A model for kinesin movement from nanometer-level movements of kinesin and cytoplasmic dynein and force measurements.
Our detailed measurements of the movements of kinesin- and dynein-coated latex beads have revealed several important features of the motors which underlie basic mechanical aspects of the mechanisms of motor movements. Kinesin-coated beads will move along the paths of individual microtubule protofilaments with high fidelity and will pause at 4 nm intervals along the microtubule axis under low ATP conditions. In contrast, cytoplasmic dynein-coated beads move laterally across many protofilaments as they travel along the microtubule, without any regular pauses, suggesting that the movements of kinesin-coated beads are not an artefact of the method. These kinesin bead movements suggest a model for kinesin movement in which the two heads walk along an individual protofilament in a hand-over-hand fashion. A free head would only be able to bind to the next forward tubulin subunit on the protofilament and its binding would pull off the trailing head to start the cycle again. This model is consistent with the observed cooperativity between the heads and with the movement by single dimeric molecules. Several testable predictions of the model are that kinesin should be able to bind to both alpha and beta tubulin and that the length of the neck region of the molecule should control the off-axis motility. In this article, we describe the technology for measuring nanometer-level movements and the force generated by the kinesin molecule.
Kuo
S C
SC
Department of Cell Biology, Duke University Medical Center, Durham, NC 27710.
Gelles
J
J
Steuer
E
E
Sheetz
M P
MP
D000251:Adenosine Triphosphatases / Q000502:physiology*; D002138:Calibration; D003593:Cytoplasm / Q000502:physiology; D004398:Dyneins / Q000502:physiology*; D004581:Electronics; D007091:Image Processing, Computer-Assisted; D016547:Kinesins; D007700:Kinetics; D008954:Models, Biological; D055095:Optics and Photonics; D010784:Photomicrography; D014743:Videotape Recording
D000251:Adenosine Triphosphatases; D004398:Dyneins; D016547:Kinesins