Kinetic overshoot in actin network assembly induced jointly by branching and capping proteins

We report an experimental study of the kinetics of actin assembly mediated by branching and capping proteins. Our findings confirm the recent prediction of a "branching explosion" occurring during polymerization. Fluorescence imaging shows a number of actin filaments with branches within a few minutes of polymerization, induced by the activated branching protein complex Arp2/3, but the number of visible branches decreases over time. The light-scattering intensity displays an overshoot as a function of time, which we attribute to the formation of highly branched clusters early in polymerization. Furthermore, the overshoot occurs over a limited range of the ratio of concentrations of branching and capping proteins, also consistent with the theoretical model. These results establish a natural link between the kinetic theory of actin assembly in vitro and the cytoskeletal structure and actin dynamics in motile cells.