INTERACTION OF ALDH WITH MITOCHONDRIAL IMPORT APPARATUS

描述（来自申请人的摘要）：与主要用于制备乙醇的乙醇不同， 在肝细胞质中氧化，乙醛在代谢过程中形成， 乙醇在线粒体的基质空间中被氧化， 醛脱氢酶 核编码酶在合成后， 在游离核糖体上，必须转移到线粒体中， 通过蛋白酶的作用加工成成熟蛋白质。 的 用于定位和处理的信息位于N端 氨基酸的延伸，称为前导序列。 的结构 在胶束环境中通过2DNMNR确定了先导物。 因为 前导序列的固有结构灵活性，尚不清楚是否 这是呈现给每一个靶点的结构， 肽。 这些是外膜，外膜 受体/转运蛋白和加工蛋白酶。 已有可能 使用抗原和 交联实验 对其他组件的搜索将是 持续期间内的 这些蛋白质将被克隆、表达并重组到 脂质体。 合成肽的相互作用对应于那些 将研究人、牛肉和大鼠肝醛脱氢酶， 与脂质体结合，重构的转运体/受体组分， 加工肽酶。 肽的结构和动力学，当 结合，将推导出从NMR光谱数据使用转移NOE 实验和弛豫率。 乙醇对线粒体的影响 将重新调查进口，以确定它是否抑制蛋白质进口 除了乙醛脱氢酶以及它如何抑制乙醛的输入 脱氢酶。 对乙醇和乙醛的全面了解 氧化需要了解酶学的所有方面， 包括基因调控，反应机制和靶向适当的 细胞器

DESCRIPTION (from Applicant's Abstract): Unlike ethanol which is primarily oxidized in liver cytosol, acetaldehyde formed during the metabolism of ethanol is oxidized in the matrix space of the mitochondria by the action of aldehyde dehydrogenase. The nuclear coded enzyme, after being synthesized on free ribosomes, must be translocated into the mitochondria where it is processed to the mature protein by the action of a protease. The information for targeting and processing is located in an N-terminal extension of amino acids, called a leader sequence. The structure of the leader has been determined by 2DNMNR in a micellar milieu. Because of the inherent structural flexibility of the leader sequence, it is not known if this is the structure that is presented to each of the targets that binds the peptide. These are the outer membrane, the outer membrane receptor/translocator and the processing protease. It has been possible to identify some components of the import machinery using antigenic and cross-linking experiments. The search for additional components will be continued. The proteins will be cloned, expressed and reconstituted into liposomes. The interaction of synthetic peptides corresponding to those from human, beef, and rat liver aldehyde dehydrogenase will be studied when bound to the liposomes, reconstituted translocator/receptor components and processing peptidase. The structure and dynamics of the peptides, when bound, will be deduced from NMR spectroscopic data using transferred NOE experiments and relaxation rates. The effect of ethanol on mitochondrial import will be reinvestigated to determine if it inhibits import of proteins other than aldehyde dehydrogenase and how it inhibits the import of aldehyde dehydrogenase. A complete understanding of ethanol and acetaldehyde oxidation required that all aspects of the enzymology be understood, including gene regulation, reaction mechanisms and targeting to the proper organelle.