Rotation of the lever arm of myosin in contracting skeletal muscle fiber measured by two-photon anisotropy

被引:5
|
作者
Borejdo, J
Shepard, A
Akopova, I
Grudzinski, W
Malicka, J
机构
[1] Univ N Texas, Dept Mol Biol & Immunol, Ft Worth, TX 76107 USA
[2] Univ Maryland, Sch Med, Dept Biochem & Mol Biol, Ctr Fluorescence Spect, Baltimore, MD 21201 USA
关键词
D O I
10.1529/biophysj.104.045450
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The rotation of the lever arm of myosin cross-bridges is believed to be responsible for muscle contraction. To resolve details of this rotation, it is necessary to observe a single cross-bridge. It is still impossible to do so in muscle fiber, but it is possible to investigate a small population of cross-bridges by simultaneously activating myosin in a femtoliter volume by rapid release of caged ATP. In earlier work, in which the number of observed cross-bridges was limited to similar to600 by confocal microscopy, we were able to measure the rates of cross-bridge detachment and rebinding. However, we were unable to resolve the power stroke. We speculated that the reason for this was that the number of observed cross-bridges was too large. In an attempt to decrease this number, we used two-photon microscopy which permitted observation of similar to1/2 as many cross-bridges as before with the same signal/noise ratio. With the two-photon excitation, the number of cross-bridges was small enough to resolve the beginning of the power stroke. The results indicated that the power stroke begins similar to170 ms after the rigor cross-bridge first binds ATP.
引用
收藏
页码:3912 / 3921
页数:10
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