INHIBITION OF PROTEIN IMPORT INTO MITOCHONDRIA

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 为了更好地理解心脏病的模型，我们开始努力描述线粒体在细胞凋亡过程中的事件序列。线粒体和疾病之间的关系正变得越来越重要，关于线粒体的结构和功能以及我们的工作试图回答的可能的治疗方法仍然存在许多问题。我们正在澄清的要点是(1)通透性转变是发生在细胞色素c释放之前还是之后，以及(2)如Korsmeyer的研究小组(与Carmen Mannella合作)所建议的那样，内膜是否在通透性转变之后进行了“重塑”以促进细胞色素c的释放。我们使用的是由道格·格林的实验室生产的荧光细胞色素c融合蛋白。我们正在探索的另一个关键问题是线粒体肿胀的时间。目前的一种假设是，由于活性氧物种(ROS)对线粒体的损害通常是由电子传递缺陷造成的。目前正在研究通透性转换、细胞色素c释放和细胞凋亡在ROS损伤中的作用。在与珀金斯博士的合作下，我们正在用EM断层扫描技术对受干扰的线粒体进行结构分析。这项研究有望极大地帮助理解ROS损伤、细胞色素c释放和细胞凋亡过程中结构变化的程度。 另一个项目是基于我们最近在圣地亚哥举行的线粒体疾病会议上与珀金斯博士的新合作。。我们目前的假设是，由于活性氧物种(ROS)对线粒体的损害通常是由电子传递缺陷造成的。通透性转变、细胞色素c释放和细胞凋亡在ROS损伤中的作用正在研究中。在肌营养不良症方面，针对线粒体疾病的免疫-舍弗勒-S细胞培养模型系统正在被应用。此外，乔治城大学的Lee-Jun Wong拥有线粒体疾病患者的活检样本，他的工作正在帮助这些研究。我们还没有通过EM断层扫描获得线粒体的结构分析。然而，这种3-D技术被设想为显著地帮助理解在ROS损伤和凋亡过程中发生的结构改变的程度。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We initiated efforts to characterize the sequence of events in mitochondria during apoptosis to better understand models of heart disease. The relationship between mitochondria and disease is becoming increasingly important, and numerous questions remain regarding mitochondrial structure and function and possible treatments that our work attempts to answer. The important points we are clarifying are (1) Whether the permeability transition occurs before or after cytochrome c release and (2) Whether the inner membrane "remodels" following the permeability transition to facilitate cytochrome c release as suggested by Korsmeyer's group (collaborating with Carmen Mannella). We are using cells transfected with fluorescent cytochrome c fusion proteins produced by Doug Green's lab. Another key issue we are exploring is the timing of mitochondrial swelling. A current hypothesis is that damage to mitochondria due to reactive oxygen species (ROS) is often generated by defective electron transport. The role of the permeability transition, cytochrome c release, and apoptosis in ROS damage are currently being investigated. In collaboration with Dr. Perkins, we are conducting a structural analysis of perturbed mitochondria by EM tomography. This study is envisioned to significantly aid in understanding the extent of structural alterations occurring during ROS damage, cytochrome c release, and apoptosis. Another project is based on our new collaboration with Dr. Perkins coming out of the recent meeting on Mitochondrial Diseases in San Diego. . Our current hypothesis is that damage to mitochondria due to reactive oxygen species (ROS) is often generated by defective electron transport. The role of the permeability transition, cytochrome c release, and apoptosis in ROS damage is being investigated. In relation to muscular dystrophy, Immo Scheffler?s cell culture model systems for mitochondrial diseases are being applied. In addition, the work of Lee-Jun Wong from Georgetown University who has biopsy samples from mitochondrial disease patients is aiding these studies. We have yet to acquire a structural analysis of mitochondria by EM tomography. However, this 3-D technique is envisioned to significantly aid an understanding as to the extent of structural alterations occurring during ROS damage and apoptosis.