Abstract
Single actin filaments (∼7nm in diameter) labelled with fluorescent phalloidin can be clearly seen by video-fluorescence microscopy1. This technique has been used to observe motions of single filaments in solution and in several in vitro movement assays1–5. In a further development of the technique, we report here a method to catch and manipulate a single actin filament (F-actin) by glass microneedles under conditions in which external force on the filament can be applied and measured. Using this method, we directly measured the tensile strength of a filament (the force necessary to break the bond between two actin monomers) and the force required for a filament to be moved by myosin or its proteolytic fragment bound to a glass surface in the presence of ATP. The first result shows that the tensile strength of the F-actin–phalloidin complex is comparable with the average force exerted on a single thin filament in muscle fibres during isometric contraction. This force is increased only slightly by tropomyosin. The second measurement shows that the myosin head (subfragment-1) can produce the same ATP-dependent force as intact myosin. The magnitude of this force is comparable with that produced by each head of myosin in muscle during isometric contraction.
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Kishino, A., Yanagida, T. Force measurements by micromanipulation of a single actin filament by glass needles. Nature 334, 74–76 (1988). https://doi.org/10.1038/334074a0
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DOI: https://doi.org/10.1038/334074a0
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