Biomolecular linear motors confined to move upon micro-patterns on glass

Yoshida, Yumi; Yokokawa, Ryuji; Suzuki, Hiroaki; Atsuta, Kyoko; Fujita, Hiroyuki; Takeuchi, Shoji

doi:10.1088/0960-1317/16/8/015

Cited by 24 publications

(17 citation statements)

References 15 publications

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“…MTs moved between these states in a dynamic manner, as indicated by the arrows, and eventually reached the final three conditions: detaching from the surface, returning to the original region, and passing the boundary to the other region. The third classification has not been seen in previous studies in which only kinesin was involved2728. Whether MTs approached the boundary from dynein- or kinesin-coated regions, the majority (64.3%, Fig.…”

Section: Resultsmentioning

confidence: 74%

Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface

Ikuta

Kamisetty

Shintaku

et al. 2014

Sci Rep

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Intracellular cargo is transported by multiple motor proteins. Because of the force balance of motors with mixed polarities, cargo moves bidirectionally to achieve biological functions. Here, we propose a microtubule gliding assay for a tug-of-war study of kinesin and dynein. A boundary of the two motor groups is created by photolithographically patterning gold to selectively attach kinesin to the glass and dynein to the gold surface using a self-assembled monolayer. The relationship between the ratio of two antagonistic motor numbers and the velocity is derived from a force-velocity relationship for each motor to calculate the detachment force and motor backward velocity. Although the tug-of-war involves >100 motors, values are calculated for a single molecule and reflect the collective dynein and non-collective kinesin functions when they work as a team. This assay would be useful for detailed in vitro analysis of intracellular motility, e.g., mitosis, where a large number of motors with mixed polarities are involved.

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Section: Resultsmentioning

confidence: 74%

Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface

Ikuta

Kamisetty

Shintaku

et al. 2014

Sci Rep

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“…The direction of microtubule movement has been controlled by passive methods such as chemically modified or patterned surfaces (Cheng et al, 2005;Clemmens et al, 2003b;Dennis et al, 1999;Hess et al, 2002;Yoshida et al, 2006), and microfabricated substrates (Cheng et al, 2005;Clemmens et al, 2003aClemmens et al, ,b, 2004Hess et al, , 2002Hess et al, , 2003Hiratsuka et al, 2001;Jia et al, 2004;Moorjani et al, 2003;van den Heuvel et al, 2005). Kinesin molecules are selectively absorbed on a substrate to confine microtubule motility in a certain area, or microtubules are guided by microfabricated tracks.…”

Section: Introductionmentioning

confidence: 99%

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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“…A flow cell with micro-patterned ssDNA spots designed to be unloading sites was prepared applying the poly-para-xylylene (parylene) peel-off process [11]. Parylene was coated on a glass substrate to form a thin film by vapor deposition (Figure 2 (1)), and then a patterned mask (arrayed holes with 20 µm-diameter) was coated on the parylene film (Figure 2 (2)).…”

Section: Fabrication Of An Ssdna Arraymentioning

confidence: 99%

Biomolecular motor-based cargo transporters with loading/unloading mechanisms on a micro-patterned DNA array

Hiyama

Takeuchi

Gojo

et al. 2008

2008 IEEE 21st International Conference on Micro Electro Mechanical Systems

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This paper describes research in creating autonomous cargo transporters that load, transport, and unload cargo molecules using the machinery in living cells. By exploiting biomolecular motor-based motility and DNA hybridization, we constructed autonomous cargo transporters and demonstrated that kinesin-driven microtubules can selectively load and transport cargoes toward micropatterned DNA spots designed to be unloading sites and selectively unload the transported cargoes at the DNA spots specified by particular DNA base sequences. These autonomous operations may help create highly miniaturized on-chip-systems such as molecular sorters and sensors, and may also provide an alternative technology to pressuredriven or electrokinetic flow-based microfluidic devices.

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Biomolecular linear motors confined to move upon micro-patterns on glass

Cited by 24 publications

References 15 publications

Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface

Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface

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

Biomolecular motor-based cargo transporters with loading/unloading mechanisms on a micro-patterned DNA array

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