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Studies on the mechanism of flagellar movement using mammalian seminal plasma motility inhibitor protein.

利用哺乳动物精浆运动抑制蛋白研究鞭毛运动机制。

基本信息

批准号：
63540567
负责人：
OKUNO Makoto
金额：
$ 1.28万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

项目摘要

Mammalian seminal plasma contains a inhibitor to immobilize the demembranated flagellar movement. The inhibitor (SPMI, seminal plasma motility inhibitor) was prepared from the boar. The purified SPMI was consisted with three subunits, 14, 16 and 18 KD in SDSPAGE. SPMI inhibited not only mammalian but also fish and sea urchin sperm flagellar movement so that it seemed to inhibit the flagellar movement universally. When the demembranated sea urchin sperm flagella were exposed to SPMI the amplitude of bend decreased apparently, followed by irregular beat and then, stopped. The sliding disintegration of doublet microtubules were inhibited by SPMI, however, the end of flagella split out to individual microtubules and the bends were relaxed. ATPase activity was partially inhibited by SPMI. It was assumed that the amplitude of flagellar bend was closely related to the viscoelasticity in flagellar axoneme and SPMI increased it. It was also observed that the very slow bend propagation occurred in the presence of high concentration of SPMI. In that case, some wave parameters were preserved during the bend propagation, suggesting that these parameters should be very important to understanding the control mechanism of flagellar movement.Effects of poly amino acid on flagellar movement was studied. Only the basic and high molecular weight (15 KD or more) poly amino acids such as poly-L-lysine (PLL) inhibited the flagellar movement. The bend amplitude rather than the beat frequency was inhibited by PLL. Bends were relaxed even in the presence of high concentration of PLL. These results suggested that the inhibition manner of PLL was similar to that of SPMI in some parts. Therefore, it was likely that PLL increased the inter microtubule viscoelasticity.The microinjection of SPMI into the sea urchin blastomere was carried out. SPMI provided some inhibitory effects in a certain period prior to the cell division.

哺乳动物精浆中含有一种抑制脱膜鞭毛运动的抑制剂。抑制剂（SPMI，精浆运动抑制剂）从公猪制备。纯化的SPMI经SDS-PAGE分析由14、16和18 KD三个亚基组成。SPMI不仅抑制哺乳动物精子的鞭毛运动，而且也抑制鱼类和海胆精子的鞭毛运动，因此SPMI似乎具有普遍的抑制作用。去膜后的海胆精子鞭毛经SPMI处理后，弯曲幅度明显减小，随后出现不规则搏动，最后停止。SPMI抑制了微管的滑动解体，但鞭毛末端分裂成单个微管，弯曲松弛。ATP酶活性被SPMI部分抑制。推测鞭毛弯曲的幅度与鞭毛轴丝的粘弹性密切相关，而SPMI使鞭毛轴丝的粘弹性增大，并观察到在高浓度SPMI存在下，弯曲的传播非常缓慢。在这种情况下，一些波参数在弯曲传播过程中得以保留，这表明这些参数对于理解鞭毛运动的控制机制是非常重要的。只有碱性和高分子量（15 KD或更大）的聚氨基酸，如聚-L-赖氨酸（PLL）抑制鞭毛运动。PLL抑制的是弯曲幅度而不是拍频。即使在高浓度PLL的存在下，弯曲也是松弛的。这些结果表明，PLL的抑制方式与SPMI的抑制方式在某些部位相似。因此，PLL可能增加了微管间的粘弹性。SPMI在细胞分裂前的一段时间内具有一定的抑制作用。