LIPID & PROTEIN TRAFFIC & DYNAMICS IN EPITHELIAL CELLS

脂质体

• 批准号: 2143759
• 负责人: MICHAEL A EDIDIN
• 金额: $ 85.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-01 至 1997-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2143759
• 关键词: biological signal transduction; epithelium; lipid transport; membrane lipids; membrane proteins; protein transport

This program project aims to understand and define some of the pathways, mechanisms and signals involved in the sorting, traffic and localization of membrane lipids and proteins to their functional surface domains in polarized epithelial cells. Six highly interrelated projects served by 4 core units will address and compare the problems of molecular targeting and localization in a variety of polarized epithelial cells [MDCK (kidney tubule epithelium), Caco-2 and HT-29 (intestinal epithelium) and WIF12-1 (hepatocyte)] in vitro. These projects address: 1) The lateral organization of membrane proteins and the function of this organization in the sorting and targeting of newly synthesized proteins. 2) The amino acid sequences involved in determining the distribution of an apical (dipeptidylpeptidase-IV) and a basolateral (Na,K-ATPase) protein and of a protein (LEP-100) cycling between lysosomes and the plasma membrane via endosomes. The intracellular transport of these proteins in differential epithelial cells will also be compared. (3) Molecule and vesicle traffic in different epithelial cell types, and the perturbation of this traffic resulting from changes in lipid metabolism or in microtubule architecture. This project will include work on a newly-derived hepatocyte cell line WIF12-1. 4) The isolation and characterization of cis- and trans- Golgi complex membranes as a function of cell type and extent of polarity. 5) The distribution and transport of lipids. Special emphasis will be placed on the movement of fluorescent sphingolipid analogs along secretory pathways and the effects which alterations in cellular sphingolipids and cholesterol may have on lipid sorting and transport. 6) The organization of microtubules, the routes of biosynthetic and endocytic vesicle transport and the mechanoenzymes required for this transport. These six projects will be supported by cores for cell culture, electron microscopy and other imaging, fluorescence measurements of molecular behavior, and administration. The structure, central support and resources proposed here will result in a highly collaborative and interactive approach to the problems of the establishment and maintenance of polarity of epithelial cells.

该计划项目旨在了解和定义一些途径， 分类，流量和本地化涉及的机制和信号 膜脂质和蛋白质到其功能表面结构域中的蛋白质 极化上皮细胞。 六个高度相互关联的项目由4提供 核心单位将解决并比较分子靶向和 在各种极化上皮细胞中定位[MDCK（肾脏 小管上皮），CACO-2和HT-29（肠上皮）和WIF12-1-12-1 （肝细胞）]体外。 这些项目地址：1）横向 膜蛋白的组织和该组织的功能 新合成蛋白的分类和靶向。 2）氨基酸 确定顶端分布所涉及的序列 （二肽基肽酶-IV）和基底外侧（Na，K-ATPase）蛋白和A的 蛋白质（LEP-100）在溶酶体和质膜之间循环 内体。 这些蛋白质中的细胞内转运 还将比较上皮细胞。 （3）分子和囊泡流量 在不同的上皮细胞类型和此流量的扰动中 脂质代谢或微管体系结构的变化产生。 该项目将包括在新衍生的肝细胞细胞系上的工作 WIF12-1。 4）顺式和跨性别的隔离和表征 复杂的膜是细胞类型和极性程度的函数。 5） 脂质的分布和运输。 特别重点将放置 关于荧光鞘脂类似物沿分泌的运动 途径以及细胞鞘脂和 胆固醇可能具有脂质分类和运输。 6）组织 微管，生物合成和内吞囊泡的途径 和该运输所需的机械酶。 这六个项目将由细胞培养的核心支持电子 显微镜和其他成像，分子的荧光测量 行为和管理。 结构，中央支持和资源 这里提出的将导致高度协作和互动性 解决极性建立和维持问题的方法 上皮细胞。