STRUCTURE OF VERTEBRATE THICK ELEMENT

脊椎动物厚元件的结构

• 批准号: 5211925
• 负责人: ROBERT W KENSLER
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5211925
• 关键词: SDS polyacrylamide gel electrophoresis; chickens; computer simulation; crosslink; electron microscopy; goldfish; image processing; monoclonal antibody; muscle contraction; myosins; protein structure; stainings; striated muscles; western blottings

Skeletal muscle is the primary tissue involved in athe movements and production of force in man and all higher animals. Although the number of major pathologies affecting skeletal muscle are relatively few (multiple sclerosis, muscular dystrophy for example), the severity of these pathologies underlines the importance of muscle to the healthy individual, and th need to understand the normal functioning of this tissue. Contraction is known to involve an interaction between two sets of microscopic filaments; the myosin-containing thick filaments and the actin- containing thin filaments. In order to fully understand the contractile cycle it is essential to understand the structure of these filaments. The broad, long term objective of the research is to understand the structure to the contractile cycle. The specific aim of the proposed studies is to elucidate the molecular arrangement of myosin and C-protein (an accessory protein) in the vertebrate thick filament. The studies to accomplish this goal involve; (1) the use of a combination of electron microscopy and computer image analysis to compute a three-dimensional reconstruction of the arrangement of myosin and accessory proteins in filaments isolated from goldfish and chickens; and (2) the use of SDS gel electrophoresis and immunolabelling with antibodies to chicken C-protein to determine whether C-protein is still present on the isolated thick filaments. In addition, the potential of immunolabelling for C-protein in freeze-substituted muscle will be examined for its ability to determine the location of C-protein relative to the crossbridge array. A comparison of the structure of these filaments with the earlier results on the frog thick filament should yield significant insights into athe structure of the vertebrate thick filament.

骨骼肌是参与运动和运动的主要组织 在人类和所有高等动物中产生的力量。尽管这一数字 影响骨骼肌的主要病变相对较少(多个 硬化症，例如肌营养不良症)，这些疾病的严重程度 病理学强调了肌肉对健康个体的重要性， 这需要了解这种组织的正常功能。 已知收缩涉及两组 微细丝；含肌球蛋白的粗丝和肌动蛋白- 含有细丝的。为了充分了解可缩性 了解这些细丝的结构是很重要的。这个 这项研究的广泛、长期目标是了解其结构 进入收缩周期。拟议研究的具体目的是 阐明肌球蛋白和C蛋白(一种辅助蛋白)的分子排列 蛋白质)在脊椎动物的粗丝中。为实现这一目标而进行的研究 目标涉及；(1)结合使用电子显微镜和 计算机图像分析计算三维重建 日本血吸虫肌球蛋白及其附属蛋白在细丝中的排列 金鱼和鸡；以及(2)使用十二烷基硫酸钠凝胶电泳法和 鸡C蛋白抗体免疫标记检测鸡C蛋白 分离的粗丝上仍有C-蛋白存在。此外, C蛋白在冷冻替代肌肉中的免疫标记潜力 将被检查其确定C蛋白位置的能力 相对于交叉桥阵列。这两个词的结构比较 具有较早结果的青蛙粗丝应该会产生 对脊椎动物粗丝结构的重大洞察。