Yamashita H, Sugiura S, Sata M, Serizawa T, Iizuka M, Shimmen T, Momomura S
Second Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Mol Cell Biochem. 1993 Feb 17;119(1-2):79-88. doi: 10.1007/BF00926857.
We measured the relative sliding velocity of cardiomyopathic hamster cardiac myosin on actin cables by using an in vitro motility assay system. We also investigated the relationship between the velocity and both myosin isozyme content and ATPase activity. Cardiac myosin was obtained from cardiomyopathic hamsters (BIO 14.6; B) aged 3, 6, 9, and 18 months and age-matched controls (F1B; F). Long well-organized actin cables of an alga, Nitellopsis, were used for the motility assay. Small latex beads (2 microns in diameter) were coated with purified cardiac myosin. When myosin-coated beads were introduced into an algal cell in the presence of Mg-ATP, myosin interacted with actin and dragged the beads. Active movement of the beads along the actin cables was observed under a photomicroscope and the velocity was measured. The velocity was significantly lower in B than in F for each age group (0.47 vs. 0.71 microns/s at the age of 3 months, p < 0.05; 0.44 vs. 0.88 microns/s at 6 months, p < 0.01; 0.44 vs. 0.67 microns/s at 9 months, p < 0.01; 0.35 vs. 0.52 microns/s at 18 months, p < 0.05). Both Ca(2+)-activated ATPase activity and the percentage of alpha-myosin heavy chain were also lower in B than in F for each age group. When examined for individual specimens, there was a positive correlation between the velocity and both myosin Ca(2+)-activated ATPase activity (r = 0.84) and percentage of alpha-myosin heavy chain (r = 0.83). These data points of both control and cardiomyopathic hamsters were distributed near the regression line obtained from control and thyroxine-treated rabbits reported previously. The present results indicate that the difference in mechanical properties between control and cardiomyopathic cardiac myosin is attributed to isozyme redistribution and not to a qualitative change in each myosin molecule.
我们使用体外运动分析系统测量了心肌病仓鼠心肌肌球蛋白在肌动蛋白丝上的相对滑动速度。我们还研究了该速度与肌球蛋白同工酶含量及ATP酶活性之间的关系。从3、6、9和18月龄的心肌病仓鼠(BIO 14.6;B)以及年龄匹配的对照仓鼠(F1B;F)中获取心肌肌球蛋白。用一种藻类——轮藻的排列良好的长肌动蛋白丝进行运动分析。将小的乳胶珠(直径2微米)用纯化的心肌肌球蛋白包被。当在Mg-ATP存在的情况下将包被有肌球蛋白的珠子引入藻类细胞时,肌球蛋白与肌动蛋白相互作用并拖动珠子。在光学显微镜下观察到珠子沿肌动蛋白丝的活跃运动并测量速度。每个年龄组中,B组的速度显著低于F组(3月龄时为0.47对0.71微米/秒,p<0.05;6月龄时为0.44对0.88微米/秒,p<0.01;9月龄时为0.44对0.67微米/秒,p<0.01;18月龄时为0.35对0.52微米/秒,p<0.05)。每个年龄组中,B组的Ca(2+)激活的ATP酶活性和α-肌球蛋白重链的百分比也低于F组。对单个样本进行检测时,速度与肌球蛋白Ca(2+)激活的ATP酶活性(r = 0.84)以及α-肌球蛋白重链的百分比(r = 0.83)之间均呈正相关。对照仓鼠和心肌病仓鼠的这些数据点分布在先前报道的对照兔和甲状腺素处理兔所得的回归线附近。目前的结果表明,对照心肌肌球蛋白和心肌病心肌肌球蛋白之间力学性质的差异归因于同工酶的重新分布,而非每个肌球蛋白分子的质性变化。