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ATHE运动的主要组织， 人类和所有高等动物的力量产生。 虽然数量 影响骨骼肌的主要病理相对较少（多个 硬化症，肌肉营养不良），这些严重程度 病理强调了肌肉对健康个体的重要性， 并且需要了解该组织的正常功能。 已知收缩涉及两组之间的相互作用 微型细丝；含有肌球蛋白的厚细丝和肌动蛋白 - 含有细丝。 为了充分理解收缩 循环必须了解这些细丝的结构。 这 该研究的广泛，长期目标是了解结构 到收缩周期。 拟议研究的具体目的是 阐明肌球蛋白和C蛋白的分子排列（附件 蛋白质）在脊椎动物厚的细丝中。 实现这一目标的研究 目标涉及； （1）使用电子显微镜和 计算机图像分析以计算三维重建 肌球蛋白和辅助蛋白的排列在与丝质的细丝中隔离 金鱼和鸡； （2）使用SDS凝胶电泳和 与鸡C蛋白抗体的免疫标记，以确定是否是否 C蛋白仍然存在于孤立的厚细丝上。 此外， 冻结肌肉中C蛋白免疫标记的潜力 将检查其确定C蛋白位置的能力 相对于Crossbridge阵列。 这些结构的比较 青蛙厚丝上较早结果的细丝应产生 对脊椎动物厚细丝的体系结构的重要见解。