Tsukita S, Tsukita S, Usukura J, Ishikawa H
J Cell Biol. 1983 May;96(5):1480-5. doi: 10.1083/jcb.96.5.1480.
With the rapid-freeze, deep-etch replica technique, the structural conformations of outer dynein arms in demembranated cilia from Tetrahymena were analyzed under two different conditions, i.e., in the absence of ATP and in the presence of ATP and vanadate. In the absence of ATP, the lateral view of axonemes was characterized by the egg-shaped outer dynein arms, which showed a slightly baseward tilt with a mean inclination of 11.1 degrees +/- 3.4 degrees SD from the perpendicular to the doublet microtubules. On the other hand, in the presence of 1 mM ATP and 100 microM vanadate, the outer arms were extended and slender and showed an increased baseward tilt with a mean inclination of 31.6 degrees +/- 4.9 degrees SD. In ATP-activated axonemes, these two types of arms coexisted, each type occurring in groups along one row of outer arms. These findings strongly suggest that the interdoublet sliding is caused by dynamic structural changes of dynein arms that follow the hydrolysis of ATP.
采用快速冷冻、深度蚀刻复型技术,在两种不同条件下,即无ATP以及有ATP和钒酸盐存在的情况下,分析了来自四膜虫的去膜纤毛中外动力蛋白臂的结构构象。在无ATP时,轴丝的侧视图显示为卵形的外动力蛋白臂,其从垂直于双联微管的方向向基部略有倾斜,平均倾斜度为11.1度±3.4度标准差。另一方面,在存在1 mM ATP和100 μM钒酸盐的情况下,外臂伸展且细长,基部倾斜度增加,平均倾斜度为31.6度±4.9度标准差。在ATP激活的轴丝中,这两种类型的臂共存,每种类型沿着一排外臂成组出现。这些发现有力地表明,双联微管间的滑动是由ATP水解后动力蛋白臂的动态结构变化引起的。