Structural relationships of actin, myosin, and tropomyosin revealed by cryo-electron microscopy.
We have calculated three-dimensional maps from images of myosin subfragment-1 (S1)-decorated thin filaments and S1-decorated actin filaments preserved in frozen solution. By averaging many data sets we obtained highly reproducible maps that can be interpreted simply to provide a model for the native structure of decorated filaments. From our results we have made the following conclusions. The bulk of the actin monomer is approximately 65 X 40 X 40 A and is composed of two domains. In the filaments the monomers are strongly connected along the genetic helix with weaker connections following the long pitch helix. The long axis of the monomer lies roughly perpendicular to the filament axis. The myosin head (S1) approaches the actin filament tangentially and binds to a single actin, the major interaction being with the outermost domain of actin. In the map the longest chord of S1 is approximately 130 A. The region of S1 closest to actin is of high density, whereas the part furthest away is poorly defined and may be disordered. By comparing maps from decorated thin filaments with those from decorated actin, we demonstrate that tropomyosin is bound to the inner domain of actin just in front of the myosin binding site at a radius of approximately 40 A. A small change in the azimuthal position of tropomyosin, as has been suggested by others to occur during Ca2+-mediated regulation in vertebrate striated muscle, appears to be insufficient to eclipse totally the major site of interaction between actin and myosin.
D008841:Actin Cytoskeleton / Q000032:analysis / Q000648:ultrastructure*; D000199:Actins / Q000032:analysis*; D000818:Animals; D003599:Cytoskeleton / Q000648:ultrastructure*; D005615:Freezing; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron / Q000379:methods; D009119:Muscle Contraction; D009218:Myosins / Q000032:analysis*; D011817:Rabbits; D014335:Tropomyosin / Q000032:analysis*
D000199:Actins; D014335:Tropomyosin; D009218:Myosins