Electron tomography of fast frozen, stretched rigor fibers reveals elastic distortions in the myosin crossbridges.

Published

Journal Article

As a first step toward freeze-trapping and 3-D modeling of the very rapid load-induced structural responses of active myosin heads, we explored the conformational range of longer lasting force-dependent changes in rigor crossbridges of insect flight muscle (IFM). Rigor IFM fibers were slam-frozen after ramp stretch (1000 ms) of 1-2% and freeze-substituted. Tomograms were calculated from tilt series of 30 nm longitudinal sections of Araldite-embedded fibers. Modified procedures of alignment and correspondence analysis grouped self-similar crossbridge forms into 16 class averages with 4.5 nm resolution, revealing actin protomers and myosin S2 segments of some crossbridges for the first time in muscle thin sections. Acto-S1 atomic models manually fitted to crossbridge density required a range of lever arm adjustments to match variably distorted rigor crossbridges. Some lever arms were unchanged compared with low tension rigor, while others were bent and displaced M-ward by up to 4.5 nm. The average displacement was 1.6 +/- 1.0 nm. "Map back" images that replaced each unaveraged 39 nm crossbridge motif by its class average showed an ordered mix of distorted and unaltered crossbridges distributed along the 116 nm repeat that reflects differences in rigor myosin head loading even before stretch.

Full Text

Duke Authors

Cited Authors

  • Liu, J; Reedy, MC; Goldman, YE; Franzini-Armstrong, C; Sasaki, H; Tregear, RT; Lucaveche, C; Winkler, H; Baumann, BAJ; Squire, JM; Irving, TC; Reedy, MK; Taylor, KA

Published Date

  • September 2004

Published In

Volume / Issue

  • 147 / 3

Start / End Page

  • 268 - 282

PubMed ID

  • 15450296

Pubmed Central ID

  • 15450296

Electronic International Standard Serial Number (EISSN)

  • 1095-8657

International Standard Serial Number (ISSN)

  • 1047-8477

Digital Object Identifier (DOI)

  • 10.1016/j.jsb.2004.03.008

Language

  • eng