Bead movement by single kinesin molecules studied with optical tweezers

Steven M. Block, Lawrence S B Goldsteint, Bruce Schnapp

Research output: Contribution to journalArticle

766 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)348-352
Number of pages5
JournalNature
Volume348
Issue number6299
StatePublished - Nov 22 1990
Externally publishedYes

Fingerprint

Optical Tweezers
Kinesin
Microtubules
Transport Vesicles
Adenosine Monophosphate
Silicon Dioxide
Hydrolysis
Adenosine Triphosphate

ASJC Scopus subject areas

  • General

Cite this

Block, S. M., Goldsteint, L. S. B., & Schnapp, B. (1990). Bead movement by single kinesin molecules studied with optical tweezers. Nature, 348(6299), 348-352.

Bead movement by single kinesin molecules studied with optical tweezers. / Block, Steven M.; Goldsteint, Lawrence S B; Schnapp, Bruce.

In: Nature, Vol. 348, No. 6299, 22.11.1990, p. 348-352.

Research output: Contribution to journalArticle

Block, SM, Goldsteint, LSB & Schnapp, B 1990, 'Bead movement by single kinesin molecules studied with optical tweezers', Nature, vol. 348, no. 6299, pp. 348-352.
Block SM, Goldsteint LSB, Schnapp B. Bead movement by single kinesin molecules studied with optical tweezers. Nature. 1990 Nov 22;348(6299):348-352.
Block, Steven M. ; Goldsteint, Lawrence S B ; Schnapp, Bruce. / Bead movement by single kinesin molecules studied with optical tweezers. In: Nature. 1990 ; Vol. 348, No. 6299. pp. 348-352.
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