MOLECULAR MECHANISMS OF PEROXISOMAL PROTEIN IMPORT

过氧化物酶体蛋白输入的分子机制

• 批准号: 6381622
• 负责人: Stanley Richard Terlecky
• 金额: $ 22.11万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6381622
• 关键词: adenosine triphosphate; biochemistry; biological signal transduction; cell membrane; cytoplasm; enzyme linked immunosorbent assay; fibroblasts; gel electrophoresis; heat shock proteins; human tissue; intracellular transport; laboratory rabbit; membrane permeability; membrane proteins; membrane reconstitution /synthesis; membrane transport proteins; molecular chaperones; molecular dynamics; peroxisome; protein binding; protein protein interaction; protein structure function; stoichiometry; tissue /cell culture

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.

描述（逐字病申请人的摘要）： 该实验室是要确定所需的生化成分和 阐明人过氧化物酶体导入的分子机制 组成酶。过氧化物酶体是参与的重要细胞内细胞器 一系列生化和代谢过程。存在 毁灭性的人类遗传疾病，其中过氧化物酶体未能进口 基质酶最能说明细胞器在总体上的重要性 人类生理学。 绝大多数过氧化物酶体蛋白包含特定的靶向信号， 称为PTS1，它可以将它们识别为导入基板。的基础 该鉴定是与PTS1特异性受体分子的关联 PEX5P。然后，受体 - 底物复合物与成分相互作用 过氧化物酶体膜和易位随之而来。虽然许多分子，但 称为过氧蛋白，已与过氧化物酶体的各个方面有关 组装，基本分子机制的详细描述 与过氧化物酶体蛋白进口相关。 我们了解过氧化物酶体蛋白进口的方法已经是 重建体外过程的各个方面。在这个建议中，我们 是否会利用许多此类测定法检查细胞内贩运 pex5p和pts1，以及为此获得机械洞察力 根本重要的细胞过程。我们的多学科方法将 解决以下特定目的： 1。分析PEX5P与PTS1的结合。 2。分析过氧化物酶体膜上PEX5P-PTS1复合物的对接。 3。分析PEX5P和PTS1的导入。 4。分析PEX5P回收。