Snapshots of Kinesin Motors on Microtubule Tracks
Kinesin motors couple ATP hydrolysis to movement along microtubules, which act both as tracks and as activators of kinesin ATPase activity. Cryo-electron microscopy and image processing enables generation of three-dimensional snapshots of kinesin motors on their tracks at different stages of their ATPase cycle, and can reveal their motor mechanisms at secondary structure resolution. Here, we describe in detail the methods and conditions employed in our lab to prepare high-quality frozen-hydrated samples, which yield structural insights into kinesin motor mechanisms.
Key wordsKinesin Microtubules Co-sedimentation assay Negative stain electron microscopy Cryo-electron microscopy
The authors would like to thank past and present members of the Moores Lab (Andy Bodey, Carsten Peters, Christina Hoey, and Kanwal Zehra) for helpful discussions. We are grateful to Kanwal Zehra and Natasha Lukoyanova for providing us with constructive comments on the manuscript. We thank the Wellcome Trust, the Birth Defects Foundation, the Ecole Normale Supérieure, and the French Ministère de la Recherche for funding.