Three-dimensional reconstruction of an actin bundle.
We present the three-dimensional structure of an actin filament bundle from the sperm of Limulus. The bundle is a motile structure which by changing its twist, converts from a coiled to an extended form. The bundle is composed of actin plus two auxiliary proteins of molecular masses 50 and 60 kD. Fraying the bundle with potassium thiocyanate created three classes of filaments: actin, actin plus the 60-kD protein, and actin plus both the auxiliary proteins. We examined these filaments by transmission electron microscopy and scanning transmission electron microscopy (STEM). Three-dimensional reconstructions from electron micrographs allowed us to visualize the actin subunit and the 60- and 50-kD subunits bound to it. The actin subunit appears to be bilobed with dimensions 70 X 40 X 35 A. The inner lobe of the actin subunit, located at 20 A radius, is a prolate ellipsoid, 50 X 25 A; the outer actin lobe, at 30 A radius, is a 35-A-diam spheroid. Attached to the inner lobe of actin is the 60-kD protein, an oblate spheroid, 55 X 40 A, at 50 A radius. The armlike 50-kD protein, at 55 A radius, links the 60-kD protein on one of actin's twin strands to the outer lobe of the actin subunit on the opposite strand. We speculate that the 60-kD protein may be a bundling protein and that the 50-kD protein may be responsible for the change in twist of the filaments which causes extension of the bundle.
Bullitt
E S
ES
Graduate Program in Biophysics, Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts 02254.
DeRosier
D J
DJ
Coluccio
L M
LM
Tilney
L G
LG
D008841:Actin Cytoskeleton / Q000032:analysis / Q000648:ultrastructure*; D000199:Actins / Q000032:analysis*; D000818:Animals; D003599:Cytoskeleton / Q000648:ultrastructure*; D004591:Electrophoresis, Polyacrylamide Gel; D005583:Fourier Analysis; D005612:Freeze Drying; D006737:Horseshoe Crabs; D007091:Image Processing, Computer-Assisted; D008297:Male; D008840:Microfilament Proteins / Q000032:analysis*; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D013094:Spermatozoa / Q000032:analysis / Q000648:ultrastructure
D000199:Actins; D008840:Microfilament Proteins