MOLECULAR MECHANISMS OF PEROXISOMAL PROTEIN IMPORT

描述(逐字摘自申请者摘要)：长期目标 这个实验室是为了鉴定所需的生化成分和 阐明人类过氧化酶体导入其基因的分子机制 组成酶。过氧化物体是细胞内重要的细胞器，参与了 大量的生化和新陈代谢过程。的存在 一种毁灭性的人类遗传疾病，在这种疾病中，过氧化物体不能将其 基质酶最能说明细胞器在整体上的重要性 人类生理学。 绝大多数过氧化体蛋白都含有特定的靶向信号， 称为PTS1，用于识别它们是进口底物。的基础 这种鉴定与PTS1特异性受体分子有关， Pex5p。受体-底物复合体然后与 过氧化物体膜，并随后发生移位。尽管有一些分子， 被称为过氧化物素，与过氧酶体的各个方面有关 组件，详细描述了基本的分子机制 与过氧化物体相关的蛋白质进口尚不可用。 我们了解过氧酶体蛋白输入的方法是 在体外重建这一过程的各个方面。在这项提案中，我们 将利用一些这样的分析来检查细胞内的贩运 Pex5p和PTS1，以及获得对此的机械性见解 从根本上说重要的细胞过程。我们的多学科方法将 实现以下具体目标： 1.分析Pex5p与PTS1的结合。 2.分析Pex5p-PTS1复合体在过氧化体膜上的对接。 3.分析Pex5p和PTS1的进口情况。 4.分析Pex5p的回收利用。

DESCRIPTION (Verbatim from the applicant's abstract): The long term goal of this laboratory is to identify the required biochemical components and elucidate the molecular mechanisms by which human peroxisomes import their constituent enzymes. Peroxisomes are vital intracellular organelles involved in a vast array of biochemical and metabolic processes. The existence of devastating human genetic disorders in which the peroxisome fails to import its matrix enzymes best illustrates the importance of the organelle in overall human physiology. The vast majority of peroxisomal proteins contain a specific targeting signal, called PTS1, which serves to identify them as import substrates. The basis of this identification is an association with the PTS1-specific receptor molecule, Pex5p. Receptor-substrate complexes then interact with components of the peroxisome membrane, and translocation ensues. Although a number of molecules, called peroxins, have been implicated in various aspects of peroxisome assembly, a detailed description of the fundamental molecular mechanisms associated with peroxisomal protein import is not yet available. Our approach to gaining an understanding of peroxisomal protein import has been to reconstitute various aspects of the process in vitro. In this proposal, we will utilize a number of such assays to examine the intracellular trafficking of Pex5p and PTS1, as well as to gain mechanistic insights into this fundamentally important cellular process. Our multidisciplinary approach will address the following specific aims: 1. To analyze the binding of Pex5p to PTS1. 2. To analyze docking of the Pex5p-PTS1 complex at the peroxisome membrane. 3. To analyze import of Pex5p and PTS1. 4. To analyze Pex5p recycling.