title: Head-head interactions of resting myosin crossbridges in intact frog skeletal muscles, revealed by synchrotron x-ray fiber diffraction.
abstract: The intensities of the myosin-based layer lines in the x-ray diffraction patterns from live resting frog skeletal muscles with full thick-thin filament overlap from which partial lattice sampling effects had been removed were analyzed to elucidate the configurations of myosin crossbridges around the thick filament backbone to nanometer resolution. The repeat of myosin binding protein C (C-protein) molecules on the thick filaments was determined to be 45.33 nm, slightly longer than that of myosin crossbridges. With the inclusion of structural information for C-proteins and a pre-powerstroke head shape, modeling in terms of a mixed population of regular and perturbed regions of myosin crown repeats along the filament revealed that the myosin filament had azimuthal perturbations of crossbridges in addition to axial perturbations in the perturbed region, producing pseudo-six-fold rotational symmetry in the structure projected down the filament axis. Myosin crossbridges had a different organization about the filament axis in each of the regular and perturbed regions. In the regular region that lacks C-proteins, there were inter-molecular interactions between the myosin heads in axially adjacent crown levels. In the perturbed region that contains C-proteins, in addition to inter-molecular interactions between the myosin heads in the closest adjacent crown levels, there were also intra-molecular interactions between the paired heads on the same crown level. Common features of the interactions in both regions were interactions between a portion of the 50-kDa-domain and part of the converter domain of the myosin heads, similar to those found in the phosphorylation-regulated invertebrate myosin. These interactions are primarily electrostatic and the converter domain is responsible for the head-head interactions. Thus multiple head-head interactions of myosin crossbridges also characterize the switched-off state and have an important role in the regulation or other functions of myosin in thin filament-regulated muscles as well as in the thick filament-regulated muscles.
authors.affiliation: Institute for Protein Research, Osaka University, Suita, Osaka, Japan.
authors.forename: Kanji
authors.initials: K
authors.lastname: Oshima
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Sugimoto
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Irving
authors.forename: Katsuzo
authors.initials: K
authors.lastname: Wakabayashi
mesh_terms: D008841:Actin Cytoskeleton; D000818:Animals; D066298:In Vitro Techniques; D008954:Models, Biological; D008958:Models, Molecular; D018482:Muscle, Skeletal / Q000378:metabolism*; D009218:Myosins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011898:Ranidae; D012146:Rest / Q000502:physiology*; D040681:Structural Homology, Protein; D017356:Synchrotrons; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D009218:Myosins