Mitochondrial PKC epsilon in cardiac protection

描述(由申请人提供)：一氧化氮(NO)供体通过涉及蛋白激酶Ce(PKCE)的信号机制保护心肌缺血/再灌注损伤。PKCE针对的下游效应者进行保护的定义仍然不明确。最近的一项研究表明，腺嘌呤核苷酸转运子(ANT1)-线粒体通透性转换(MPT)孔的核心单位--是PKC(.目前的研究重点是确定临床上相关的心脏保护药物是否没有供体通过调节MPT孔的ANT1成分起作用，并检查PKCE直接修饰孔在这一过程中的作用。这项研究将使用标准的线粒体功能分析，以测量MPT的孔活动，标准的生化程序，以确定蛋白质的表达和相互作用，以及最先进的质谱学，以检测ANT1被PKC(。中心假设是，在NO供体诱导的心脏保护中，PKCE的一个关键任务是降低线粒体通透性转换的倾向。该作用部分涉及对MPT孔的核心成分ANT1的直接修饰。其具体目的是：(1)阐明依赖PKCE的线粒体通透性转换调节在NO诱导的小鼠心脏保护模型中的作用。(2)确定MPT孔的重要成分ANT1是否是PKC下游的分子靶点(并鉴定在NO诱导的心肌保护过程中被PKCE磷酸化的ANT1的特定氨基酸残基)。

DESCRIPTION (provided by applicant): Nitric oxide (NO) donors protect against myocardial ischemia/reperfusion injury via signaling mechanisms that involve protein kinase C e (PKCe). The downstream effectors targeted by PKCe to carry out protection remain poorly defined. A recent study has implicated the adenine nucleotide translocator (ANT1)---a core unit of the mitochondrial permeability transition (MPT) pore as a signaling partner of PKC(. The focus of the current investigation is to determine whether NO donors, clinically relevant cardiac protective drugs, act via modulation of the ANT1 component of the MPT pore and to examine the role of direct modification of the pore by PKCe in this process. The study will employ standard mitochondrial function assays, to measure MPT pore activity, standard biochemistry procedures, to determine protein expression and interactions, and state-of-the-art mass spectrometry, to examine phosphorylation of ANT1 by PKC(. The central hypothesis is that a critical task of PKCe in NO donor-induced cardiac protection is to reduce the propensity for mitochondrial permeability transition. This action involves, in part, direct modification of a core component of the MPT pore, ANT1. The specific aims are: (1) To elucidate the role of PKCe-dependent regulation of mitochondrial permeability transition in a murine model of NO-induced cardiac protection. (2) To determine whether ANT1, an essential element of the MPT pore, is a downstream molecular target of PKC( and to identify the specific amino acid residues of ANT1 phosphorylated by PKCe during NO-induced cardiac protection.