STRUCTURE AND FUNCTION OF MEMBRANE TRANSPORT PROTEINS

膜运输蛋白的结构和功能

• 批准号: 3227627
• 负责人: DANIEL S LEVY
• 金额: $ 14.51万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-08-01 至 1987-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3227627
• 关键词: affinity labeling; anions; carbohydrate structure; chemical fingerprinting; hepatocellular carcinoma; ion transport; liposomes; liver cells; membrane reconstitution /synthesis; membrane structure; phase change; photochemistry; proteolysis; radiotracer; sulfates; tissue /cell culture

This proposal seeks to characterize the membrane components involved in the hepatocyte anion transport system, and to elucidate their disposition within the membrane system with the ultimate goal of understanding the architecture, dynamics and regulation of transport channels. Studies using hepatocytes derived from resting and regenerating liver, and hepatoma tissue culture (HTC) cells will be concerned with the analysis of anion transport properties and the identification, isolation and characterization of the membrane associated anion transport proteins using photoaffinity labeling in conjunction wwth differential extraction and chromatographic techniques. The purified proteins will be characterized by amino acid and carbohydrate analysis as well as by tryptic fingerprint mapping. The functional role of these components in the transport process will be evaluated using liposome reconstitution procedures. The proteoliposome system will be evaluated by characterizing sulfate transport properties and the effects of anion transport inhibitors. The structural organization and spacial orientation of the transport components in the hepatocyte and proteoliposome systems will be characterized using a variety of photoaffinity probes in conjunction with proteolytic dissection. The effect of proteolysis of the membrane assocated transport proteins on the transport function of the cells and liposomes will also be evaluated. These studies should provide many insights into the mechanism of hepatocyte anion transport and contribute to our general understanding of transmembrane transport channels.

该提案旨在表征涉及的膜组件， 肝细胞阴离子转运系统，并阐明其在肝细胞内的分布。 膜系统的最终目标是理解建筑， 运输渠道的动态和监管。 使用肝细胞的研究 来源于静息和再生肝脏，以及肝癌组织培养物（HTC） 细胞将关注阴离子传输特性的分析， 膜缔合阴离子的鉴定、分离和表征 利用光亲和标记结合差异表达的转运蛋白 萃取和色谱技术。 纯化的蛋白质将被 通过氨基酸和碳水化合物分析以及胰蛋白酶 指纹图谱 这些成分在运输中的功能作用 将使用脂质体复溶程序评价工艺。 的 将通过表征硫酸盐转运来评价蛋白脂质体系统 阴离子转运抑制剂的性质和作用。 结构性 运输组成部分的组织和空间定位， 肝细胞和蛋白脂质体系统将使用各种 光亲和探针结合蛋白水解裂解。 的影响 膜相关转运蛋白的蛋白水解作用 还将评价细胞和脂质体的功能。 这些研究 为肝细胞阴离子转运机制的研究提供了新的思路 并有助于我们对跨膜转运通道的一般理解。