STRUCTURE OF VERTEBRATE THICK ELEMENT

脊椎动物厚元件的结构

• 批准号: 6271698
• 负责人: ROBERT W KENSLER
• 金额: $ 12.07万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6271698
• 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.

骨骼肌是参与骨骼运动的主要组织， 在人类和所有高等动物中产生力。 数量虽然 影响骨骼肌的主要病理相对较少（多种 硬化症，肌肉萎缩症例如），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， 我们需要了解这个组织的正常功能。 已知收缩涉及两组分子之间的相互作用。 显微镜下的肌丝;含肌球蛋白的粗丝和肌动蛋白， 含有细丝的。 为了充分了解收缩性 因此，了解这些细丝的结构是至关重要的。 的 研究的一个广泛的、长期的目标是了解结构 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（一种辅助蛋白）的分子排列 蛋白质）在脊椎动物的粗丝。 为了实现这一目标， 目标涉及;（1）使用电子显微镜和 计算机图像分析来计算 肌球蛋白和辅助蛋白在肌丝中的排列 金鱼和鸡;（2）使用SDS凝胶电泳， 用鸡C蛋白的抗体进行免疫标记， C蛋白仍然存在于分离的粗丝上。 此外，本发明还提供了一种方法， 冷冻替代肌肉中C蛋白免疫标记潜力 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 与较早的结果对青蛙粗丝应屈服 对脊椎动物粗丝的螺旋结构有重要的了解。