CYTOSKELETAL INTERACTIONS OF DYSTROPHIN

肌营养不良蛋白的细胞骨架相互作用

• 批准号: 2081671
• 负责人: JAMES M ERVASTI
• 金额: $ 12.95万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-15 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2081671
• 关键词: actins; centrifugation; chemical binding; chickens; cytoskeleton; dystrophin; electron microscopy; glycoproteins; laboratory rabbit; light scattering; molecular pathology; muscular dystrophy; protein sequence; protein structure function

The long term objective of this research is to understand the structure and function of dystrophin, the protein product of the Duchenne muscular dystrophy gene. The strategy proposed to attain this goal is to identify the cytoskeletal proteins which interact with dystrophin as well as with the dystrophin-glycoprotein complex in rabbit skeletal muscle. Five specific aims are proposed: First, the F-actin binding properties of purified dystrophin-glycoprotein complex will be studied. A cosedimentation assay will be used to quantitatively define the binding of native dystrophin-glycoprotein complex to F-actin binding properties of dystrophin-glycoprotein complex that has been disrupted by various dissociative agents will be determined using the cosedimentation assay to determine whether any other components of the dystrophin-glycoprotein complex modulate the interaction between dystrophin and actin or bind actin independently of dystrophin. Third, the role of the dystrophin- glycoprotein complex in crosslinking F-actin into bundles of networks will be assessed by light scattering and low-g sedimentation assays, as well as by electron microscopy. These experiments will determine whether classification of dystrophin in the actin binding protein family is appropriate. Fourth, this project will use blot overlay and affinity precipitation techniques to determine whether components of the dystrophin-glycoprotein complex interact with other cytoskeletal proteins using. The complementary approaches outlined will yield important new information about the cytoskeletal interactions of dystrophin and its associated proteins. Thus, the proposed research will provide insight into the pathogenesis of Duchenne, Becker and possibly severe autosomal recessive muscular dystrophies through the definition of the function of dystrophin by its interactions with other proteins in normal muscle. The results of this research will also aid in the rational design of fully functional dystrophin mini-genes for use in dystrophin replacement therapies.

这项研究的长期目标是了解结构 Duchenne肌肉的蛋白质产物肌营养不良蛋白的功能 营养不良基因。提出实现此目标的策略是确定 与肌营养不良蛋白相互作用的细胞骨架蛋白 兔骨骼肌中的肌营养不良蛋白 - 糖蛋白复合物。五 提出了具体目的：首先，F-肌动蛋白结合特性 将研究纯化的肌营养不良蛋白糖蛋白复合物。一个 Cosseimentation Assay将用于定量定义结合 天然肌营养不良蛋白 - 糖蛋白复合物与F-肌动蛋白结合特性 肌营养不良蛋白 - 糖蛋白复合物已被各种破坏 解离剂将使用COSESIOMTIONS分析确定 确定肌营养不良蛋白 - 糖蛋白的其他成分是否 复杂调节肌营养素和肌动蛋白之间的相互作用或结合 肌动蛋白独立于肌营养物。第三，肌营养不良蛋白的作用 将F-肌动蛋白交联的网络束中的糖蛋白复合物 将通过光散射和低-G沉积测定法评估 以及通过电子显微镜。这些实验将确定是否 肌动蛋白结合蛋白家族中肌营养不良蛋白的分类为 合适的。第四，该项目将使用墨水叠加层和亲和力 降水技术以确定是否成分 肌营养不良蛋白 - 糖蛋白复合物与其他细胞骨架蛋白相互作用 使用。概述的互补方法将产生重要的新 有关肌营养不良蛋白及其的细胞骨架相互作用的信息 相关蛋白质。因此，拟议的研究将提供洞察力 进入Duchenne，Becker和可能严重常染色体的发病机理 隐性肌肉营养不良，通过定义 肌营养不良蛋白与正常肌肉中其他蛋白质的相互作用。这 这项研究的结果还将有助于完全设计 功能性肌营养不良蛋白微型基因用于肌营养不良蛋白替代 疗法。