Kinesin Movement on Glutaraldehyde-Fixed Microtubules

Turner, David C.; Chang, Chun‐Yen; Fang, Kan; Cuomo, Peter; Murphy, Douglas B.

doi:10.1006/abio.1996.0422

Cited by 51 publications

(35 citation statements)

References 2 publications

(2 reference statements)

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“…This statistical analysis is important for applying motor proteins to quantitative and parallel cargo transport. However, the maximum velocity of 0.44 mm s À1 for kinesin-coated microspheres in our experiment is comparable to previous studies (Block et al, 1990;Svoboda et al, 1993), and is also consistent with velocities of cargo transported on glutaraldehyde-fixed microtubules (Doot et al, 2007;Turner et al, 1996). The maximum velocity of 0.80 mm s À1 for dynein-coated microspheres is also in agreement with results for single molecule assay (King and Schroer, 2000;Toba et al, 2006).…”

Section: Resultssupporting

confidence: 92%

“…Dynein-coated microspheres were transported on glutaraldehyde-fixed microtubules at maximum velocities similar to in vitro bead assay-based system. While it was reported that cross-linking of microtubules with glutaraldehyde does not affect kinesin motility (Doot et al, 2007;Turner et al, 1996), this is the first report for simultaneous kinesin and dynein motility on such ''cross-linked'' microtubules. Such results ensure that motility of two motor proteins on chemically immobilized microtubules can be utilized for bidirectional cargo nanotransport.…”

Section: Resultsmentioning

confidence: 65%

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Simultaneous and bidirectional transport of kinesin‐coated microspheres and dynein‐coated microspheres on polarity‐oriented microtubules

Yokokawa

Tarhan

Kon

et al. 2008

Biotech & Bioengineering

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Artificial nanotransport systems inspired by intracellular transport processes have been investigated for over a decade using the motor protein kinesin and microtubules. However, only unidirectional cargo transport has been achieved for the purpose of nanotransport in a microfluidic system. Here, we demonstrate bidirectional nanotransport by integrating kinesin and dynein motor proteins. Our molecular system allows microtubule orientation of either polarity in a microfluidic channel to construct a transport track. Each motor protein acts as a nanoactuators that transports microspheres in opposite directions determined by the polarity of the oriented microtubules: kinesin-coated microspheres move toward the plus end of microtubules, whereas dynein-coated microspheres move toward the minus end. We demonstrate both unidirectional and bidirectional transport using kinesin- and dynein-coated microspheres on microtubules oriented and glutaraldehyde-immobilized in a microfluidic channel. Tracking and statistical analysis of microsphere movement demonstrate that 87-98% of microspheres move in the designated direction at a mean velocity of 0.22-0.28 microm/s for kinesin-coated microspheres and 0.34-0.39 microm/s for dynein-coated microspheres. This bidirectional nanotransport goes beyond conventional unidirectional transport to achieve more complex artificial nanotransport in vitro.

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Section: Resultssupporting

confidence: 92%

Section: Resultsmentioning

confidence: 65%

Simultaneous and bidirectional transport of kinesin‐coated microspheres and dynein‐coated microspheres on polarity‐oriented microtubules

Yokokawa

Tarhan

Kon

et al. 2008

Biotech & Bioengineering

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“…This means that purified tubulin manipulation to produce microtubules is a relatively complex procedure. A treatment widely used to prevent microtubule disassembly by dilution and low temperature is a mild fixation with glutaraldehyde (45)(46)(47). 0.2% glutaraldehyde has been used to fix the cytoskeleton of Ptk2 cells, maintaining their ability to specifically bind Flutax-1 (24).…”

Section: Stabilized Taxoid Binding Sites Provided By Cross-linkedmentioning

confidence: 99%

Molecular Recognition of Taxol by Microtubules

Díaz¹,

Strobe²,

Engelborghs³

et al. 2000

Journal of Biological Chemistry

117

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“…The critical issue of microtubule deactivation has also been addressed through stabilization with taxol [96] or glutaraldehyde. [97] …”

Section: Surface Effectsmentioning

confidence: 99%

Protein Linear Molecular Motor-Powered Nanodevices

Bakewell

Nicolau

2007

Aust. J. Chem.

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Myosin-actin and kinesin-microtubule linear protein motor systems and their application in hybrid nanodevices are reviewed. Research during the past several decades has provided a wealth of understanding about the fundamentals of protein motors that continues to be pursued. It has also laid the foundations for a new branch of investigation that considers the application of these motors as key functional elements in laboratory-on-a-chip and other micro/nanodevices. Current models of myosin and kinesin motors are introduced and the effects of motility assay parameters, including temperature, toxicity, and in particular, surface effects on motor protein operation, are discussed. These parameters set the boundaries for gliding and bead motility assays. The review describes recent developments in assay motility confinement and unidirectional control, using micro-and nano-fabricated structures, surface patterning, microfluidic flow, electromagnetic fields, and self-assembled actin filament/microtubule tracks. Current protein motor assays are primitive devices, and the developments in governing control can lead to promising applications such as sensing, nano-mechanical drivers, and biocomputation.

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Kinesin Movement on Glutaraldehyde-Fixed Microtubules

Cited by 51 publications

References 2 publications

Simultaneous and bidirectional transport of kinesin‐coated microspheres and dynein‐coated microspheres on polarity‐oriented microtubules

Simultaneous and bidirectional transport of kinesin‐coated microspheres and dynein‐coated microspheres on polarity‐oriented microtubules

Molecular Recognition of Taxol by Microtubules

Protein Linear Molecular Motor-Powered Nanodevices

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