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Visualization of Hydrophobic-, Hydrophilic-, and Charged Areas of Protein by Photochromic Cantilevers.

Research Project

Project/Area Number 10640384
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 物理学一般
Research InstitutionKanazawa University

Principal Investigator

ANDO Toshio  Kanazawa Univ., Department of Physics, Professor, 理学部, 教授 (50184320)

Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1998: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsProtein / Force Mapping / AFM / Photochromic dye / Cantilever / Hydrophobicity / Hydrophilicity / Electric Charge / AFM / 物性マッピング / 蛋白質表面物性 / 可視化
Research Abstract

Atomic force microscope (AFM) was first invented as an imaging tool for nanometer worlds. However, it can be used also as a high-sensitive force sensor, which allows us to obtain a 2D map of forces exerted between a cantilever probe and sample surface. Adhesive, repulsive, rupture, or non-contact forces between an AFM probe and sample can tell us physicochemical properties of the sample such as hydrophobicity, hydrophilicity and electric charges, depending on the surface property of the probe used. To specify the physicochemical property of sample clearly it is best to scan the sample with probes having different properties. Yet, it is quite difficult to scan the same area of sample after changing cantilevers. This has been a main reason why AFM has not been successfully used for mapping property of protein surface. To detour this problem we developed an alternative method. Instead of changing cantilevers, we change the surface property of a photochromic probe by irradiating near-UV li … More ght. We synthesized a photochromic dye, vinyl malachite green (VMG) whose structure can change from hydrophobic to charged form on irradiation. This dye was attached to the tip of cantilever probes covalently. We also invented a scanning method to map rupture forces within a time shorter than an ordinary force-curve method. Although it was shorter, it still took 30 min, resulting in suffering from disturbance by mechanical and electrical drifts of the AFM.After confirming the usefulness of the photochromic probe in identifying physicochemical property of sample, we invented an additional scanning method to obtain non-contact force map and topography simultaneously. This method shortened the scan time to 5 min (although it is not short enough yet). The positive charges on a basic protein, lysozyme, was successfully visualized by this method. Parallel to these studies, we have developed a high-speed AFM.It can ultimately solve the problem that the observation of force map requires much longer time than topography. The frame rate for topography observation reached 2.5 frames/sec, about 250-times faster than ordinary AFM apparatus. Although we have not tried to use this new AFM for force mapping, its speed promises a few seconds of force mapping, and therefore in the near future we may be able to obtain much better resolution of force map on protein surfaces. Less

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] I.Amitani, T.Sakamoto, T.Ando: "Link between the Enzymatic Kinetics and Mechanical Bahavior in an Actomyosin Motor."Biophysical Journal. 80. 379-397 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T.Sakamoto, I.Amitani, E.Yokota, T.Ando: "Direct Observation of Processive Movement by Individual Myosin V Molecules."Biochem.Biophys.Res.Commun.. 272. 586-590 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Adachi, K.Kinosita,Jr, T.Ando: "Single-fluorophore Imaging with an Unmodified Epifluorescence Microscope and Conventional Video Camera."Journal of Microscopy. 195. 125-132 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] I.Amitani, T.Sakamoto & T.Ando: "Link between the Enzymatic Kinetics and Mechanical Behavior in an Actomyosin Motor."Biophys.J. 80. 379-397 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T.Sakamoto, I.Amitani, E.Yokota & T.Ando: "Direct Observation of Processive Movement by Individual Myosin V Molecules."Biochem.Biophys.Res.Commun. 272. 586-590 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Adachi, K.Kinosita, Jr. & T.Ando: "Single-fluorophore Imaging with an Unmodified Epifluorescence Microscope and Conventional Video Camera."J.Microscopy. 195. 125-132 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Ando et al.: "Single-fluorophore imaging with an unmodified epi-fluorescence microscope and conventional video camera"Journal of Microscopy. 195. 125-132 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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