2005 Fiscal Year Final Research Report Summary
Novel Single Bio-Molecule Measurement using Micro/Nano Machining Technology
| Project/Area Number |
14205035
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Intelligent mechanics/Mechanical systems
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| Research Institution | The University of Tokyo |
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Principal Investigator |
KIM Beomjoon The University of Tokyo, Institute of Industrial Science, Associate Professor, 生産技術研究所, 助教授 (60334356)
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| Co-Investigator(Kenkyū-buntansha) |
NOJI Hiroyuki Osaka University, The Institute of Scientific and Industrial Research, Professor, 産業科学研究所, 教授 (00343111)
TAKEUCHI Shoji The University of Tokyo, Institute of Industrial Science, Associate Professor, 生産技術研究所, 助教授 (90343110)
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| Project Period (FY) |
2002 – 2005
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| Keywords | Bio-molecular motor / Micro / Nano machine / Nano machining technology / MEMS / Biophysics / Single molecule measurement / Nano patterning |
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| Research Abstract |
F1-ATPase is the smallest known rotary motor, and it rotates in an anticlockwise direction as it hydrolyses ATP. Single-molecule experiments point towards three catalytic events per turn, in agreement with the molecular structure of the complex. The physiological function of F1 is ATP synthesis. In the ubiquitous F0F1 complex, this energetically uphill reaction is driven by F0, the partner motor of F1, which forces the backward (clockwise) rotation of F1, leading to ATP synthesis. Here, we have devised an experiment combining single-molecule manipulation and microfabrication techniques to measure the yield of this mechanochemical transformation. Single F1 molecules were enclosed in femtolitre-sized hermetic chambers and rotated in a clockwise direction using magnetic tweezers. When the magnetic field was switched off, the F1 molecule underwent anticlockwise rotation at a speed proportional to the amount of synthesized ATP. We provide here direct evidence that F1 is designed to tightly
… More
couple its catalytic reactions with the mechanical rotation. Our results suggest that the 1-subunit has an essential function during ATP synthesis. Moreover, we propose a novel technique for patterning active proteins on a glass substrate using a perforated polydimethylsiloxane (PDMS) sheet-sieve. The sieve, which has tapering holes, is fabricated by spin-coating PDMS on a pyramidal-shaped mold. By means of this sieve, FITC (fluorescent isothiocyanate, bovine)-albumin was successfully spotted in an array. The patterned spots were perfectly isolated, which eliminates the problem of non-specific binding of proteins to undesired areas. To show that proteins maintained their activity after the patterning, we used F1-ATPase biomolecular motors ; their activity can easily be verified by observing their rotary motion after patterning. Selective patterning with three kinds of fluorescent micro beads indicated the possibility of patterning of different proteins on the same substrate by using the shadow mask. Less
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Research Products
(14 results)
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[Journal Article] Temperature distrivution measurement on microfabricated thermodevice for single biomolecular observation using fluorescent dye2006
Author(s)
Arata, F.H., Low, P., Ishizuka, K., Bergaud, C., Kim, B.J., Noji, H., Fujita, H.
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Journal Title
Sensors and Actuators B : Chemical (In press)
Description
「研究成果報告書概要(和文)」より
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[Journal Article] Activation of pausing F1 motor by external force.2005
Author(s)
Hirono-Hara, Y., Ishizuka, K., Kinosita, K, Jr., Yoshida, M., Noji, H.
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Journal Title
Proc. Natl. Acad. Scie. U.S.A. 102
Pages: 4288-93
Description
「研究成果報告書概要(和文)」より
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[Journal Article] Highly coupled ATP synthesis by F1-ATPase single molecules2005
Author(s)
Rondelez, Y., Tresset, G., Nakashima, T., Kato-Yamada, Y., Fujita, H., Takeuchi, S., Noji, H.
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Journal Title
Description
「研究成果報告書概要(和文)」より
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