Dissociation of double-headed cytoplasmic dynein into single-headed species and its motile properties

Cell Motility and the Cytoskeleton
Shiori Toba, Yoko Y Toyoshima

Abstract

Cytoplasmic dynein is a minus-end directed microtubule motor and plays important roles in the transport of various intracellular cargoes. Cytoplasmic dynein comprises two identical heavy chains and forms a dimer (double-headed dynein); the total molecular weight of the cytoplasmic dynein complex is about 1.5 million. The dynein motor domain is structurally very different from those of kinesin and myosin, and our understanding of the mechanisms of dynein energy transduction is limited mainly because of the difficulty in obtaining a sufficient quantity of purified and active cytoplasmic dynein. We purified cytoplasmic dynein, which was free from dynactin and other dynein-associated proteins. The purified cytoplasmic dynein was active in an in vitro motility assay. The controlled dialysis of the purified dynein against 4 M urea resulted in its complete dissociation into monomeric species (single-headed dynein). The separation of the dynein heads by the treatment was reversible. The MgATPase activities of the single-headed and reconstituted double-headed dynein were comparable to that of intact dynein. The double-headed dynein bundled microtubules in the absence of ATP; the single-headed dynein did not. The single-headed dynein pro...Continue Reading

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Citations

Aug 25, 2004·Proceedings of the National Academy of Sciences of the United States of America·Yoshinori TakahashiYoko Y Toyoshima
Mar 14, 2012·Proceedings of the National Academy of Sciences of the United States of America·Wilhelm J WalterWalter Steffen
Nov 3, 2010·The Journal of Biological Chemistry·Takayuki TorisawaYoko Y Toyoshima
Dec 26, 2008·Molecular Biology of the Cell·Jessica L CrowleyElizabeth J Luna
Dec 15, 2010·Molecular Biology of the Cell·Shiori TobaWinfield S Sale
Apr 6, 2006·Proceedings of the National Academy of Sciences of the United States of America·Shiori TobaHideo Higuchi
Aug 8, 2009·Journal of Neurochemistry·Nariko ArimuraKozo Kaibuchi
Nov 15, 2005·Biophysical Journal·Yi Qin Gao
Sep 12, 2008·The EMBO Journal·Masami YamadaShinji Hirotsune
Dec 8, 2009·Journal of Structural Biology·Wilhelm J WalterWalter Steffen
Nov 24, 2004·Current Biology : CB·Roop Mallik, Steven P Gross
Feb 7, 2017·Biology·Tarun M Kapoor
Dec 24, 2018·FEBS Letters·Vincent A BehrensWalter Steffen

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