Transient state kinetic analysis of the dynein ATPase.
Transient state kinetic analysis of the dynein ATPase.
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动力蛋白 ATP 酶的瞬态动力学分析。
DOI：
10.1002/cm.970020720
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Porter,ME
中科院分区：
文献类型：
--
作者：
Johnson,KA;Porter,ME
动力蛋白ATP酶途径的彻底分析不仅对于理解纤毛和鞭毛运动，而且对于理解微管相关ATP酶在产生其他细胞事件（如染色体运动、轴浆运输和膜结合囊泡的细胞内运动）的力中的潜在作用，都具有明显的重要性。迄今为止的工作已经确定，动力蛋白水解ATP [Gibbons，1963] 1，在不存在ATP的情况下，以通过加入ATP释放的刚性键交叉桥接纤毛外双重微管[Gibbons和Gibbons，1974; Zanetti，Mitchell和Warner，19791，并通过将跨桥作用与ATP的水解偶联而引起外部双联体微管的滑动[Summers和Gibbons，1971; Satir，1968; Brokaw and Benedict，1968; Gibbons and Gibbons，19721.一些研究者认为，动力蛋白跨桥循环可能与肌动球蛋白途径平行[销售和Gibbons，1979; Satir等人，1981; Okuno和Brokaw，1981 1]。这些研究依赖于与肌动球蛋白的类比，并使用假定的磷酸盐类似物钒酸盐来区分整个轴丝中ATP结合和ATP水解的作用。然而，钒酸盐的作用机制尚不清楚，并且动力蛋白动力学数据中缺少肌动球蛋白模型[Alcohn和Taylor，1971]的几个要素;即，没有直接测量ATP结合和水解以及ATP诱导的横桥释放的动力学;没有令人信服的证据表明微管蛋白在溶液中的生理条件下激活动力蛋白ATP酶;和AMPPNP，ATP的不可水解类似物，不能诱导精子鞭毛的僵直波松弛[Penningroth，Cheung和Olehnik，1980; Okuno和Brokaw，19811。
A thorough analysis of the dynein ATPase pathway is of obvious importance'to understanding not only ciliary and flagellar motility, but also the potential roles of microtubule-associated ATPases in producing force for other cellular events such as chromosome movement, axoplasmic transport, and the intracellular movements of membranebound vesicles. Work to date has established that dynein hydrolyses ATP [Gibbons, 19631, cross-bridges ciliary outer doublet microtubules in the absence of ATP in a rigortype bond that is released by the addition of ATP [Gibbons and Gibbons, 1974; Zanetti, Mitchell, and Warner, 19791, and causes the sliding of outer doublet microtubules by coupling cross-bridge action to the hydrolysis of ATP [Summers and Gibbons, 1971; Satir, 1968; Brokaw and Benedict, 1968; Gibbons and Gibbons, 19721. Several investigators have argued that the dynein cross-bridge cycle may parallel the actomyosin pathway [Sale and Gibbons, 1979; Satir et al, 1981; Okuno and Brokaw, 19811. These studies have relied upon analogies to actomyosin and upon the use of the presumptive phosphate analogue, vanadate, to distinguish effects due to ATP binding and ATP hydrolysis in the whole axoneme. However, the mechanism of action of vanadate is not known, and there are several elements of the actomyosin model [Lymn and Taylor, 19711 that are missing from the dynein kinetic data; namely, there has been no direct measurement of the kinetics of ATP binding and hydrolysis and of ATP-induced crossbridge release; there has been no convincing demonstration that tubulin activates the dynein ATPase under physiological conditions in solution; and AMPPNP, the nonhydrolyzable analogue of ATP, fails to induce the relaxation of rigor wave sperm flagella [Penningroth, Cheung, and Olehnik, 1980; Okuno and Brokaw, 19811.