Paschal B M, King S M, Moss A G, Collins C A, Vallee R B, Witman G B
Cell Biology Group, Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545.
Nature. 1987;330(6149):672-4. doi: 10.1038/330672a0.
The inner and outer arms of the flagellar axoneme generate the forces needed for flagellar movement; these arms contain ATPases called dyneins. To date, there has been no method for studying the mechanochemical transducing activity of isolated dyneins. Recently, it was found that the brain microtubule-associated protein (MAP) 1C is a microtubule-activated ATPase with the structural and force-producing properties of dynein. MAP 1C translocates microtubules in an in vitro gliding assay, suggesting that such an assay could also be used with axonemal dyneins. Here, we demonstrate that outer-arm dynein isolated from sea urchin (Strongylocentrotus purpuratus) sperm and adsorbed to a glass coverslip can translocate calf-brain microtubules along the surface of the coverslip. Our results conclusively demonstrate that outer-arm dynein by itself is capable of generating shearing forces. The ability to examine the force-generating properties of flagellar dynein in vitro should greatly facilitate studies of the mechanism of action of this important mechanochemical transducer.
鞭毛轴丝的内臂和外臂产生鞭毛运动所需的力；这些臂含有一种称为动力蛋白的ATP酶。迄今为止，尚无研究分离出的动力蛋白的机械化学转导活性的方法。最近发现，脑微管相关蛋白（MAP）1C是一种微管激活的ATP酶，具有动力蛋白的结构和产生力的特性。在体外滑行试验中，MAP 1C能使微管移位，这表明该试验也可用于轴丝动力蛋白。在此，我们证明从海胆（紫球海胆）精子中分离并吸附在玻璃盖玻片上的外臂动力蛋白能够使小牛脑微管沿着盖玻片表面移位。我们的结果确凿地证明，外臂动力蛋白自身能够产生剪切力。在体外检测鞭毛动力蛋白产生力的特性的能力将极大地促进对这种重要的机械化学转导器作用机制的研究。
