We have used a high-resolution small angle X-ray scattering system, together with a high-performance CCD camera, on the BioCAT beamline at the APS synchrotron radiation facility at the Argonne National Laboratory, to study X-ray interference effects in the meridional reflections generated by the arrays of myosin crossbridges in contracting muscle. These give information about axial movements of the myosin heads during contraction with sub-nanometer resolution. Using whole intact muscle preparations (frog sartorius) we have been able to record the detailed behavior of M3 (the first order meridional reflection from the myosin crossbridges, at 14.56 nm) at each of a number of quick releases of increasing magnitude, on the same specimen, and at the same time make similar measurements on higher order myosin meridional reflections, particularly M6. The latter provides information about the dispersion of lever arm angles of the actin-attached myosin heads. The observations show that in isometric contraction the lever arm angles are dispersed through +/- 20-25 degrees on either side of a mean orientation that is about 60 degrees away from their orientation at the end of the working stroke: and that they move towards that orientation in synchronized fashion, with constant dispersion, during quick releases. The relationship between the shift in the interference fringes (which measures the shift of the myosin heads scattering mass towards the center of the sarcomere, and the changes in the total intensity of the reflections, which measures the changes in the axial profile of the heads, is consistent with the tilting lever arm mechanism of muscle contraction. Significant fixed contributions to the meridional reflections come from unattached myosin heads and from backbone components of the myosin filaments, and the interaction of these with the contributions from actin-attached myosin heads determines the behavior of these reflections.

译文

:我们在阿贡国家实验室的APS同步加速器辐射设施的BioCAT光束线上使用了高分辨率小角度X射线散射系统以及高性能CCD相机,研究了X射线在X射线干扰方面的影响收缩肌中肌球蛋白横桥阵列产生的子午反射。这些以亚纳米分辨率给出有关收缩期间肌球蛋白头的轴向运动的信息。使用完整的完整肌肉制备物(青蛙sartorius),我们能够在相同的数量的多个快速释放中,分别记录M3的详细行为(来自肌球蛋白交叉桥的一阶子午反射,在14.56 nm处)。标本,同时对高阶肌球蛋白子午线反射(特别是M6)进行类似的测量。后者提供了有关附有肌动蛋白的肌球蛋白头的杠杆臂角度分散的信息。观察结果表明,在等距收缩中,杠杆臂角度在平均方向的任一侧上通过20-25度分散,该平均方向与工作行程结束时的方向相差约60度:并且它们朝着该方向移动在快速释放过程中,以同步方式进行定位,并保持恒定的分散。干涉条纹的偏移(用于测量肌球蛋白头部散射质量向肌小节中心的偏移)与反射总强度的变化(用于测量头部的轴向轮廓的变化)之间的关系,与倾斜杠杆臂的肌肉收缩机制相一致,对子午反射的重要固定贡献来自未连接的肌球蛋白头和肌球蛋白丝的骨架成分,这些相互作用与肌动蛋白连接的肌球蛋白头的作用共同决定了行为。这些思考。

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