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ELLIPSOMETRY STUDIES OF CROSSBRIDGE MECHANISMS IN MUSCLE

肌肉跨桥机制的椭圆测量研究

基本信息

批准号：
3155453
负责人：
YIN YEH
金额：
$ 13.11万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
1980
资助国家：
美国
起止时间：
1980-04-01 至 1991-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3155453
关键词：
Anura birefringences conformation electron microscopy laser spectrometry muscle muscle contraction muscle proteins myofibrils myosins protein structure sarcomeres striated muscles

项目摘要

The studies outline in this proposal are directed towards providing additional insights into the mechanism of cross-bridge force generation in muscle cells. Non-invasive laser diffraction methods, with an emphasis on the changes of the optical polarization properties upon the interaction of light with the muscle fiber, is the technique chosen. The complete characterization of the optical polarization property known as the technique of ellipsometry, provides for the determination of both the phase and the amplitudes of the electric field components after its interaction. The amplitude information specifically singles out the intrinsically anisotropic elements of the basic muscle structural unit, the sarcomere. As such, time evolution studies of the changes in amplitudes monitor the orientational and intrinsic dynamics of anisotropic elements within the sarcomere. Predominently, these are the myosin rods: light meromyosin and subfragment 2. Since this region of the sarcomeric structure is undetected by x-ray diffraction methods and unable to a labelled by extrinsic probes, the ellipsometry technique provides an important complementary link. The phase information provides additional information about the contractile apparatus. Here, the phase shift measured in ellipsometry experiments contains both intrinsic as well as form contributions. In this proposal, the method of separation of intrinsic and form information, using simultaneously obtained amplitude data, is outlined. This additional separation of contributing parts gives this method added advantage over the technique of transmission birefringence, which also contain both anisotropic and phase information, but is inseparable. The sequence of activity outlined in this proposal will focus on: skinned fibers studies; myosin extracted, subfragment I reconstituted systems; and intact fibers under isometric tetanus and quick release conditions.

本提案中的研究大纲旨在提供对跨桥力产生机制的更多见解肌肉细胞。非侵入式激光衍射方法，重点是相互作用时光学偏振特性的变化光与肌肉纤维，是所选择的技术。完整的光学偏振特性的表征称为椭圆光度技术，提供了相位的确定以及相互作用后电场分量的振幅。幅度信息专门挑选出本质上基本肌肉结构单元肌节的各向异性元素。因此，振幅变化的时间演化研究监测各向异性元素的取向和内在动力学肌节。主要是肌球蛋白杆：轻色肌球蛋白和子片段 2。由于肌节结构的该区域未被检测到通过X射线衍射方法并且无法被外来探针标记，椭圆偏振技术提供了重要的补充环节。这相位信息提供了有关收缩的附加信息设备。这里，在椭圆光度实验中测量的相移包含内在贡献和形式贡献。在这个提案中，分离内在信息和形式信息的方法，使用概述了同时获得的幅度数据。这个额外的贡献部分的分离使该方法比传统方法具有更多优势透射双折射技术，其中还包含各向异性和相位信息可是密不可分的。的顺序为该提案中概述的活动将侧重于：皮纤维研究；提取肌球蛋白，亚片段I重构系统；和完整的纤维在等长破伤风和快速释放条件下。