Structural studies of kinesin-nucleotide intermediates.

The Journal of Biological Chemistry
S S RosenfeldH C Cheung

Abstract

We have investigated the structural changes that occur in the molecular motor kinesin during its ATPase cycle, utilizing two bacterially expressed constructs. The structure of both constructs has been examined as a function of the nature of the nucleotide intermediate occupying the active site by means of sedimentation velocity, sedimentation equilibrium, fluorescence solute quenching, fluorescence anisotropy decay, and limited proteolysis. While the molecular weight of monomeric and dimeric human kinesin constructs, as measured by sedimentation velocity and sedimentation equilibrium, and the tryptic cleavage pattern are unaffected by the nucleotide intermediate occupying the active site, significant changes in the rotational correlation time of fluorescently labeled kinesin-nucleotide intermediates can be detected. These results suggest that kinesin contains an internal "hinge" whose flexibility varies through the course of the ATPase cycle. In prehydrolytic, "strong" binding states, this hinge is relatively rigid, while in posthydrolytic, "weak" binding states, it is more flexible. Our results, in conjunction with anisotropy decay studies of myosin, suggest that these two molecular motors may share a common structural feature...Continue Reading

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Citations

Mar 8, 2011·Journal of Molecular Biology·P Keith VeroneseAaron L Lucius
Oct 6, 2000·The Journal of Biological Chemistry·F KozielskiM H Koch
Jul 21, 2005·The Journal of Biological Chemistry·Peter HöökRichard B Vallee
Apr 16, 1998·The Journal of Biological Chemistry·H L SweeneyJ R Sellers
Oct 24, 1998·The Journal of Biological Chemistry·S S RosenfeldH L Sweeney
Jul 18, 2002·The Journal of Biological Chemistry·Steven S RosenfeldPeter H King
Jan 26, 2006·The Journal of Biological Chemistry·Zoltan MaligaSteven S Rosenfeld
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Jun 15, 2000·The Journal of Biological Chemistry·J XingS S Rosenfeld
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