THICK FILAMENT MYOSIN AND C-PROTEIN ARRANGEMENT

粗丝肌球蛋白和 C 蛋白排列

• 批准号: 3155803
• 负责人: Robert W. Kensler
• 金额: $ 8.4万
• 依托单位: ALLEGHENY UNIVERSITY OF HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-01-01 至 1988-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3155803
• 关键词: Anura; X ray crystallography; catfish; chickens; computer graphics /printing; electron microscopy; fresh water environment; gel electrophoresis; laboratory rabbit; monoclonal antibody; muscle function; muscle proteins; myofibrils; myosins; optics; protein structure; species difference; stainings; striated muscles

The arrangement of myosin and accessory protein such as C- protein in the thick filaments of fish, avian, and mammalian skeletal muscles is still not clearly elucidated. Recent work in our laboratory on the structure of frog thick filaments however, has demonstrated that the structure of vertebrate muscles may be amenable to analysis by a combination of electron microscopy, optical diffraction analysis, and computer image analysis. Additionally, it has been shown that thick filaments from fich, avian, and mammalian muscle can be isolated with a similarly well ordered array of crossbridges; thus making these filaments suitable for analysis by the same type of techniques which have proven useful for the frog thick filaments. The specific aim of the proposed research is to extend the techniques used for the frog thick filaments to the determination of the structure of the thick filaments of fish, avian, and mammalian skeletal muscles. The major objective of theses studies will be the determination of the arrangement of the myosin and accessory proteins in the filaments of these vertebrate classes. The studies to accomplish these aims will include: 1) the use of a combination of negative staining, platinum shadowing, rapid-freeze deep-etch cryo studies, and optical diffraction analysis to elucidate the major ultrastructural features of the filaments and to assess the preservation of the filaments as compared to the structural parameters expected from X-ray diffraction patterns of living muscle; 2) the use of computer image analysis of the filament images in the best electron micrographs to determine the rotational symmetry and arrangement of the myosin heads, and to produce a three-dimensional reconstruction of the structure of the filament; 3) the use of immunolabelling studies to determine the probable location of accessory proteins such as C-protein and X-protein on the filaments, and to correlate this information with the structure of the filament as seen in the three dimensional reconstructions. These studies will provide basic scientific information about the structure of muscle, and will thus provide a better foundation for understanding how pathological conditions may affect the functioning of this tissue.

肌球蛋白和辅助蛋白（如 C-）的排列 鱼类、鸟类和哺乳动物粗丝中的蛋白质 骨骼肌至今尚未明确阐明。 最近的工作于 然而，我们实验室对青蛙粗丝结构的研究， 已经证明脊椎动物的肌肉结构可能是 适合通过电子显微镜的组合进行分析， 光学衍射分析和计算机图像分析。 此外，已经表明，来自 Fich 的粗丝， 鸟类和哺乳动物的肌肉可以用类似的方法分离 井然有序的横桥阵列；从而使这些细丝 适用于通过相同类型的技术进行分析 事实证明，粗丝对青蛙很有用。 具体目标是 拟议的研究是为了扩展用于 青蛙粗丝的结构测定 鱼类、鸟类和哺乳动物骨骼肌的粗丝。 这些研究的主要目标是确定 肌球蛋白和辅助蛋白的排列 这些脊椎动物的细丝。 要完成的研究 这些目标将包括：1）使用负面的组合 染色、铂阴影、快速冷冻深蚀刻低温研究、 和光学衍射分析来阐明主要 细丝的超微结构特征并评估 与结构相比，细丝的保存 活体 X 射线衍射图谱预期参数 肌肉; 2）利用计算机图像分析灯丝 最好的电子显微照片中的图像以确定 肌球蛋白头的旋转对称性和排列，以及 产生结构的三维重建 灯丝； 3）利用免疫标记研究来确定 辅助蛋白（例如 C 蛋白）的可能位置 细丝上的 X 蛋白，并将此信息与 从三维图中看到的灯丝结构 重建。 这些研究将提供基本的科学依据 有关肌肉结构的信息，从而提供 更好地了解病理状况的基础 可能会影响该组织的功能。