STRUCTURE/FUNCTION OF THE GOLGI MEMBRANE SKELETON

高尔基膜骨架的结构/功能

• 批准号: 6520018
• 负责人: KENNETH A BECK
• 金额: $ 15.8万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6520018
• 关键词: Golgi apparatus; MDCK cell; SDS polyacrylamide gel electrophoresis; ankyrins; binding proteins; electron microscopy; fluorescence microscopy; genetic library; immunoelectron microscopy; light microscopy; membrane activity; membrane structure; molecular cloning; northern blottings; nucleic acid sequence; photomicrography; polymerase chain reaction; protein biosynthesis; protein isoforms; protein protein interaction; protein structure function; protein transport; spectrin; tissue /cell culture; yeast two hybrid system

The spectrin membrane skeleton is a ubiquitous, submembranous cytoskeleton that associates with the cytoplasmic surface of membranes. Genetic defects and deficiencies in erythrocyte isoforms of membrane skeleton components give rise to hemolytic disorders in humans. Two functions have been identified for the spectrin membrane skeleton: the maintenance of plasma membrane structural integrity and the formation of discrete membrane domains enriched in classes of membrane proteins. The recent identification of Golgi localized isoforms of membrane skeleton proteins reveals an even broader potential role for the membrane skeleton in the regulation of cellular architecture. The goal of this proposal is to characterize this Golgi membrane skeleton and to determine its function. It is hypothesized that the Golgi isoform of the membrane skeleton serves two critical functions which are analogous to the functions proposed for the plasma membrane isoforms: the maintenance of Golgi membrane structural identify and the formation of distinct membranes involved in membrane protein sorting within the Golgi complex. The specific questions that will be addressed are: 1. Are Golgi membrane skeleton proteins unique isoforms specialized for Golgi function? The precise identify of Golgi membrane skeleton proteins spectrin and ankyrin will be identified by molecular cloning. These studies will be essential for subsequent aims. 2. Will the identification of membrane skeleton bind proteins reveal critical features of the structure and function of the Golgi membrane skeleton? It is proposed that cytoplasmic binding proteins will be regulators of membrane skeleton assembly and integral membrane proteins that interact with the membrane skeleton assembly and integral membrane proteins that interact with the membrane skeleton will be "substrate proteins" acted on my membrane skeleton. The identification of these proteins will therefore provide important information regarding the assembly of the membrane skeleton as well as its protein. 3. Will dominant negative perturbations of the Golgi membrane skeleton illustrate a role for the Golgi membrane skeleton in the maintenance of Golgi structure and the formation of distinct membrane domains? Dominant negative disruption of the Golgi membrane skeleton will be performed in intact cells to directly test the proposed functions of the novel cytoskeletal structure.

光谱膜骨骼是一种无处不在的，肿瘤的细胞骨架，与膜的细胞质表面相关。膜骨骼成分的红细胞同工型中的遗传缺陷和缺陷会导致人类溶血性疾病。已经确定了光谱膜骨骼的两个功能：维持质膜结构完整性以及富含膜蛋白类别的离散膜结构域的形成。膜骨骼蛋白的高尔基体局部同工型的最新鉴定揭示了膜骨架在调节细胞结构中的潜在作用。该建议的目的是表征该高尔基膜骨架并确定其功能。假设膜骨架的高尔基同工型具有两个关键功能，这些功能类似于质膜同工型提出的功能：维持高尔基膜结构鉴定和与莫布雷蛋白在高尔基体内膜蛋白分类相关的膜的形成。将要解决的具体问题是：1。高尔基膜骨架蛋白是专门用于高尔基体功能的独特同工型吗？高尔基膜骨骼蛋白质谱和斜肌的精确识别将通过分子克隆鉴定。这些研究对于后续目标至关重要。 2。识别膜骨骼结合蛋白会揭示高尔基膜骨骼的结构和功能的关键特征吗？有人提出，细胞质结合蛋白将是与膜骨架组件相互作用的膜骨架组件和整体膜蛋白的调节剂，与膜骨骼相互作用的整合膜蛋白相互作用将是“底物蛋白”在我的膜骨架上作用于“底物蛋白”。因此，这些蛋白质的鉴定将提供有关膜骨骼及其蛋白质组装的重要信息。 3。高尔基膜骨架的主要负扰动会说明高尔基膜骨架在维持高尔基体结构和形成不同膜域中的作用吗？高尔基膜骨骼的主要负面破坏将在完整的细胞中进行，以直接测试新型细胞骨架结构的拟议功能。