Rivolta M N, Urrutia R, Kachar B
Laboratory of Cellular Biology, National Institute on Deafness and other Communication Disorders, National Institutes of Health, Rockville, MD 20850, USA.
Biochim Biophys Acta. 1995 Nov 21;1232(1-2):1-4. doi: 10.1016/0005-2728(95)00107-1.
In the streaming cytoplasm of the Characean algae cell, the movement of organelles along actin bundles occurs at a striking rate of up to 60 microns s-1. To further characterize the molecular mechanisms responsible for this phenomenon, we have reconstituted the movement of actin filaments in vitro using defined biochemical components. We report that only a soluble cytoplasmic fraction devoid of organelles and filamentous material supports the movement of fluorescent-labeled actin filaments on glass at a rate of up to 60 microns s-1. This fraction also contains the K(+)-EDTA ATPase and the actin-activated Mg2+ ATPase activities characteristic of myosin proteins. Therefore, on the basis of these observations, we conclude that Nitella cells have a soluble pool of non-filamentous myosin molecules with the mechanochemical properties expected for a motor responsible for cytoplasmic streaming in vivo. The preparation and conditions described here should be useful for the purification of this translocator.
在轮藻细胞流动的细胞质中,细胞器沿着肌动蛋白束移动的速度惊人,可达60微米每秒。为了进一步阐明造成这种现象的分子机制,我们利用特定的生化成分在体外重建了肌动蛋白丝的移动。我们报告称,只有不含细胞器和丝状物质的可溶性细胞质部分能支持荧光标记的肌动蛋白丝在玻璃上以高达60微米每秒的速度移动。该部分还含有肌球蛋白蛋白特有的K(+) - EDTA ATP酶和肌动蛋白激活的Mg2+ ATP酶活性。因此,基于这些观察结果,我们得出结论:丽藻细胞具有可溶性的非丝状肌球蛋白分子库,其机械化学性质符合体内负责细胞质流动的分子马达的预期。本文所述的制备方法和条件应有助于这种转运体的纯化。