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Bead movement by single kinesin molecules studied with optical tweezers

Nature volume 348, pages 348–352 (1990)Cite this article

Abstract

KINESIN, a mechanoenzyme that couples ATP hydrolysis to movement along microtubules, is thought to power vesicle transport and other forms of microtubule-based motility1–6. Here, microscopic silica beads7 were precoated with carrier protein8,9, exposed to low concentrations of kinesin, and individually manipulated with a single-beam gradient-force optical particle trap10–12 ('optical tweezers') directly onto microtubules. Optical tweezers greatly improved the efficiency of the bead assay, particularly at the lowest kinesin concentrations (corresponding to ∼1 molecule per bead). Beads incubated with excess kinesin moved smoothly along a microtubule for many micrometres, but beads carrying from 0.17–3 kinesin molecules per bead, moved, on average, only about 1.4 µm and then spontaneously released from the microtuble. Application of the optical trap directly behind such moving beads often pulled them off the microtubule and back into the centre of the trap. This did not occur when a bead was bound by an AMP.PNP-induced rigor linkage, or when beads were propelled by several kinesin molecules. Our results are consistent with a model in which kinesin detaches briefly from the microtubule during a part of each mechanochemical cycle, rather than a model in which kinesin remains bound at all times.

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Author notes

  1. Bruce J. Schnapp: To whom correspondence should be addressed.

Authors and Affiliations

  1. Rowland Institute for Science, 100 Cambridge Parkway, Cambridge, Massachusetts, 02142, USA

    Steven M. Block

  2. Department of Cellular and Development Biology, Harvard University, 16 Divinity Avenue, Cambridge, Massachusetts, 02138, USA

    Steven M. Block & Lawrence S. B. Goldstein

  3. Department of Physiology, Boston University School of Medicine, 80 East Concord Street, Boston, Massachusetts, 02192, USA

    Bruce J. Schnapp

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  1. Steven M. Block
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  2. Lawrence S. B. Goldstein
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  3. Bruce J. Schnapp
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Block, S., Goldstein, L. & Schnapp, B. Bead movement by single kinesin molecules studied with optical tweezers. Nature 348, 348–352 (1990). https://doi.org/10.1038/348348a0

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