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Microscopic evidence for a minus-end-dir ... stroke in the kinesin motor ncd
Microscopic evidence for a minus-end-directed power stroke in the kinesin motor ncd.
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We used cryo-electron microscopy and image reconstruction to investigate the structure and microtubule-binding configurations of dimeric non-claret disjunctional (ncd) motor domains under various nucleotide conditions, and applied molecular docking using ncd's dimeric X-ray structure to generate a mechanistic model for force transduction. To visualize the alpha-helical coiled-coil neck better, we engineered an SH3 domain to the N-terminal end of our ncd construct (296-700). Ncd exhibits strikingly different nucleotide-dependent three-dimensional conformations and microtubule-binding patterns from those of conventional kinesin. In the absence of nucleotide, the neck adapts a configuration close to that found in the X-ray structure with stable interactions between the neck and motor core domain. Minus-end-directed movement is based mainly on two key events: (i) the stable neck-core interactions in ncd generate a binding geometry between motor and microtubule which places the motor ahead of its cargo in the minus-end direction; and (ii) after the uptake of ATP, the two heads rearrange their position relative to each other in a way that promotes a swing of the neck in the minus-end direction.
Wendt TG, Volkmann N, Skiniotis G, Goldie KN, Müller J, Mandelkow E, Hoenger A
The EMBO journal
2002-11-15 00:00
21
22
5969-78
Adenosine Diphosphate,Adenosine Triphosphate,Animals,Cryoelectron Microscopy,Drosophila Proteins,Image Processing, Computer-Assisted,Kinesin,Microtubules,Mitotic Spindle Apparatus,Models, Molecular,Molecular Motor Proteins,Movement,Protein Conformation,Protein Structure, Tertiary,Recombinant Fusion Proteins,Structure-Activity Relationship,Tubulin,Drosophila Proteins,Molecular Motor Proteins,Recombinant Fusion Proteins,Tubulin,ncd protein, Drosophila,Adenosine Triphosphate,Adenosine Diphosphate,Kinesin
European Molecular Biology Laboratory, Structure Programme, Meyerhofstrasse 1, D-69117 Heidelberg, Germany
EMBO J.
NIGMS GM 64473
0261-4189
561
True
12426369
