3D structure of relaxed fish muscle myosin filaments by single particle analysis

Hind A. AL-Khayat, Edward P. Morris, Robert W. Kensler, John M. Squire

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

To understand the structural changes involved in the force-producing myosin cross-bridge cycle in vertebrate muscle it is necessary to know the arrangement and conformation of the myosin heads at the start of the cycle (i.e. the relaxed state). Myosin filaments isolated from goldfish muscle under relaxing conditions and viewed in negative stain by electron microscopy (EM) were divided into segments and subjected to three-dimensional (3D) single particle analysis without imposing helical symmetry. This allowed the known systematic departure from helicity characteristic of vertebrate striated muscle myosin filaments to be preserved and visualised. The resulting 3D reconstruction reveals details to about 55 Å resolution of the myosin head density distribution in the three non-equivalent head 'crowns' in the 429 Å myosin filament repeat. The analysis maintained the well-documented axial perturbations of the myosin head crowns and revealed substantial azimuthal perturbations between crowns with relatively little radial perturbation. Azimuthal rotations between crowns were approximately 60°, 60° and 0°, rather than the regular 40° characteristic of an unperturbed helix. The new density map correlates quite well with the head conformations analysed in other EM studies and in the relaxed fish muscle myosin filament structure modelled from X-ray fibre diffraction data. The reconstruction provides information on the polarity of the myosin head array in the A-band, important in understanding the geometry of the myosin head interaction with actin during the cross-bridge cycle, and supports a number of conclusions previously inferred by other methods. The observed azimuthal head perturbations are consistent with the X-ray modelling results from intact muscle, indicating that the observed perturbations are an intrinsic property of the myosin filaments and are not induced by the proximity of actin filaments in the muscle A-band lattice. Comparison of the axial density profile derived in this study with the axial density profile of the X-ray model of the fish myosin filaments which was restricted to contributions from the myosin heads allows the identification of a non-myosin density peak associated with the azimuthally perturbed head crown which can be interpreted as a possible location for C-protein or X-protein (MyBP-C or -X). This position for C-protein is also consistent with the C-zone interference function deduced from previous analysis of the meridional X-ray pattern from frog muscle. It appears that, along with other functions, C-(X-) protein may have the role of slewing the heads of one crown so that they do not clash with the neighbouring actin filaments, but are readily available to interact with actin when the muscle is activated.

Original languageEnglish
Pages (from-to)202-217
Number of pages16
JournalJournal of Structural Biology
Volume155
Issue number2
DOIs
Publication statusPublished - Aug 2006
Externally publishedYes

Fingerprint

Myosins
Fishes
Muscles
Crowns
Protein C
Head
X-Rays
Actin Cytoskeleton
Vertebrates
Actins
Electron Microscopy
Skates (Fish)
Goldfish
Striated Muscle
X-Ray Diffraction
Anura
Coloring Agents

Keywords

  • 3D reconstruction
  • Fish muscle
  • MyBP-C
  • Myosin filaments
  • Single particle analysis

ASJC Scopus subject areas

  • Structural Biology

Cite this

3D structure of relaxed fish muscle myosin filaments by single particle analysis. / AL-Khayat, Hind A.; Morris, Edward P.; Kensler, Robert W.; Squire, John M.

In: Journal of Structural Biology, Vol. 155, No. 2, 08.2006, p. 202-217.

Research output: Contribution to journalArticle

AL-Khayat, Hind A. ; Morris, Edward P. ; Kensler, Robert W. ; Squire, John M. / 3D structure of relaxed fish muscle myosin filaments by single particle analysis. In: Journal of Structural Biology. 2006 ; Vol. 155, No. 2. pp. 202-217.
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