Electron microscopic determination of the actin filament end at which cytochalasin B blocks monomer addition using the acrosomal actin bundle from horseshoe crab sperm.
G-actin freed from exogenous ATP was added to the pieces of isolated acrosomal actin bundles from horseshoe crab sperm to form filaments as reported earlier (Tilney, L.G., Bonder, E.M., & DeRosier, D.J. (1981) J. Cell Biol. 90, 485-494). The growth of a filament was far more rapid at one end (the preferred end) than the other end. These ends were shown to correspond to the barbed and pointed ends, respectively, by decoration of the filaments with myosin subfragment 1. Cytochalasin B inhibited the monomer addition at the preferred end. This technique is useful in determining the ends to which actin filament end-binding proteins from nonmuscle cells bind, which are considered to regulate the actin polymerization in the cells.
D000177:Acrosome / Q000378:metabolism / Q000648:ultrastructure*; D000199:Actins / Q000378:metabolism*; D000818:Animals; D003571:Cytochalasin B / Q000494:pharmacology*; D003599:Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure*; D006737:Horseshoe Crabs; D007700:Kinetics; D046911:Macromolecular Substances; D008297:Male; D008854:Microscopy, Electron; D013094:Spermatozoa / Q000648:ultrastructure*
D000199:Actins; D046911:Macromolecular Substances; D003571:Cytochalasin B