Hata H, Yano Y, Mohri T, Mohri H, Miki-Noumura T
J Cell Sci. 1980 Feb;41:331-40. doi: 10.1242/jcs.41.1.331.
We have prepared axonemes without outer dynein arms from sea-urchin (Pseudocentrotus depressus, Hemicentrotus pulcherrimus) sperm flagella by selective solubilization with NaCl. Electron microscopy revealed that the axonemes gradually lost their outer arms in 0.5 M NaCl during 10 min. Such axonemes retained 42.8 +/- 7.3% of their total axonemal ATPase activity and showed C, A, D and B bands in the dynein region of 4% SDS-gel, while a solubilized fraction of the outer arms consisted almost entirely of A band polypeptide. We have succeeded in causing extrusion of the outer doublets from such axonemes by addition of ATP and trypsin. A bundle of outer doublets was sometimes observed to be extruded first from an axoneme and to show bending motion for a while, subsequently followed by a sliding of separate doublets past each other. The speed of the tubule extrusion process was slower and around 60% of that of intact axonemes having both types of arm. These observations indicate that the inner arms have a function equivalent to that of the outer arms, of sliding on adjacent doublets, although the inner arms seem to be constituted from polypeptide(s) different from that of the outer arms.
我们通过用氯化钠选择性溶解，从海胆（凹形拟球海胆、美丽海胆）精子鞭毛中制备了没有外动力蛋白臂的轴丝。电子显微镜显示，轴丝在0.5M氯化钠中10分钟内逐渐失去其外臂。这种轴丝保留了其总轴丝ATP酶活性的42.8±7.3%，并在4%十二烷基硫酸钠凝胶的动力蛋白区域显示出C、A、D和B带，而外臂的可溶部分几乎完全由A带多肽组成。我们通过添加ATP和胰蛋白酶成功地使这些轴丝挤出外双联体。有时观察到一束外双联体首先从轴丝中挤出，并显示出一段时间的弯曲运动，随后是单独的双联体相互滑动。微管挤出过程的速度较慢，约为具有两种类型臂的完整轴丝速度的60%。这些观察结果表明，内臂具有与外臂相同的功能，即在相邻双联体上滑动，尽管内臂似乎由与外臂不同的多肽构成。
