CORRELATION OF MUSCLE PHYSIOLOGY AND BIOCHEMISTRY

这项研究的目的是将生理，结构和 通过测量蛋白质来研究肌肉收缩的生化理论 运动和生化速率常数的肌动球蛋白ATP酶， 生理活性的骨骼和心肌制剂。 蛋白 在肌肉的电子显微照片中可以检测到结构变化 纤维超快速冻结在开始的张力发展后， 通过笼状ATP或笼状Ca 2+的激光脉冲光解激活。 笼 分子是惰性的，但是是底物或信号传导的光不稳定前体 分子，其在通过以下脉冲照射时释放活性化合物： 近紫外激光。 蛋白质旋转运动将使用 一种新的时间分辨荧光偏振方法， 微观空间分辨率 荧光肌动蛋白和肌球蛋白光 链将被显微注射到活的、分离的心脏或培养的 骨骼肌细胞 标记的肌动蛋白或肌球蛋白光的方向 将在自发或 电刺激收缩。 具体反应步骤如下： ATP酶机制将在工作肌纤维中进行研究， 测量溶剂和溶剂之间的稳定氧同位素的交换， 底物中的磷酸基团和ATP酶反应的产物。 同位素的分布将由气相色谱法测定 与磷酸盐化合物的质谱联用。 该氧 交换技术探讨了基本的生化反应， 完整的纤维晶格。 长丝滑动对这些的影响 将确定反应步骤。 该项目的结果将大大促进对 收缩过程，从而带来了更好的理解，这两个正常的 骨骼肌和心肌的病理状态以及其他类型的 细胞死亡率

The goal of this research is to relate physiological, structural and biochemical theories of muscle contraction by measurement of protein motions and biochemical rate constants of the actomyosin ATPase in physiologically active skeletal and heart muscle preparations. Protein structural changes will be detected in electron micrographs of muscle fibers ultra-rapidly frozen during the onset of tension development after activation by laser pulse photolysis of caged ATP or caged Ca2+. Caged molecules are inert, but photolabile precursors of substrate or signaling molecules that release the active compound on irradiation by a pulse of near-u.v. laser light. Protein rotational motions will be measured using a novel time-resolved fluorescence polarization method that will allow microscopic spatial resolution. Fluorescent actin and myosin light chains will be micro-injected into live, isolated cardiac or cultured skeletal myocytes. The orientation of the labeled actin or myosin light chains will be monitored during the time course of spontaneous or electrically stimulated contractions. Specific reaction steps of the ATPase mechanism will be investigated in working muscle fibers by measuring exchange of stable oxygen isotopes between the solvent and phosphate groups in the substrate and products of the ATPase reaction. The distribution of isotopes will be determined by gas chromatography coupled to mass spectrometry of the phosphate compounds. This oxygen exchange techniques probes the elementary biochemical reactions in the intact filament lattice. The influence of filament sliding on these reaction steps will be determined. Results from this project should significantly advance knowledge of the contractile process and thus bring a greater understanding of both normal and pathological states of skeletal and heart muscle and other types of cell mortality.