Arner A
Pflugers Arch. 1983 Apr;397(1):6-12. doi: 10.1007/BF00585160.
Rat portal veins were chemically skinned using Triton X-100 and mounted for isometric and quick release experiments at 20 degrees C. The skinned preparations were activated by Ca2+ (EGTA-buffered) in solutions containing 2mM free-Mg2+ and 1 microM calmodulin. Half maximal isometric force was obtained at pCa = 6.2. Maximal force of the skinned preparations, at pCa = 4.5, was 8.2 +/- 0.8 mN/mm2 (n = 6). Force-velocity relations were determined at varied Ca2+-concentrations. Maximal shortening velocity (Vmax) was 0.10 +/- 0.01 lengths/s at pCa = 4.5. At decreasing Ca2+-levels Vmax decreased (at pCa = 6.25, Vmax = 0.051/s). At pCa = 9 an increased level of free-Mg2+ (15 mM) induces a slow and submaximal increase in tension. Force velocity relations of Mg2+-induced contractures were not different from those of Ca2+-contractures of similar magnitude (pCa = 6.3). The results indicate that the degree of activation of the contractile system, as regulated by Ca2+ and Mg2+, influences the kinetic properties of the actomyosin interaction as well as the force development.
使用Triton X-100对大鼠门静脉进行化学去膜处理,并在20℃下安装用于等长和快速释放实验。去膜制剂在含有2mM游离镁离子和1μM钙调蛋白的溶液中由钙离子(EGTA缓冲)激活。在pCa = 6.2时获得半数最大等长力。在pCa = 4.5时,去膜制剂的最大力为8.2±0.8 mN/mm2(n = 6)。在不同钙离子浓度下测定力-速度关系。在pCa = 4.5时,最大缩短速度(Vmax)为0.10±0.01长度/秒。随着钙离子水平降低,Vmax降低(在pCa = 6.25时,Vmax = 0.051/秒)。在pCa = 9时,游离镁离子水平升高(15 mM)会导致张力缓慢且未达到最大值的增加。镁离子诱导的挛缩的力-速度关系与类似大小的钙离子挛缩(pCa = 6.3)的力-速度关系没有差异。结果表明,由钙离子和镁离子调节的收缩系统的激活程度会影响肌动球蛋白相互作用的动力学特性以及力的产生。