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A kinesin motor in a force-producing conformation.
Abstract
BACKGROUND: Kinesin motors hydrolyze ATP to produce force and move along microtubules,
converting chemical energy into work by a mechanism that is only poorly understood.
Key transitions and intermediate states in the process are still structurally uncharacterized,
and remain outstanding questions in the field. Perturbing the motor by introducing
point mutations could stabilize transitional or unstable states, providing critical
information about these rarer states. RESULTS: Here we show that mutation of a single
residue in the kinesin-14 Ncd causes the motor to release ADP and hydrolyze ATP faster
than wild type, but move more slowly along microtubules in gliding assays, uncoupling
nucleotide hydrolysis from force generation. A crystal structure of the motor shows
a large rotation of the stalk, a conformation representing a force-producing stroke
of Ncd. Three C-terminal residues of Ncd, visible for the first time, interact with
the central beta-sheet and dock onto the motor core, forming a structure resembling
the kinesin-1 neck linker, which has been proposed to be the primary force-generating
mechanical element of kinesin-1. CONCLUSIONS: Force generation by minus-end Ncd involves
docking of the C-terminus, which forms a structure resembling the kinesin-1 neck linker.
The mechanism by which the plus- and minus-end motors produce force to move to opposite
ends of the microtubule appears to involve the same conformational changes, but distinct
structural linkers. Unstable ADP binding may destabilize the motor-ADP state, triggering
Ncd stalk rotation and C-terminus docking, producing a working stroke of the motor.
Type
Journal articleSubject
Adenosine DiphosphateAdenosine Triphosphate
Amino Acid Sequence
Amino Acid Substitution
Animals
Binding Sites
Crystallography, X-Ray
Drosophila Proteins
Drosophila melanogaster
Hydrolysis
Kinesin
Kinetics
Microtubules
Models, Molecular
Mutation
Protein Structure, Secondary
Rotation
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https://hdl.handle.net/10161/4362Published Version (Please cite this version)
10.1186/1472-6807-10-19Publication Info
Heuston, Elisabeth; Bronner, C Eric; Kull, F Jon; & Endow, Sharyn A (2010). A kinesin motor in a force-producing conformation. BMC Struct Biol, 10. pp. 19. 10.1186/1472-6807-10-19. Retrieved from https://hdl.handle.net/10161/4362.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Sharyn Anne Endow
Professor of Cell Biology
Research in my laboratory focuses on spindle and chromosome dynamics and the mechanisms
that ensure proper chromosome transmission and inheritance in dividing cells. Work
in my laboratory and others over the past 5-10 years has identified molecular motor
proteins as the force-generating proteins underlying movements of the spindle and
chromosomes during cell division. Much of our current effort is directed towards understanding
the mechanism of motor function, including the molecular basis of mo

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