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Journal of Cell Science, Vol 94, Issue 3 391-401, Copyright © 1989 by Company of Biologists
JOURNAL ARTICLES |
RW Kensler and M Stewart
Division of Structural and Systems Biology, School of Basic Life Sciences, University of Missouri, Kansas City 64108.
A procedure has been developed for isolating gold-fish skeletal muscle thick filaments that preserves the near-helical arrangement of the myosin cross-bridges under relaxing conditions. These filaments have been examined by electron microscopy and computer image analysis. Electron micrographs of the negatively stained filaments showed a clear periodicity associated with the crossbridges, with an axial repeat every 42.9 nm. Computed Fourier transforms of the negatively stained filaments showed a series of layer lines confirming this periodicity, and were similar to the X-ray diffraction patterns of fish muscle obtained by J. Hartford and J. Squire. Analysis of the computed transform data and filtered images of the isolated fish filaments demonstrated that the myosin crossbridges lie along three strands. Platinum shadowing demonstrated that the strands have a right-handed orientation, and computed transforms and filtered images of the shadowed filaments suggest that the crossbridges are perturbed both axially and azimuthally from an ideal helical arrangement.
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