Structure of myosin/paramyosin filaments from a molluscan smooth muscle

Abstract

Small-angle X-ray diffraction patterns of chemically skinned pedal retractor muscles from Mytilus (PRM) in the relaxed state show a set of diffuse off-meridional reflections that arise from a helical array of myosin crossbridges with 8/3 screw symmetry. Experiments involving extraction of myosin as well as analysis of the rigor pattern have been used to confirm the origin of these reflections. The relatively high myosin/paramyosin molar ratio (1.3 to 1.6) in PRM compared to other molluscan smooth muscles may account for the observation of the relatively stronger diffraction from the myosin array. Thick filaments isolated from PRM and contrasted by negative staining for electron microscopy appear to be very long (up to 17 micron), and to have a rather small diameter (about 40 nm at the center); they taper gradually toward the ends. These filaments show a clear transverse band pattern repeating at 14.4 nm and elongated projections (crossbridges) at the surface except in the central bare zone. Optical diffraction patterns show reflections from crossbridges consistent with the X-ray patterns of the relaxed whole muscle. Filaments unidirectionally shadowed with platinum show diagonal striations running at an angle of about 17 degrees to the filament axis, revealing that the crossbridges are arrayed in a right-handed helix. The paramyosin core is clearly seen upon extraction of myosin. Observations on both negatively stained and sectioned material are consistent with the results of Elliott (1979) and Bennett & Elliott (1981) suggesting a layered structure of the core. Cores stripped of myosin, however, appear to undergo some distortion indicating that the three-dimensional structure is not yet completely solved. The assembly of these thick filaments presents intriguing structural problems since the myosin surface lattice does not appear to have the same symmetry as the underlying paramyosin core.