Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
The Kinesin-5 Chemomechanical Cycle Is Dominated by a Two-heads-bound State.
Chen GY, Mickolajczyk KJ, Hancock WO.
???displayArticle.abstract???
Single-molecule microscopy and stopped-flow kinetics assays were carried out to understand the microtubule polymerase activity of kinesin-5 (Eg5). Four lines of evidence argue that the motor primarily resides in a two-heads-bound (2HB) state. First, upon microtubule binding, dimeric Eg5 releases both bound ADPs. Second, microtubule dissociation in saturating ADP is 20-fold slower for the dimer than for the monomer. Third, ATP-triggered mant-ADP release is 5-fold faster than the stepping rate. Fourth, ATP binding is relatively fast when the motor is locked in a 2HB state. Shortening the neck-linker does not facilitate rear-head detachment, suggesting a minimal role for rear-head-gating. This 2HB state may enable Eg5 to stabilize incoming tubulin at the growing microtubule plus-end. The finding that slowly hydrolyzable ATP analogs trigger slower nucleotide release than ATP suggests that ATP hydrolysis in the bound head precedes stepping by the tethered head, leading to a mechanochemical cycle in which processivity is determined by the race between unbinding of the bound head and attachment of the tethered head.
Andreasson,
Examining kinesin processivity within a general gating framework.
2015, Pubmed
Andreasson,
Examining kinesin processivity within a general gating framework.
2015,
Pubmed Arpağ,
Transport by populations of fast and slow kinesins uncovers novel family-dependent motor characteristics important for in vivo function.
2014,
Pubmed
,
Xenbase Avunie-Masala,
Phospho-regulation of kinesin-5 during anaphase spindle elongation.
2011,
Pubmed Behnke-Parks,
Loop L5 acts as a conformational latch in the mitotic kinesin Eg5.
2011,
Pubmed Block,
Kinesin motor mechanics: binding, stepping, tracking, gating, and limping.
2007,
Pubmed Chen,
Kinesin-5 is a microtubule polymerase.
2015,
Pubmed
,
Xenbase Chen,
Processivity of the kinesin-2 KIF3A results from rear head gating and not front head gating.
2015,
Pubmed Chen,
Molecular counting by photobleaching in protein complexes with many subunits: best practices and application to the cellulose synthesis complex.
2014,
Pubmed Cochran,
ATPase mechanism of Eg5 in the absence of microtubules: insight into microtubule activation and allosteric inhibition by monastrol.
2005,
Pubmed Cochran,
Mechanistic analysis of the mitotic kinesin Eg5.
2004,
Pubmed Cochran,
Pathway of ATP hydrolysis by monomeric kinesin Eg5.
2006,
Pubmed Fridman,
Kinesin-5 Kip1 is a bi-directional motor that stabilizes microtubules and tracks their plus-ends in vivo.
2013,
Pubmed Gardner,
Chromosome congression by Kinesin-5 motor-mediated disassembly of longer kinetochore microtubules.
2008,
Pubmed Gerson-Gurwitz,
Directionality of individual kinesin-5 Cin8 motors is modulated by loop 8, ionic strength and microtubule geometry.
2011,
Pubmed Goulet,
Comprehensive structural model of the mechanochemical cycle of a mitotic motor highlights molecular adaptations in the kinesin family.
2014,
Pubmed Goulet,
New insights into the mechanism of force generation by kinesin-5 molecular motors.
2013,
Pubmed Guydosh,
Backsteps induced by nucleotide analogs suggest the front head of kinesin is gated by strain.
2006,
Pubmed Hackney,
Kinesin ATPase: rate-limiting ADP release.
1988,
Pubmed Hackney,
The tethered motor domain of a kinesin-microtubule complex catalyzes reversible synthesis of bound ATP.
2005,
Pubmed Hackney,
Evidence for alternating head catalysis by kinesin during microtubule-stimulated ATP hydrolysis.
1994,
Pubmed Hackney,
Highly processive microtubule-stimulated ATP hydrolysis by dimeric kinesin head domains.
1995,
Pubmed Hackney,
Nucleotide-free kinesin hydrolyzes ATP with burst kinetics.
1989,
Pubmed Hariharan,
Insights into the Mechanical Properties of the Kinesin Neck Linker Domain from Sequence Analysis and Molecular Dynamics Simulations.
2009,
Pubmed Jiang,
Influence of the kinesin neck domain on dimerization and ATPase kinetics.
1997,
Pubmed Kaan,
The structure of the kinesin-1 motor-tail complex reveals the mechanism of autoinhibition.
2011,
Pubmed Kapitein,
The bipolar mitotic kinesin Eg5 moves on both microtubules that it crosslinks.
2005,
Pubmed
,
Xenbase Kapoor,
Probing spindle assembly mechanisms with monastrol, a small molecule inhibitor of the mitotic kinesin, Eg5.
2000,
Pubmed
,
Xenbase Kaseda,
Single-headed mode of kinesin-5.
2008,
Pubmed
,
Xenbase Krzysiak,
Getting in sync with dimeric Eg5. Initiation and regulation of the processive run.
2008,
Pubmed Krzysiak,
Dimeric Eg5 maintains processivity through alternating-site catalysis with rate-limiting ATP hydrolysis.
2006,
Pubmed Larson,
Design and construction of a multiwavelength, micromirror total internal reflectance fluorescence microscope.
2014,
Pubmed Lebel,
Gold rotor bead tracking for high-speed measurements of DNA twist, torque and extension.
2014,
Pubmed Ma,
Interacting head mechanism of microtubule-kinesin ATPase.
1997,
Pubmed McCoy,
Physical limits on kinesin-5-mediated chromosome congression in the smallest mitotic spindles.
2015,
Pubmed Mickolajczyk,
Kinetics of nucleotide-dependent structural transitions in the kinesin-1 hydrolysis cycle.
2015,
Pubmed Milic,
Kinesin processivity is gated by phosphate release.
2014,
Pubmed Muretta,
Loop L5 assumes three distinct orientations during the ATPase cycle of the mitotic kinesin Eg5: a transient and time-resolved fluorescence study.
2013,
Pubmed Muretta,
The structural kinetics of switch-1 and the neck linker explain the functions of kinesin-1 and Eg5.
2015,
Pubmed Myers,
Kinesin-5 regulates the growth of the axon by acting as a brake on its microtubule array.
2007,
Pubmed Nadar,
Kinesin-5 is essential for growth-cone turning.
2008,
Pubmed Roostalu,
Directional switching of the kinesin Cin8 through motor coupling.
2011,
Pubmed Rosenfeld,
Docking and rolling, a model of how the mitotic motor Eg5 works.
2005,
Pubmed Rosenfeld,
Stepping and stretching. How kinesin uses internal strain to walk processively.
2003,
Pubmed Ruhnow,
Tracking single particles and elongated filaments with nanometer precision.
2011,
Pubmed Schnitzer,
Statistical kinetics of processive enzymes.
1995,
Pubmed Scholey,
Structural basis for the assembly of the mitotic motor Kinesin-5 into bipolar tetramers.
2014,
Pubmed Shastry,
Interhead tension determines processivity across diverse N-terminal kinesins.
2011,
Pubmed Shastry,
Neck linker length determines the degree of processivity in kinesin-1 and kinesin-2 motors.
2010,
Pubmed Shimamoto,
Measuring Pushing and Braking Forces Generated by Ensembles of Kinesin-5 Crosslinking Two Microtubules.
2015,
Pubmed
,
Xenbase Sindelar,
An atomic-level mechanism for activation of the kinesin molecular motors.
2010,
Pubmed Skoufias,
S-trityl-L-cysteine is a reversible, tight binding inhibitor of the human kinesin Eg5 that specifically blocks mitotic progression.
2006,
Pubmed Svoboda,
Fluctuation analysis of motor protein movement and single enzyme kinetics.
1994,
Pubmed Talapatra,
The structure of the ternary Eg5-ADP-ispinesib complex.
2012,
Pubmed Valentine,
Force and premature binding of ADP can regulate the processivity of individual Eg5 dimers.
2009,
Pubmed Valentine,
Individual dimers of the mitotic kinesin motor Eg5 step processively and support substantial loads in vitro.
2006,
Pubmed Valentine,
To step or not to step? How biochemistry and mechanics influence processivity in Kinesin and Eg5.
2007,
Pubmed Varga,
Kinesin-8 motors act cooperatively to mediate length-dependent microtubule depolymerization.
2009,
Pubmed Verbrugge,
Novel ways to determine kinesin-1's run length and randomness using fluorescence microscopy.
2009,
Pubmed
