Hybrid nanotransport system by biomolecular linear motors

We have demonstrated a novel micro/nanotransport system using biomolecular motors driven by adenosine triphosphate (ATP). For the driving mechanism, microtubule-kinesin system, which is one of the linear biomolecular motor systems was investigated. ATP dissolved in an aqueous condition is hydrolyzed to adenosine diphosphate (ADP) to energize the bionanoactuators in this mechanism. This means the system does not require an external electrical or mechanical energy source. Therefore, a purely chemical system which is similar to the in vivo transport will be realized. This paper reports some fundamental studies to integrate biomaterials and MEMS. The microtubules, or rail molecules, were patterned on a glass substrate with poly(dimethyl siloxane) (PDMS) using a regular soft lithography technique. Microbeads (320 nm in diameter) and a micromachined structure (2/spl times/3 /spl mu/m, 2 /spl mu/m in thickness) coated with kinesin molecules were transported along the microtubules at an average speed of 476/spl plusmn/56 and 308 nm/s, respectively. While ATP injection activated the transport system we have also managed to provide repetitive on/off control using hexokinase as an inhibitor. For the minimum response time in the repetitive control, the optimized concentration for ATP was 10/sup 2/ /spl mu/M and 10/sup 3/ U/L for hexokinase.

[1]  S. Ishiwata,et al.  Temperature dependence of force, velocity, and processivity of single kinesin molecules. , 2000, Biochemical and biophysical research communications.

[2]  T Kanayama,et al.  Controlling the direction of kinesin-driven microtubule movements along microlithographic tracks. , 2001, Biophysical journal.

[3]  Hiroyasu Itoh,et al.  Resolution of distinct rotational substeps by submillisecond kinetic analysis of F1-ATPase , 2001, Nature.

[4]  Ronald D. Vale,et al.  Single-molecule analysis of kinesin motility reveals regulation by the cargo-binding tail domain , 1999, Nature Cell Biology.

[5]  Toshio Yanagida,et al.  Direct observation of single kinesin molecules moving along microtubules , 1996, Nature.

[6]  H. Neves,et al.  The art and science of engineering hybrid living/non-living mechanical devices , 2002, Technical Digest. MEMS 2002 IEEE International Conference. Fifteenth IEEE International Conference on Micro Electro Mechanical Systems (Cat. No.02CH37266).

[7]  H. Fujita,et al.  On/off control of biomolecular motors in a microfluidic device , 2003, TRANSDUCERS '03. 12th International Conference on Solid-State Sensors, Actuators and Microsystems. Digest of Technical Papers (Cat. No.03TH8664).

[8]  F. Arai,et al.  Pinpoint injection of micro tools using dielectrophoresis and hydrophobic surface for minimally invasive separation of microbe , 2002, Technical Digest. MEMS 2002 IEEE International Conference. Fifteenth IEEE International Conference on Micro Electro Mechanical Systems (Cat. No.02CH37266).

[9]  Beads in Biochemical Microfluidics , 2002 .

[10]  Toshio Yanagida,et al.  A single myosin head moves along an actin filament with regular steps of 5.3 nanometres , 1999, Nature.

[11]  Jin-Woo Choi,et al.  A new magnetic bead-based, filterless bio-separator with planar electromagnet surfaces for integrated bio-detection systems , 2000 .

[12]  C. Ahn,et al.  An on-chip magnetic bead separator using spiral electromagnets with semi-encapsulated permalloy. , 2001, Biosensors & bioelectronics.

[13]  M. Baum,et al.  Effect of temperature on kinesin‐driven microtubule gliding and kinesin ATPase activity , 2000, FEBS letters.

[14]  K. Greulich,et al.  Laser manipulation and UV induced single molecule reactions of individual DNA molecules , 1996 .

[15]  Loren L Looger,et al.  Control of a biomolecular motor-powered nanodevice with an engineered chemical switch , 2002, Nature materials.