Three-dimensional structure of frozen-hydrated paracrystals of myosin rod.
Myosin rod, the tail fragment of myosin, aggregates into various structures at physiological ionic strength. One form, the type II paracrystal, a thin, ribbon-like species, has recently been studied using conventional electron microscopy and was shown to possess two-dimensional order and, further, is likely to be useful in the investigation of the arrangement of myosin molecules in the backbone of the vertebrate muscle thick filament (Ward & Bennett, 1989). We have now examined this aggregate in the frozen-hydrated state by cryo-electron microscopy. Image analysis indicated that the projected structure has the same p12, plane group symmetry as seen after negative staining. A three-dimensional map, calculated from projections, indicated that the structure comprises a bilayer arrangement of strands of grouped rod molecules, with the strands parallel in each layer. The layers themselves are related by screw symmetry. Strands in adjacent layers have opposing polarity with their long axes at an angle of 32 degree to each other. Protein density measurements, carried out on unstained specimens using electron energy-loss spectroscopy, showed that the 44 X 13 X 13 nm unit cell is composed of 40% protein. The density measurements indicated that 9-12 rod molecules pass through each strand. Modelling rod molecules with 43 nm parallel overlaps in a body-centered tetragonal lattice produced a strand that compared favorably with the reconstructed strand. The size and content of these strands suggests that they are analogous to subfilaments observed in the vertebrate myosin thick filament.
Ward
R
R
Department of Anatomy, University of Pennsylvania, Philadelphia 19104.
Murray
J M
JM
D000818:Animals; D003460:Crystallization; D005615:Freezing; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron; D015394:Molecular Structure; D009218:Myosins / Q000737:chemistry / Q000648:ultrastructure*; D011487:Protein Conformation; D011817:Rabbits
D009218:Myosins