INTERACTION OF ALDH WITH MITOCHONDRIAL IMPORT APPARATUS

描述(摘自申请者摘要)：与乙醇不同，乙醇主要是 在肝脏胞浆中氧化，在代谢过程中生成乙醛 乙醇在线粒体的基质空间被氧化，其作用是 乙醛脱氢酶。核编码酶，合成后 在游离核糖体上，必须移位到它所在的线粒体 通过蛋白酶的作用加工成成熟的蛋白质。这个 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，用2DNMNR测定了先导数。因为 先导序列固有的结构灵活性，目前尚不清楚 这是提供给绑定的每个目标的结构 多肽。这些是外膜，外膜 受体/转运体和加工酶。已经有可能 使用抗原性和抗原性识别进口机械的一些组件 交联性实验。对其他组件的搜索将是 继续。这些蛋白质将被克隆、表达并重组到 脂质体。与之相对应的合成肽的相互作用 来自人、牛肉和大鼠肝脏的乙醛脱氢酶将在何时进行研究 结合到脂质体、重组转运体/受体组件和 加工多肽酶。多肽的结构和动力学，当 界限，将从使用转移的NOE的核磁共振波谱数据推断 实验和松弛速率。乙醇对线粒体的影响 进口将被重新调查，以确定它是否抑制蛋白质的进口 除了乙醛脱氢酶之外，它是如何抑制乙醛的进口的 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.