Pathway of ADP-stimulated ADP release and dissociation of tethered kinesin from microtubules. Implications for the extent of processivity

Biochemistry
D D Hackney

Abstract

Kinesin binds to microtubules with half-site ADP release to form a tethered intermediate with one attached head without nucleotide and one tethered head that retains its bound ADP. For DKH405 containing amino acid residues 1-405 of Drosophila kinesin, release of the remaining ADP from the tethered head is slow (0.05 s(-1)), but release is accelerated by added ADP or ATP. The maximum rate of ADP-stimulated dissociation of tethered DKH405 from the microtubule is approximately 12 s(-1) as determined by turbidity. Parallel measurements of ADP-stimulated release of 2'(3')-O-(N-methylanthraniloyl)-ADP (mantADP) from the tethered intermediate by fluorescence indicate that the reaction is biphasic with a fast phase that occurs at a rate that is similar to dissociation. The rate of the slow phase is dependent on the concentrations of salt and microtubules and is equal in each case to the rate for bimolecular stimulation of ADP release by microtubules as measured independently. These results are consistent with a scheme in which the fast phase, with approximately one-third of the total amplitude change, is due to ADP-stimulated release of mantADP from the tethered intermediate at approximately 6 s(-1). This direct release of mantADP cont...Continue Reading

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Citations

Mar 10, 2004·Biochemistry·Lisa M KlumppSusan P Gilbert
Oct 7, 2009·Proceedings of the National Academy of Sciences of the United States of America·Sander VerbruggeErwin J G Peterman
Mar 11, 2003·The Journal of Biological Chemistry·Steven S RosenfeldSteven M Block
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Nov 17, 2010·PLoS Computational Biology·Matthew L KutysWilliam O Hancock
Dec 13, 2005·Proceedings of the National Academy of Sciences of the United States of America·David D Hackney
Feb 27, 2004·Proceedings of the National Academy of Sciences of the United States of America·Lisa M KlumppSusan P Gilbert
Jan 22, 2004·Proceedings of the National Academy of Sciences of the United States of America·William R SchiefJonathon Howard
Dec 3, 2013·Bioinformatics·Hamidreza Khataee, Alan Wee-Chung Liew
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