title: Kinetics of actin monomer exchange at the slow growing ends of actin filaments and their relation to the elongation of filaments shortened by gelsolin.
abstract: The kinetics of actin monomer exchange with the slow growing pointed ends of actin filaments have been determined by measuring rates of monomer addition to or loss from filaments with their fast-growing barbed ends blocked by the protein gelsolin. Direct measurements of filament length by electron microscopy confirmed that each gelsolin acts as a nucleus for an actin filament. The rate constants ascertained are k- = 0.03 s-1; k+ = 0.06 microM-1 s-1 at 23 degrees C and k- = 0.11 s-1; k+ = 0.09 microM-1 s-1 at 37 degrees C. They are approximately independent of pH from 7.0 to 8.0 at both temperatures. These rates are far slower than those reported on the basis of some electron microscopic studies of filaments assembled on to actin bundles. The rate constants also predict a higher critical monomer concentration for the pointed end at 37 degrees C than at room temperature, consistent with direct measurements of this quantity. The relative slowness of the monomer exchange at the pointed end suggests that actin filaments with blocked barbed ends are relatively stable. The rate of redistribution of actin monomers from filaments stabilized at their barbed ends by gelsolin-calcium complex to longer filaments was measured following removal of Ca2+, which decreases the capacity of gelsolin to nucleate filaments. The elongation occurs at a rate consistent with the measured rates of monomer exchange and is quantitatively described by Hill's model for filament elongation by random exchange of monomers from one end.
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D002135:Calcium-Binding Proteins / Q000494:pharmacology; D018260:Gelsolin; D066298:In Vitro Techniques; D007700:Kinetics; D008840:Microfilament Proteins / Q000494:pharmacology; D008854:Microscopy, Electron; D009132:Muscles / Q000378:metabolism; D011487:Protein Conformation / Q000187:drug effects; D011817:Rabbits
chemical_list: D000199:Actins; D002135:Calcium-Binding Proteins; D018260:Gelsolin; D008840:Microfilament Proteins