Single kinesin molecules crossbridge microtubules in vitro

S. B. Andrews, P. E. Gallant, R. D. Leapman, B. J. Schnapp, T. S. Reese

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

Kinesin is a cytoplasmic motor protein that moves along microtubules and can induce microtubule bundling and sliding in vitro. To determine how kinesin mediates microtubule interactions, we determined the shapes and mass distributions of squid brain kinesin, taxol-stabilized microtubules (squid and bovine), and adenosine 5′-[β,γ-imido]triphosphate-stabilized kinesin-microtubule complexes by high-resolution metal replication and by low-temperature, low-dose dark-field scanning transmission electron microscopy of unfixed, directly frozen preparations. Mass mapping by electron microscopy revealed kinesins loosely attached to the carbon support as asymmetrical dumbbell-shaped molecules, 40-52 nm long, with a mass of 379 ± 15 kDa. The mass distribution and shape of these molecules suggest that these images represent kinesin in a shortened conformation. Kinesin-microtubule complexes were organized as bundles of linearly arrayed microtubules, stitched together at irregular intervals by crossbridges typically ≤25 nm long. The crossbridges had a mass of 360 ±15 kDa, consistent with one kinesin per crossbridge. These results suggest that kinesin has a second microtubule binding site in addition to the known site on the motor domain of the heavy chain; this second site may be located near the C terminus of the heavy chains or on the associated light chains. Thus, kinesin could play a role in either crosslinking or sliding microtubules.

Original languageEnglish (US)
Pages (from-to)6503-6507
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number14
DOIs
StatePublished - Jul 15 1993

Keywords

  • Cytoskeleton
  • Molecular structure
  • Scanning transmission electron microscopy

ASJC Scopus subject areas

  • General

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