The role of mitochondrial uncoupling proteins in regulation of reactive oxygen species

线粒体解偶联蛋白在活性氧调节中的作用

• 批准号: 36575480
• 负责人: Professorin Dr. Elena E. Pohl
• 金额: --
• 依托单位: Institut für Physiologie, Pathophysiologie und Biophysik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/36575480?language=en
• 关键词: role; mitochondrial; uncoupling; proteins; regulation

Reactive oxygen species (ROS) produced during mitochondrial respiration are a major cause of the cellular oxidative damage that may underlie neurodegenerative diseases and ageing. Their production is very sensitive to the mitochondrial proton-motive force and may be decreased by uncoupling. In turn, ROS are hypothesized to affect UCP, increasing (i) UCP proton transport rate and (ii) protein abundance. To prove both hypotheses, we measure (i) proton currents through the purified protein reconstituted in planar bilayer membranes and (ii) mRNA and UCP levels in neuronal cells, using RT PCR and confocal microscopy. UCP protection against oxidative damage is evaluated by estimation of ROS amount and mitochondrial potential using fluorescent dyes in cells with different expression levels of protein. To reveal a functional link between uncoupling and protection against oxidative damage, UCPs with various putative functions (UCP1, UCP2, UCP4) are investigated. Differentiation between ROS-mediated UCP activation and upregulation as well as insight into ROS regulation by UCP are expected to provide the basis for a rational drug design, and have, thus, important clinical implications.

在线粒体呼吸过程中产生的活性氧（ROS）是细胞氧化损伤的主要原因，可能是神经退行性疾病和衰老的基础。它们的生产是非常敏感的线粒体质子动力，并可能会减少解偶联。反过来，假设ROS影响UCP，增加（i）UCP质子转运速率和（ii）蛋白质丰度。为了证明这两个假设，我们测量（i）质子电流通过纯化的蛋白质重建在平面双层膜和（ii）mRNA和UCP水平在神经元细胞中，使用RT PCR和共聚焦显微镜。通过使用荧光染料在具有不同蛋白质表达水平的细胞中估计ROS量和线粒体电位来评估UCP对氧化损伤的保护。为了揭示解偶联和保护免受氧化损伤之间的功能联系，研究了具有各种推定功能的UCPs（UCP1，UCP2，UCP4）。ROS介导的UCP激活和上调之间的差异以及UCP对ROS调节的洞察有望为合理的药物设计提供基础，因此具有重要的临床意义。