| Project/Area Number |
06558099
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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 | 試験 |
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| Research Field |
Biophysics
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| Research Institution | Kanazawa University |
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Principal Investigator |
ANDO Toshio Kanazawa University, Science, Professor, 理学部, 教授 (50184320)
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| Co-Investigator(Kenkyū-buntansha) |
TAKENOBU Takayoshi Olympus, 2nd R/D,Researcher, 第2開発部, 研究員
HAYASHI Yoshiaki Olympus, 2nd R/D,Chief, 第2開発部, 係長
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1994: ¥8,700,000 (Direct Cost: ¥8,700,000)
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| Keywords | Atomic Force Microscope / AFM / High-speed Scanner / High-speed Scanning / Protein / Bi-molecular Interaction / Myosin Heads / 高分解能像 / 分子間力 / カンチレバ- / 走査型プローブ顕微鏡 / 1分子力学計測 / 分子間力顕微鏡 / アクトミオシンモーター / ビデオレートAFM / 生物研究用AFM |
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| Research Abstract |
(a) We manufactured an atomic force microscope that was able to be scanned faster and less susceptible to drift than ordinary AFM apparatuses. This AFM was attached to an epiluminescence fluorescence microscope. To show the high performance of our AFM apparatus we tried to obtain high resolution images of myosin heads (subfragment-1) in aqueous solution. The images revealed fine structures that was comparable to those in an atomic model of S1 derived from x-ray crystallography of S1. In the AFM images we were able to see the ATP binding site as well as the large cleft at the tip of S1. These results proved that our AFM apparatus was on the level of the greatest in the world.
(b) We performed a study preparatory to the development of a high-speed AFM in which one image would be acquired within 30msec. We designed and manufactured a high-speed scanner and attained high-speed imaging where one image was obtained within 150msec. By way of experiment we produced an AFM that employed a displacemant detection system suitable for high-speed imaging. We also examinds methods for preparing cantilevers that were suitable for a high-speed AFM.From these preparative studies we were able to secure a scaffolding for the development of the video-rate AFM.
(c) To measure bimolecular interaction force at a truly single molecular level we developed a method to capture a single molecule of protein at the apex of a cantilever tip. Using this method we succeeded in quantifying the force field exerted between a single molecule of heavy meromyosin (HMM) and actin. Using this new technology we also discovered that ATP binding to HMM heads induced vibratile structural changes in HMM heads.
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