调节线粒体功能的主要机制。本提案的目的是确定两种蛋白激酶C同工酶(PKC-?？PKC-?)调节线粒体功能障碍、氧化应激和缺氧后RPTC损伤和修复过程中的细胞存活。我们最近的研究表明，PKC的两种同工异构体在损伤的RPTC修复过程中调节线粒体功能中发挥了关键作用。我们的初步数据证明了PKC-？, PKC - ? ?存在于RPTC线粒体中，PKC磷酸化了一些尚未确定的线粒体蛋白。我们证明了PKC-?？激活介导损伤后线粒体功能障碍。抑制PKC-?？激活促进线粒体功能恢复，减少能量不足，减少缺氧和氧化性损伤后的RPTC坏死。相比之下，PKC-?？减少缺氧和氧化性损伤后的线粒体功能障碍和RPTC坏死。这个提议的中心假设是PKC-?？和PKC - ? ?通过磷酸化线粒体氧化磷酸化装置和/或线粒体通透性过渡孔的关键蛋白来差异调节ATP合成。以下具体目标将检验这一假设。特异性Aim 1将确定PKC-??调控的特定线粒体能量代谢途径。和PKC - ? ?在RPTC损伤和修复期间。特异性Aim 2将识别PKC-?？PKC- ?？调节RPTC损伤线粒体能量代谢。具体目标3将决定PKC-?？和/或PKC - ? ?调控缺血肾线粒体呼吸和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。在这个项目完成后，我们将获得重要的新信息，这将有助于我们理解PKC-?？和PKC - ? ?在损伤后的肾脏修复中，并将为使用这些激酶作为治疗肾功能衰竭的新治疗干预的目标提供见解。公共卫生相关性：目前治疗肾脏和其他器官缺血（氧气供应减少）引起的损伤的疗法是有限的，因为调节肾脏损伤和恢复的机制尚不清楚。本项目将研究三种不同的酶（蛋白激酶）如何调节损伤肾脏的能量产生和肾脏从缺血损伤中恢复。因此，在完成这个项目后，我们将有一个重要的新信息，这将有助于我们理解这些激酶在肾修复中的意义，并将为使用这些激酶作为治疗肾功能衰竭的新疗法的可能性提供见解。