Stebbings H, Sharma K K
Department of Biological Sciences, Washington Singer Laboratories, University of Exeter, UK.
J Cell Sci. 1989 Jan;92 ( Pt 1):21-7. doi: 10.1242/jcs.92.1.21.
Microtubule-based translocation channels within the ovaries of the hemipteran insect Notonecta have been isolated by microdissection, and then detergent-extracted to leave a bundle of some 30,000 aligned microtubules. On addition of ATP and other hydrolysable nucleotides the microtubule bundle contorts into a helical configuration, a property we have called 'corkscrewing', before straightening again. This we believe to be indicative of force generation within the bundle. Electrophoretic analysis of the bundle of native microtubules reveals many polypeptides apart from the tubulins, and a number of these comigrate with microtubule-associated proteins (MAPs), which copolymerize with tubulins in reassembled microtubules from the same system. Corkscrewing does not occur if the microtubule bundle is pretreated with salt, a procedure that removes MAPs from microtubules, suggesting that the force is generated by a MAP or MAPs. In addition, certain minor polypeptides comprising the native microtubules are ATP-sensitive, a property expected of a microtubule motor.
半翅目昆虫仰泳蝽卵巢内基于微管的转运通道已通过显微切割分离出来,然后用去污剂提取,留下一束约30000条排列整齐的微管。加入ATP和其他可水解核苷酸后,微管束会扭曲成螺旋状结构,我们将这种特性称为“螺旋化”,之后又会再次变直。我们认为这表明微管束内产生了力。对天然微管束的电泳分析显示,除了微管蛋白外还有许多多肽,其中一些与微管相关蛋白(MAPs)迁移率相同,这些蛋白在来自同一系统的重组微管中与微管蛋白共聚。如果微管束用盐预处理,就不会发生螺旋化,盐处理会从微管中去除MAPs,这表明力是由一种或多种MAPs产生的。此外,构成天然微管的某些次要多肽对ATP敏感,这是微管马达所具有的特性。