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

Julian Borejdo, A. Shepard, I. Akopova, W. Grudzinski, J. Malicka

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

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 ∼600 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 ∼1/2 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 ∼170 ms after the rigor cross-bridge first binds ATP.

Original languageEnglish
Pages (from-to)3912-3921
Number of pages10
JournalBiophysical Journal
Volume87
Issue number6
DOIs
StatePublished - Dec 2004

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