Direct measurement of force generation by actin filament polymerization using an optical trap

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Abstract
Actin filament polymerization generates force for protrusion of the leading edge in motile cells. In protrusive structures, multiple actin filaments are arranged in cross-linked webs (as in lamellipodia or pseudopodia) or parallel bundles (as in filopodia). We have used an optical trap to directly measure the forces generated by elongation of a few parallel-growing actin filaments brought into apposition with a rigid barrier, mimicking the geometry of filopodial protrusion. We find that the growth of approximately eight actin parallel-growing filaments can be stalled by relatively small applied load forces on the order of 1 pN, consistent with the theoretical load required to stall the elongation of a single filament under our conditions. Indeed, large length fluctuations during the stall phase indicate that only the longest actin filament in the bundle is in contact with the barrier at any given time. These results suggest that force generation by small actin bundles is limited by a dynamic instability of single actin filaments, and therefore living cells must use actin-associated factors to suppress this instability to generate substantial forces by elongation of parallel bundles of actin filaments.