Molecular composition of the mitochondrial permeability transition pore

线粒体通透性转换孔（mPTP）是位于线粒体膜上的一个大的非选择性通道。 线粒体内膜已经确定，在应激期间延长mPTP的开放 条件导致线粒体膜渗透性增加，能量中断 以ATP的形式生成，并最终导致细胞死亡。mPTP开幕是中央事件领先 中风时的组织损伤因此，通过药理学试剂阻断mPTP可以是高度有效的。 保护性的然而，目前对通道（“孔”）部分的分子组成的了解还不够。 mPTP尚未完成。本提案的中心目标是检验以下假设： mPTP的功能性传导通道的形成需要存在非- 无机多磷酸盐和聚羟基丁酸盐的蛋白质聚合物与C- 亚基和Ca ~（2+）。实验将涉及使用几种方法。首先，我们将使用 电生理学以研究纯化的mPTP通道的活性。接下来，我们将调查 mPTP的分子组成和组装，通过使用一些分析方法，包括 免疫化学和质谱。最后，我们将使用野生型和转基因 培养的神经元和稳定的细胞系，研究C-亚基，多磷酸盐和 在活细胞中mPTP活化期间的聚羟基丁酸酯。我们希望我们的研究结果将导致 到详细了解一个最基本和关键的分子过程中， 细胞死亡在未来，这些新知识将为新的治疗策略提供机会， 其将特异性地靶向孔的这些非蛋白质成分， mPTP开放和保护免受组织损伤。

Mitochondrial Permeability Transition Pore (mPTP) is a large non-selective channel located in the mitochondrial inner membrane. It has been established that prolonged opening of mPTP during stress conditions leads to the increase in permeability of the mitochondrial membrane, disruption of energy generation in the form of ATP and eventually to cell death. mPTP opening is the central event leading to tissue damage during stroke. Thus, block of mPTP by pharmacological agents can be highly protective. However, current knowledge of the molecular composition of the channel (“pore”) part of mPTP remains incomplete. The central goal of the current proposal is to test the hypothesis that formation of the functional conducting channel of the mPTP requires the presence of non- proteinaceous polymers of inorganic polyphosphate and polyhydroxybutyrate in combination with the C- subunit and Ca2+. Experiments will involve use of several approaches. First, we will use electrophysiology to study the activity of the purified mPTP channel. Next, we will investigate the molecular composition and assembly of mPTP by using a number of analytical approaches including immunochemistry and mass spectroscopy. Finally, we will use wild-type and genetically modified cultured neurons and stable cell lines to investigate interactions between C-subunit, polyphosphate and polyhydroxybutyrate during mPTP activation in living cells. We expect that results of our study will lead to the detailed understanding of one of the most fundamental and critical molecular processes during cell death. In the future, this new knowledge will open opportunities for novel treatment strategies, which will specifically target these non-proteinaceous components of the pore, effectively prevent mPTP opening and protect against tissue damage.