MOLECULAR MECHANISMS OF PEROXISOMAL PROTEIN IMPORT

描述（来自申请人摘要的逐字逐句）： 该实验室的目的是确定所需的生化成分， 阐明了人类过氧化物酶体输入它们的分子机制， 组成酶。过氧化物酶体是重要的细胞内细胞器， 大量的生化和代谢过程。的存在 破坏性的人类遗传疾病，其中过氧化物酶体未能输入其 基质酶最好地说明了细胞器的重要性， 人体生理学 绝大多数过氧化物酶体蛋白含有特异性靶向信号， 称为PTS 1，其用于将它们识别为输入底物。的基础 这种鉴定与PTS 1特异性受体分子有关， Pex5p。受体-底物复合物然后与受体的组分相互作用。 过氧化物酶体膜和易位包埋。尽管有许多分子， 称为过氧化物酶，涉及过氧化物酶体的各个方面 组装，详细描述基本分子机制 与过氧化物酶体蛋白输入相关的信息尚未公布。 我们了解过氧化物酶体蛋白输入的方法是 在体外重建该过程的各个方面。在本提案中，我们 将利用许多这样的测定来检查细胞内运输 Pex 5 p和PTS 1，以及获得对此的机械见解， 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Pex 5 p与PTS 1的结合。 2.分析Pex 5 p-PTS 1复合物在过氧化物酶体膜上的对接。 3.分析Pex 5 p和PTS 1的导入。 4.分析Pex 5 p回收。

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.