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High-Resolution Structural Analysis of the Kinesin-Microtubule Complex by Electron Cryo-Microscopy

  • Keiko Hirose
  • Linda A. Amos
Part of the Methods in Molecular Biology™ book series (MIMB, volume 392)

Abstract

To understand the interaction of kinesin and microtubules, it is necessary to study the three-dimensional (3D) structures of the kinesin-microtubule complex at a high enough resolution to identify structural components such as α-helices and β-sheets. Electron cryo-microscopy combined with computer image analysis is the most common method to study such complexes that cannot be crystallized. By selecting microtubules that have a helical symmetry, 3D structures of the complex can be calculated using the helical 3D reconstruction method. Details of the interaction are studied by docking the individual crystal structures of the kinesin motor domains and tubulin heterodimer into the 3D maps of the complex. To study the structural changes during ATP hydrolysis, structures of the complexes in the presence and absence of different nucleotides are compared.

Key Words

Kinesin microtubule molecular motors electron microscopy three-dimensional reconstruction 

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Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Keiko Hirose
    • 1
  • Linda A. Amos
    • 2
  1. 1.Gene Function Research CenterNational Institute of Advanced Industrial Science and Technology (AIST)TsukubaJapan
  2. 2.MRC Laboratory of Molecular BiologyCambridgeUK

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