2006年12月
Identification of a strong binding site for kinesin on the microtubule using mutant analysis of tubulin
EMBO JOURNAL
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- 巻
- 25
- 号
- 24
- 開始ページ
- 5932
- 終了ページ
- 5941
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1038/sj.emboj.7601442
- 出版者・発行元
- NATURE PUBLISHING GROUP
The kinesin-binding site on the microtubule has not been identified because of the technical difficulties involved in the mutant analyses of tubulin. Exploiting the budding yeast expression system, we succeeded in replacing the negatively charged residues in the alpha-helix 12 of beta-tubulin with alanine and analyzed their effect on kinesin-microtubule interaction in vitro. The microtubule gliding assay showed that the affinity of the microtubules for kinesin was significantly reduced in E410A, D417A, and E421A, but not in E412A mutant. The unbinding force measurement revealed that in the former three mutants, the kinesin-microtubule interaction in the adenosine 5'-[beta,gamma imido] triphosphate state (AMP-PNP state) became less stable when a load was imposed towards the microtubule minus end. In parallel with this decreased stability, the stall force of kinesin was reduced. Our results implicate residues E410, D417, and E421 as crucial for the kinesin-microtubule interaction in the strong binding state, thereby governing the size of kinesin stall force.
- リンク情報
- ID情報
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- DOI : 10.1038/sj.emboj.7601442
- ISSN : 0261-4189
- Web of Science ID : WOS:000242891100028