Molecular mechanisms of the regulation of flagellar and cilliary movements.

Takahashi表明，海胆精子鞭毛轴丝的滑动速度，用弹性蛋白酶处理并在100 μ M ATP下重新激活，在高和低钙浓度下表现出不同。跳动的海胆精子鞭毛的表观滑动速度随着跳动频率的变化而变化，表明滑动速度不取决于ATP浓度（Shingyoji）。为了研究马达蛋白的分子机制，Toyoshima开发了一种新的体外运动分析系统，使我们能够将马达蛋白固定在特定位点。Ogawa建立了在BaculoGold系统中表达的编码动力蛋白马达结构域的cDNA。神谷分离出了新的衣原体突变体，缺乏不同的动力蛋白重链，从而明确了它们的功能。Mohri对各种动物鞭毛和纤毛外臂动力蛋白的分子形态进行了研究，发现原生生物的动力蛋白是三头的，而其他动物的动力蛋白是双头的。使用糜蛋白酶消化的电泳分析表明，α-微管蛋白亚型（α 5）位于四膜虫轴丝（中村）中的动力蛋白臂附近。据稻叶说，蛋白酶体位于鞭毛轴丝向质膜延伸的突起处。关于鞭毛运动的调控，Morisawa进行了相关蛋白的纯化，然后克隆了精子运动的启动、激活和精子趋化性的cDNA。Okuno证明哺乳动物精子运动的激活与微管滑动速度的变化、微管之间的相互抑制和蛋白质磷酸化密切相关。Nakaoka表明，草履虫纤毛皮质片中的异时波在含有环鸟苷酸的再激活培养基中重建。环磷酸腺苷依赖的29 kDa和65 kDa磷酸化对于控制草履虫纤毛搏动方向至关重要（

Takahashi showed that sliding velocities of sea urchin sperm flagellar axonemes, which were treated with elastase and reactivated at 100 muM ATP, were shown to be different in high and low calcium concentrations. Apparent sliding velocity of beating sea urchin sperm flagella was changed with changing the beat frequency, indicating that the sliding velocity is not determined with ATP concentration (Shingyoji). To study the molecular mechanism of motor proteins, Toyoshima has developed a novel in vitro motility assay system that allows us to hold motor proteins at specific site. Ogawa established to produce cDNA encoding the plausible dynein motor domain which is expressed in BaculoGold system. Kamiya isolated new Chlamydomonas mutants lacking different dynein heavy chains and thereby made clear their functions. Molecular shape of the outer arm dynein in flagella and cilia was surveyed among various animals by Mohri who showed the dynein in protists was three-headed, whereas that in other animals was two-headed. Electrophoretic analysis using chymotrypsin digestion has suggested that the alpha-tubulin isoform (a5) is located near dynein arms in Tetrahymena axonemes (Nakamura). According to Inaba, proteasomes are located at the projection extending from flagellar axoneme toward plasma membrance. Concerning the regulation of flagellar movements, Morisawa carried out purification of related proteins and then cDNA cloning in regard to the initiation, activation of sperm motility and chemotaxisof spermatozoa. Okuno demonstrated that the activation of mammalian sperm motility is tightry coupled with the changes in sliding velocity of microtubules, restaint between the microtubules and protein phosphorylation. Nakaoka showed that metachronal waves in ciliated cortical sheet of Paramecium was reconstituted in the reactivation medium containing cGMP.Cyclic AMP dependent phosphorylation of 29kDa and 65kDa were essential for controlling ciliary beating direction in Paramecium (