LATERAL ORGANIZATION OF EPITHELIAL CELL SURFACES

上皮细胞表面的横向组织

• 批准号: 6564272
• 负责人: MICHAEL A EDIDIN
• 金额: $ 14.67万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6564272
• 关键词: Golgi apparatus; HMG coA reductases; MDCK cell; apical membrane; basolateral membrane; biological signal transduction; cell sorting; cellular polarity; chimeric proteins; epithelium; fluorescence resonance energy transfer; fluorimetry; glycoproteins; glycosphingolipids; glycosylphosphatidylinositols; green fluorescent proteins; liposomes; liver cells; low density lipoprotein; membrane activity; membrane lipids; membrane proteins; oxidoreductase inhibitor; photochemistry; protein transport

Epithelial cells, which comprise 50% of all cells in the body [and account for 90% of all human cancers] have as a defining function the creation and maintenance of physiological compartments of the body. They are functionally and morphologically polarized in order to perform this function. Lipid-anchored proteins, GPI=proteins and glycosphingolipids are concentrated at the apical surfaces of most polarized epithelial cells. We propose to use biophysical techniques to study the formation, and organization of the sorting units for GPI-proteins in the Golgi and plasma membranes and to compare the sorting units in MDCK cells, in which GPI-proteins are delivered directly to the apical surface, and in WIF-B hepatocytes, in which apical proteins take an indirect route to the apical surface. We will also compare the lateral organization of different GPI-proteins in MDCK cells and in liposomes. The following questions will be address: 1) How are GPI-proteins and GSL concentrated and segregated from other proteins in the Golgi? 2) What are the cell & protein specificities of the clustering and polarity of GPI-proteins in cells grown in complete medium and abolish their polarity in cells deprived of LDL? Our techniques to address these questions include fluorescence photo bleaching and recovery, FPR, fluorescence loss in photo bleaching, FLIP, and imaging fluorescence resonance energy transfer, FRET.

上皮细胞，占体内所有细胞的50％[和 占所有人类癌的90％]作为定义功能 人体生理隔室的创建和维护。 它们在功能和形态上是极化的，以便 执行此功能。 脂质锚定蛋白，GPI =蛋白质和 糖磷脂集中在大多数的顶部表面 极化上皮细胞。 我们建议使用生物物理 研究分类的形成和组织的技术 GOLGI和质膜中GPI蛋白的单位 比较MDCK单元中的分类单元，其中GPI-蛋白是 直接传递到顶部表面，并在Wif-B肝细胞中传递 顶端蛋白采取间接路线到达顶端 表面。 我们还将比较不同的横向组织 MDCK细胞和脂质体中的GPI-蛋白。 以下问题 将在地址：1）GPI-蛋白和GSL如何集中 与高尔基体中的其他蛋白质分离？ 2）什么是细胞＆ GPI蛋白的聚类和极性的蛋白质特异性 在完整培养基中生长的细胞中，废除其在细胞中的极性 被剥夺了LDL？ 我们解决这些问题的技术包括 荧光照片漂白和恢复，FPR，荧光丧失 照片漂白，翻转和成像荧光共振能量 转移，fret。