Tawada K, Yoshida A, Morita K
J Biochem. 1976 Jul;80(1):121-7. doi: 10.1093/oxfordjournals.jbchem.a131243.
With the final aim of replacing myosin in a single muscle fiber, a technique for removing myosin almost completely from single fibers was developed and an attempt to "re-form" thick filaments in the myosin-free ghosts of single fibers was made. Complete removal of myosin from single glycerol-treated rabbit psoas fibers with Hasselbach-Schneider solution was difficult. However, when skinned glycerol-treated fibers were used and 1% (v/v) Triton X-100 was added to the Hasselbach-Schneider solution, almost complete removal of myosin was possible. The myosin-free ghosts of skinned single fibers were very fragile but retained the overall structure. In the ghost fibers, Z-membranes and thin filaments remained. The ghost fibers, after irrigation with myosin, underwent contraction upon addition of Mg-ATP. In the myosin-irrigated fibers, thick filaments were re-formed in lengths from one Z-membrane to the other Z-membrane of a sarcomere, running parallel to the thin filaments. The packing of these two filaments was not good. The isometric tension developed by the irrigated fibers upon addition of mg-ATP was about 10% of the tension developed by untreated fibers. The weak tension developed by irrigated fibers is probably due to the irregular packing of the thick and thin filaments in the fibers. The ghost fibers also contracted, though only slightly, upon addition of Mg-ATP after irrigation with heavy meromyosin.
以在单根肌纤维中替换肌球蛋白为最终目标，开发了一种从单根纤维中几乎完全去除肌球蛋白的技术，并尝试在单根纤维的无肌球蛋白的“幽灵”中“重新形成”粗肌丝。用哈塞尔巴赫 - 施奈德溶液从经甘油处理的兔腰大肌单根纤维中完全去除肌球蛋白很困难。然而，当使用去表皮的经甘油处理的纤维，并在哈塞尔巴赫 - 施奈德溶液中加入1%（体积/体积）的 Triton X - 100时，几乎可以完全去除肌球蛋白。去表皮单根纤维的无肌球蛋白“幽灵”非常脆弱，但保留了整体结构。在“幽灵”纤维中，Z线和细肌丝仍然存在。用肌球蛋白冲洗后的“幽灵”纤维，在加入Mg - ATP后会发生收缩。在经肌球蛋白冲洗的纤维中，粗肌丝在肌节的一个Z线到另一个Z线的长度范围内重新形成，与细肌丝平行排列。这两种肌丝的排列并不理想。经冲洗的纤维在加入mg - ATP后产生的等长张力约为未处理纤维产生张力的10%。经冲洗的纤维产生的弱张力可能是由于纤维中粗、细肌丝排列不规则所致。在用重酶解肌球蛋白冲洗后，“幽灵”纤维在加入Mg - ATP时也会收缩，不过收缩程度很小。
