Kinesin has three nucleotide-dependent conformations: Implications for strain-dependent release

Jun Xing, Willy Wriggers, Geraldine M. Jefferson, Richard Stein, Herbert C. Cheung, Steven Rosenfeld

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Although crystallographic information is available on several nucleotide-induced states in myosin, little is known about the corresponding structural changes in kinesin, since a crystallographic model is only available for the kinesin:ADP complex. This makes it difficult to characterize at a molecular level the structural changes that occur in this motor through the course of its ATPase cycle. In this study, we report on the production of a series of single tryptophan mutants of a monomeric human kinesin motor domain, which demonstrate nucleotide-dependent changes in microtubule affinity that are similar to wild type. We have used these mutations to measure intramolecular distances in both strong and weak binding states, using florescence resonance energy transfer. This work provides direct evidence that movement of the switch II loop and helix are essential to mediate communication between the catalytic and microtubule binding sites, evidence that is supported as well by molecular modeling. Kinetic studies of fluorescent nucleotide binding to these mutants are consistent with these distance changes, and demonstrate as well that binding of ADP produces two structural transitions, neither of which are identical to that produced by the binding of ATP. This study provides a basis for understanding current structural models of the kinesin mechanochemical cycle.

Original languageEnglish (US)
Pages (from-to)35413-35423
Number of pages11
JournalJournal of Biological Chemistry
Volume275
Issue number45
DOIs
StatePublished - Nov 10 2000
Externally publishedYes

Fingerprint

Kinesin
Conformations
Nucleotides
Microtubules
Adenosine Diphosphate
Molecular modeling
Structural Models
Energy Transfer
Myosins
Tryptophan
Energy transfer
Adenosine Triphosphatases
Adenosine Triphosphate
Binding Sites
Communication
Switches
Mutation
Kinetics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Kinesin has three nucleotide-dependent conformations : Implications for strain-dependent release. / Xing, Jun; Wriggers, Willy; Jefferson, Geraldine M.; Stein, Richard; Cheung, Herbert C.; Rosenfeld, Steven.

In: Journal of Biological Chemistry, Vol. 275, No. 45, 10.11.2000, p. 35413-35423.

Research output: Contribution to journalArticle

Xing, Jun ; Wriggers, Willy ; Jefferson, Geraldine M. ; Stein, Richard ; Cheung, Herbert C. ; Rosenfeld, Steven. / Kinesin has three nucleotide-dependent conformations : Implications for strain-dependent release. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 45. pp. 35413-35423.
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