Kinesin processivity is gated by phosphate release

Kinesin-1 is a dimeric motor protein, central to intracellular transport, that steps hand-over-hand toward the microtubule (MT) plus-end, hydrolyzing one ATP molecule per step. Its remarkable processivity is critical for ferrying cargo within the cell: over 100 successive steps are taken, on average...

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Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS 2014-09, Vol.111 (39), p.14136-14140
Hauptverfasser: Milic, Bojan, Andreasson, Johan O. L., Hancock, William O., Block, Steven M.
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container_issue 39
container_start_page 14136
container_title Proceedings of the National Academy of Sciences - PNAS
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creator Milic, Bojan
Andreasson, Johan O. L.
Hancock, William O.
Block, Steven M.
description Kinesin-1 is a dimeric motor protein, central to intracellular transport, that steps hand-over-hand toward the microtubule (MT) plus-end, hydrolyzing one ATP molecule per step. Its remarkable processivity is critical for ferrying cargo within the cell: over 100 successive steps are taken, on average, before dissociation from the MT. Despite considerable work, it is not understood which features coordinate, or “gate,” the mechanochemical cycles of the two motor heads. Here, we show that kinesin dissociation occurs subsequent to, or concomitant with, phosphate (P ᵢ) release following ATP hydrolysis. In optical trapping experiments, we found that increasing the steady-state population of the posthydrolysis ADP·P ᵢ state (by adding free P ᵢ) nearly doubled the kinesin run length, whereas reducing either the ATP binding rate or hydrolysis rate had no effect. The data suggest that, during processive movement, tethered-head binding occurs subsequent to hydrolysis, rather than immediately after ATP binding, as commonly suggested. The structural change driving motility, thought to be neck linker docking, is therefore completed only upon hydrolysis, and not ATP binding. Our results offer additional insights into gating mechanisms and suggest revisions to prevailing models of the kinesin reaction cycle. Significance Kinesin-1 is a motor protein central to intracellular transport. Prevailing models of the kinesin mechanochemical cycle—which invoke docking of the neck linker domain upon ATP binding—fail to explain the remarkable processivity of kinesin, which represents a competition between dissociation from the microtubule and continuation of the stepping cycle. We show that kinesin dissociation, which characterizes the end of a processive run, is gated by phosphate release following ATP hydrolysis. The structural change driving kinesin motility, likely neck linker docking, is therefore completed only upon hydrolysis. Our results offer insights into gating mechanisms and necessitate revisions to existing models of the kinesin cycle.
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Prevailing models of the kinesin mechanochemical cycle—which invoke docking of the neck linker domain upon ATP binding—fail to explain the remarkable processivity of kinesin, which represents a competition between dissociation from the microtubule and continuation of the stepping cycle. We show that kinesin dissociation, which characterizes the end of a processive run, is gated by phosphate release following ATP hydrolysis. The structural change driving kinesin motility, likely neck linker docking, is therefore completed only upon hydrolysis. 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subjects Adenosine triphosphatase
adenosine triphosphate
Adenosine Triphosphate - metabolism
Animals
Binding sites
Biochemistry
Biological Sciences
Biophysical Phenomena
Chemical reactions
Dimers
dissociation
Drosophila Proteins - chemistry
Drosophila Proteins - genetics
Drosophila Proteins - metabolism
Freight
Hydrolysis
kinesin
Kinesin - chemistry
Kinesin - genetics
Kinesin - metabolism
Kinetics
Microtubules
Models, Biological
molecular motor proteins
Molecular Motor Proteins - chemistry
Molecular Motor Proteins - genetics
Molecular Motor Proteins - metabolism
Molecules
Motors
Nucleotides
Optical Tweezers
Phosphates
Phosphates - metabolism
Protein Structure, Quaternary
Proteins
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
title Kinesin processivity is gated by phosphate release
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