Harnessing the motion of motor proteins for transporting biomolecules requires directional control. The chemo-mechanical energy conversion of motor proteins is highly efficient and once directional control is achieved, a broad range of applications from delivery, assembly, sorting and detection to micro/nano-engines and energy transduction can be realized for a whole new generation of hybrid bio-devices. The effort is to develop a methodology that will have applicability on the transport of specific proteins from one location to a targeted location for further manipulation for security, health or environmental applications. The general approach to be developed is based on functionalized micro/nano-sized patterning pathways on an inorganic substrate. Micro-patterning will be developed along with localized flow fields to arrange and align F-actin on selected surfaces with one structural polarity attached to the substrate. This approach will give unidirectional movement of myosin coated shuttle particles such as beads, nanowires and nanotubes, where specific cargo can be bound and transported.
Intellectual merit: The understanding and ability to develop transportation systems at nano-scale is of paramount importance and will enable the capability to explore and engineer this new frontier. The proposed research aims to develop transport systems suitable for cargo delivery which are propelled by motor proteins. Engineered pathways will be accomplished by means of orientated assembly of actin protein, selective molecular micropatterning and photoreactive polymer synthesis. The knowledge to be gained through this research includes generation of specific flow fields and hybrid device integration. The broader understanding gained through this work will lay foundation for the future applications of autonomous transport and actuation systems, whether biological or synthetic in nature at nano-scale. Broader impact: The proposed research effort seeks to establish the underlying framework for cargo transport powered by nano-scale biomolecular motors from within a microelectronic/chip environment. The proposed work will examine fundamental properties of nano pathways and tracks for transport purposes. Given the inevitable integration of electrical and biological components in hybrid nano-systems, the knowledge gained about the relationship between the protein-bead-cargo-myosin shuttle system in micro/nano-tracks motility assays and their viability, is valuable to a wide range of research fields beyond this fundamental study. This work is anticipated to have an impact on engineering, physiological and biological research, as a working model of physiological molecular transport.
A challenge associated with the promise of nanotechnology is training the workforce needed to support its advancement and implementation. The inherent interdisciplinary nature of this training is not well addressed by traditional academic, discipline-based programs. The activities of this proposal offer opportunities for integrating students' educational experience across diverse areas including microfabrication, biochemistry, nanotechnology, microfluidics, physics, and chemistry. The educational outreach program of this project will serve to motivate the next generation of cross-disciplinary scientists by introducing high school students and freshmen to cutting edge research in laboratories, and by providing undergraduate researchers, especially underrepresented groups including women and minorities, the opportunity to conduct cutting edge, genuine research in a state-of-the-art setting.
The understanding and ability to develop transportation systems at nano-scale is of paramount importance and will enable the capability to explore and engineer this new frontier. Harnessing the motion of motor proteins for transporting biomolecules requires directional control. The chemo-mechanical energy conversion of motor proteins is highly efficient and once directional control is achieved, a broad range of applications from delivery, assembly, sorting and detection to micro/nano-engines and energy transduction can be realized for a whole new generation of hybrid bio-devices (Figure 1). This research aimed to develop transport systems suitable for cargo delivery which are propelled by motor proteins. Engineered pathways were accomplished by means of orientated assembly of actin protein, selective molecular micropatterning and photoreactive polymer synthesis. Here we have developed a methodology that will have applicability on the transport of specific particles and/or proteins from one location to a targeted location for further manipulation for security, health or environmental applications. The general approach that was developed is based on functionalized micro/nano-sized patterning pathways on an inorganic substrate. Micro-patterning has been developed along with localized flow fields to arrange and align F-actin on selected surfaces with one structural polarity attached to the substrate. This approach has given unidirectional movement of myosin coated shuttle particles such as beads (Figure 2), nanowires and nanotubes, where specific cargo can be bound and transported (Figure 3). The successful implementation of oriented surface patterns the actin tracks were integrated into microscale channels and further cross-linked with fascin to sustain the streptavidin layer required for patterning. The transport of HMM-coated particles on these fascin-modified arrays was identical to the data obtained with the original arrays devoid of fascin. In addition, no particle movement was observed in areas that were not patterned. As shown in Figure 4, this approach yielded 100 μm wide patterned arrays. These well-oriented F-actin arrays provided unidirectional transport of heavy meromyosin (HMM) -coated particles over distances of several hundred micrometers as shown Figure 5. The activity of HMM is greatly influenced by adsorption behavior on the surface. Particles were coated with PLL for bead assays. HMM-coated particles in contact with the arrays moved along the arrays. All particles moving on the arrays showed unidirectional movement toward the end of the flow cell where the flow solution had been introduced to lay down F-actin. The particle size limited the number of HMM heads on the area in contact with F-actin arrays. This caused the particles to travel at different velocities that varied according to the particle size. Detailed analysis of the HMM-coated particles on fascin-crosslinked actin arrays revealed that the actin filament was modified by the myosin motors so that further transport was hindered. After the HMM-coated particles passed over a track, it appeared to disconnect from the surface and was released in the medium as shown Figure 6. These preliminary results provide direct evidence to support this hypothesis as well as insight into the mechanism of myosin II induced depolymerization of actin. In summary this research work examined fundamental properties of nano pathways and tracks for transport purposes. Given the inevitable integration of electrical and biological components in hybrid nano-systems, the knowledge gained about the relationship between the protein-bead-cargo-myosin shuttle system in micro/nano-tracks motility assays and their viability, is valuable to a wide range of research fields beyond this fundamental study. This work is anticipated to have an impact on engineering, physiological and biological research, as a working model of physiological molecular transport. A challenge associated with the promise of nanotechnology is training the workforce needed to support its advancement and implementation. The inherent interdisciplinary nature of this training is not well addressed by traditional academic, discipline-based programs. The activities of this research have offered opportunities for integrating students’ educational experience across diverse areas including microfabrication, biochemistry, nanotechnology, microfluidics, physics, and chemistry. The educational outreach program of this research has served and motivated the next generation of cross-disciplinary scientists by introducing high school students and freshmen to cutting edge research in our laboratories, and we provided undergraduate researchers, especially underrepresented groups including women and minorities, the opportunity to conduct cutting edge, genuine research in a state-of-the-art setting. The knowledge to be gained through this research includes generation of specific flow fields and hybrid device integration. The broader understanding gained through this work will lay foundation for the future applications of autonomous transport and actuation systems, whether biological or synthetic in nature at nano-scale.
