Molecular composition of the mitochondrial permeability transition pore

线粒体通透性转换孔(MPTP)是一种位于细胞内的大的非选择性通道。 线粒体内膜。已经证实，在应激过程中MPTP的长时间开放 条件导致线粒体膜通透性增加，能量中断 以三磷酸腺苷的形式产生，最终导致细胞死亡。MPTP开幕式是中心活动的主导 中风时的组织损伤。因此，药理药剂可高度阻断MPTP。 防护性的。然而，目前关于通道(“孔”)部分分子组成的知识 MPTP仍然不完整。当前提案的中心目标是检验以下假设 MPTP功能传导通道的形成需要存在非 无机聚磷酸盐和聚羟基丁酸酯的蛋白质类聚合物与C- 亚基和钙离子。实验将涉及几种方法的使用。首先，我们将使用 电生理学研究纯化的MPTP通道的活性。接下来，我们将调查 MPTP的分子组成和组装使用多种分析方法，包括 免疫化学和质谱学。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以研究C亚单位、多聚磷酸和 活细胞内MPTP激活过程中的聚羟基丁酸。我们期待着我们的研究结果将导致 对过程中最基本和最关键的分子过程之一的详细了解 细胞死亡。在未来，这一新知识将为新的治疗策略提供机会， 它会特别针对毛孔中的这些非蛋白质成分，有效地防止 MPTP打开，防止组织损伤。

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.