Spatiotemporal control of kinesin motor protein by photoswitches enabling selective single microtubule regulations

Kumar, K. R. Sunil; Amrutha, Ammathnadu S.; Tamaoki, Nobuyuki

doi:10.1039/c6lc01098a

Cited by 15 publications

(13 citation statements)

References 36 publications

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“…165 Similar systems with macroscopic components in the control loop have been repeatedly studied, but they clearly remove the autonomy of the molecular robot and turn the kinesin-microtubule system into a machine with remote control. 166–172 However, a gliding microtubule navigating through channels and around obstacles on a surface exhibits autonomous behavior similar to Walter’s tortoises. The gliding microtubule “chooses” a path which minimizes the bending of the microtubule.…”

Section: Engineering Autonomous Molecular Robotsmentioning

confidence: 99%

Non-equilibrium assembly of microtubules: from molecules to autonomous chemical robots

2017

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Biological systems have evolved to harness non-equilibrium processes from the molecular to the macro scale. It is currently a grand challenge of chemistry, materials science, and engineering to understand and mimic biological systems that have the ability to autonomously sense stimuli, process these inputs, and respond by performing mechanical work. New chemical systems are responding to the challenge and form the basis for future responsive, adaptive, and active materials. In this article, we describe a particular biochemical-biomechanical network based on the microtubule cytoskeletal filament – itself a non-equilibrium chemical system. We trace the non-equilibrium aspects of the system from molecules to networks and describe how the cell uses this system to perform active work in essential processes. Finally, we discuss how microtubule-based engineered systems can serve as testbeds for autonomous chemical robots composed of biological and synthetic components.

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Section: Engineering Autonomous Molecular Robotsmentioning

confidence: 99%

Non-equilibrium assembly of microtubules: from molecules to autonomous chemical robots

2017

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“…We used the same optical setup developed by our group earlier to translocate the microtubule and to manipulate its movement selectively. 29 Here we used only one wavelength of light, unlike our previous report where we used UV and visible wavelengths simultaneously. The schematic representation of the optical setup used is provided in the Supporting Information (Figure S13).…”

Section: Resultsmentioning

confidence: 99%

“…We recently reported the local regulation of kinesin motor activity through azobenzene-based photoswitches. 29 There, we selectively induced motility into a particular microtubule by irradiating the selected microtubule with one wavelength and the entire imaging area with the other wavelength. There, the irradiation of the entire imaging area with the second wavelength was essential to avoid the activation of motors present in the surrounding region by diffused active molecules.…”

Section: Discussionmentioning

confidence: 99%

“…27,28 With these two photoswitches, we achieved the local regulation of kinesin motor activity by simultaneous irradiation with UV and visible light using a simple, in-house built optical setup. 29 By reducing the light beam size, we were able to select a microtubule and manipulate its movement specifically. Though our method involved no surface patterning and prefunctionalization, optimization of the two light intensities to achieve an appropriate isomer ratio at the specific region of interest was challenging.…”

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confidence: 99%

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Targeted Activation of Molecular Transportation by Visible Light

et al. 2017

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Regulated transportation of nanoscale objects with a high degree of spatiotemporal precision is a prerequisite for the development of targeted molecular delivery. In vitro integration of the kinesin-microtubule motor system with synthetic molecules offers opportunities to develop controllable molecular shuttles for lab-on-a-chip applications. We attempted a combination of the kinesin-microtubule motor system with push-pull type azobenzene tethered inhibitory peptides (azo-peptides) through which reversible, spatiotemporal control over the kinesin motor activity was achieved locally by a single, visible wavelength. The fast thermal relaxation of the cis-isomers of azo-peptides offered us quick and complete resetting of the trans-state in the dark, circumventing the requirement of two distinct wavelengths for two-way switching of kinesin-driven microtubule motility. Herein, we report the manipulation of selected, single microtubule movement while keeping other microtubules at complete rest. The photoresponsive inhibitors discussed herein would help in realizing complex bionanodevices.

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“…One way to redirect microtubules locally is to change the local environment around microtubules. Tamaoki and coworkers (Amrutha, Kumar, Kikukawa, & Tamaoki, ; Kumar, Amrutha, & Tamaoki, ) designed light‐controlled microtubule transport mechanisms by introducing azobenzene‐derived nonnucleoside triphosphate as the substitute for ATP and azobenzene‐tethered peptide as kinesin inhibitor. Shining UV or visible light can switch photoisomerizable azobenzene between cis and trans forms to either trigger or block the microtubule gliding, respectively.…”

Section: Introductionmentioning

confidence: 99%

Local direction change of surface gliding microtubules

Pan

Choi

2019

Biotech & Bioengineering

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In vitro gliding assay, microtubule translocation by kinesin motor proteins on a surface, has been used as an engineering tool in analyte detection, molecular cargo transport, and other applications. Although controlling the moving direction is often necessary to realize these applications, current direction control methods focus largely on lithographic microfabrication of tracks or external fields on the microtubules. These methods are effective, but are relatively complicated. In addition, they cannot target particular microtubules without affecting others. In this study, we propose a facile approach that can make local direction changes for selected microtubules using a polystyrene particle as a circular motion center and a DNA double helix with streptavidin as a capture arm. The DNA arm captures a microtubule in the close proximity of the immobilized particle via biotin–streptavidin interaction and changes the moving direction ~10° on average. In contrast, no significant direction changes are observed other than random variations with streptavidin‐less DNA arms (normal distribution centered at 0°), similar to regular motility assay. The particle‐assisted local direction change scheme is compared with a flow field‐based ensemble method. The combination of flow and kinesin interactions with each microtubule exerts a force to change the direction, ultimately aligning it to the flow field, regardless of its initial direction. A simple model based on the force balance predicts the time needed for such an alignment. Overall, the particle‐based local scheme is distinct and different from ensemble methods such as crossflow that changes directions of all microtubules in the field, thus offering unique utility in engineering applications.

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Spatiotemporal control of kinesin motor protein by photoswitches enabling selective single microtubule regulations

Cited by 15 publications

References 36 publications

Non-equilibrium assembly of microtubules: from molecules to autonomous chemical robots

Non-equilibrium assembly of microtubules: from molecules to autonomous chemical robots

Targeted Activation of Molecular Transportation by Visible Light

Local direction change of surface gliding microtubules

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