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Structural analysis of the motor protein prestin

Research Project

Project/Area Number 15086202
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Science and Engineering
Research InstitutionTohoku University

Principal Investigator

WADA Hiroshi  Tohoku University, Department of Bioengineering and Robotics, Professor (30111264)

Co-Investigator(Kenkyū-buntansha) KUMAGAI Izumi  Tohoku University, Department of Bimolecular Engineering, Professor (10161689)
IKEDA Katsuhisa  Juntendo University School of Medicen, Department of Otorhinolaryngology, Professor (70159614)
Project Period (FY) 2003 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥41,000,000 (Direct Cost: ¥41,000,000)
Fiscal Year 2006: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2005: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2004: ¥22,100,000 (Direct Cost: ¥22,100,000)
Fiscal Year 2003: ¥11,100,000 (Direct Cost: ¥11,100,000)
KeywordsOuter hari cell / Protein motor / Prestin / Atomic force microscopy / Imaging
Research Abstract

The aim of this project was to clarify the motor protein prestin, whose conformational change has been indicated to induce an elongation and contraction of outer hair cells in the inner ear. To this end, first, an attempt was made to observe the plasma membrane of prestin-transfected Chinese hamster ovary (CHO) cells and that of untransfected CHO cells by atomic force microscopy (AFM). As a result, particle-like structures probably indicating membrane proteins were recognized in the plasma membranes of both cells, although there were no distinctive differences between the two. Analysis of the shape and size of the observed structures showed that there were more particle-like structures with a diameter of 8-12 nm in the plasma membrane of the prestin-transfected CHO cells than in that of the untransfected CHO cells, suggesting that these particle-like structures are possibly prestin molecules.
The second endeavor was to more specifically detect prestin molecules in the plasma membrane of the prestin-transfected CHO cells to clarify the structure of prestin in more detail. For this purpose, an experimental approach combining AFM with anti-prestin antibody conjugated with quantum dots (Qdots), used as topographic surface markers, was carried out to detect individual prestin molecules. As Qdots are hard material with a diameter of 8 nm, they can be easily seen by AFM. In addition, as prestin may exist near Qdots, detection and observation of prestin molecules can be performed. As a result of imaging by AFM of the prestin-transfected CHO cells labeled with Qdots, squarish ring-like structures, each with four peaks and one valley at its center, were observed in the vicinity of the Qdots, indicating that prestin forms a tetramer in the plasma membranes of the prestin-transfected CHO cells.

Report

(5 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (54 results)

All 2007 2006 2005 2004 2003 Other

All Journal Article (28 results) (of which Peer Reviewed: 10 results) Presentation (16 results) Book (7 results) Remarks (1 results) Publications (2 results)

  • [Journal Article] Effects of heat stress on filamentous actin and prestin of outer hair cells in mice2007

    • Author(s)
      Yoko Kitsunai
    • Journal Title

      Brain Research 1177

      Pages: 47-58

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Recent findings on our auditory system : it is highly sensitive owing to the motility of sensory cells2007

    • Author(s)
      Hiroshi Wada
    • Journal Title

      International Journal of Acoustics and Vibration 12

      Pages: 75-82

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Biomechanics at Micro- and Nanoscale Levels2007

    • Author(s)
      Hiroshi Wada
    • Journal Title

      World Scientific, Singapore 4

      Pages: 3-13

    • NAID

      110006649726

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Medical Bio, Biomechanics at Micro- and Nanoscale and its application for medicine2007

    • Author(s)
      T. Kouno (ed. )
    • Journal Title

      Ohmsha, Japan

      Pages: 24-31

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Bio Medical Engineering, Biomechanics at Micor- and Nanoscale2007

    • Author(s)
      J. Ando (ed. )
    • Journal Title

      Japan Society of Medical and Biological Engineering 46(1)

      Pages: 1-4

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Effects of heat stress on filamentous actin and prestin of outer hair cells in mice2007

    • Author(s)
      Yoko Kitsunai, et al.
    • Journal Title

      Brain Research 1177

      Pages: 47-58

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Imaging by atomic force microscopy of the plasma membrane of prestin-transfected Chinese hamster ovary cells2006

    • Author(s)
      Michio Murakoshi
    • Journal Title

      Journal of the Association for Research in Otolaryngology 7

      Pages: 267-278

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Effects of heat stress on Young's modulus of outer hair cells in mice2006

    • Author(s)
      Michio Murakoshi
    • Journal Title

      Brain Research 1107

      Pages: 121-130

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Local mechanical properties of mouse outer hair cells : Atomic force microscopic study2006

    • Author(s)
      Michio Murakoshi
    • Journal Title

      Auris Nasus Larynx 33

      Pages: 149-157

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Imaging by atomic force microscopy of the plasma membrane of prestin-transfected Chinese hamster ovary cells2006

    • Author(s)
      Michio Murakoshi, et al.
    • Journal Title

      Journal of the Association for Research in Otolaryngology 7

      Pages: 267-278

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Effects of heat stress on Young's modulus of outer hair cells in mice2006

    • Author(s)
      Michio Murakoshi, et al.
    • Journal Title

      Brain Research 1107

      Pages: 121-130

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Local mechanical properties of mouse outer hair cells : atomic force microscopic study2006

    • Author(s)
      Michio Murakoshi, et al.
    • Journal Title

      Auris Nasus Larynx 33

      Pages: 149-157

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Biomechanics at Micro- and Nanoscale Levels2006

    • Author(s)
      Hiroshi Wada
    • Journal Title

      World Scientific, Singapore 2

      Pages: 3-15

    • NAID

      110006649726

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Analysis of the dynamic behavior of the inner hair cell stereocilia by the finite element method2006

    • Author(s)
      Toshihiro Matsui
    • Journal Title

      JSME International Journal 49C

      Pages: 828-836

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Local Mechanical Properties of Mouse Outer Hair Cells : Atomic Force Microscopic Study2006

    • Author(s)
      Michio Murakoshi
    • Journal Title

      Auris Nasus Larynx (in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Effects of mutation in the conserved GTSRH sequence of the motor protein prestin on its characteristics2005

    • Author(s)
      Shun Kumano
    • Journal Title

      JSME International Journal 48C

      Pages: 403-410

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Construction of an expression system for the motor protein prestin in Chinese hamster ovary cells2005

    • Author(s)
      Koji Iida
    • Journal Title

      Hearing Research 205

      Pages: 262-270

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2005 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Effects of mutation in the conserved GTSRH sequence of the motor protein prestin on its characteristics2005

    • Author(s)
      Shun Kumano, et al.
    • Journal Title

      JSME International Journal 48C

      Pages: 403-410

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Construction of an expression system for the motor protein prestin in Chinese hamster ovary cells2005

    • Author(s)
      Koji Iida, et al.
    • Journal Title

      Hearing Research 205

      Pages: 262-270

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Biomechanics at Micro- and Nanoscale Levels2005

    • Author(s)
      Hiroshi Wada
    • Journal Title

      World Scientific, Singapore 1

      Pages: 3-15

    • NAID

      110006649726

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Effects of Mutation in the Conserved GTSRH Sequence of the Motor Protein Prestin on Its Characteristics2005

    • Author(s)
      Shun Kumano
    • Journal Title

      JSME International Journal Series C 48

      Pages: 403-410

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Relationship between fluorescence intensity of GFP and the expression level of prestin in a prestin-expressing Chinese hamster ovary cell line2004

    • Author(s)
      Koji Iida
    • Journal Title

      JSME International Journal 47C

      Pages: 970-976

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2004 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Imaging of the cortical cytoskeleton of guinea pig outer hair cells using atomic force microscopy2004

    • Author(s)
      Hiroshi Wada
    • Journal Title

      Hearing Research 187

      Pages: 51-62

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Relationship between fluorescence intensity of GFP and the expression level of prestin in a prestin-expressing Chinese hamster ovary cell line2004

    • Author(s)
      Koji Iida, et al.
    • Journal Title

      JSME International Journal 47C

      Pages: 970-976

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Imaging of the cortical cytoskeleton of guinea pig outer hair cells using atomic force microscopy2004

    • Author(s)
      Hiroshi Wada, et al.
    • Journal Title

      Hearing Research 187

      Pages: 51-62

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Biomechanics at Micro- and Nanoscale Levels, Introduction of the Project.2004

    • Author(s)
      Hiroshi Wada
    • Journal Title

      Sanko Printing, Japan

      Pages: 3-6

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Stable expression of the motor protein in Chinese hamster ovary cells2003

    • Author(s)
      Koji Iida
    • Journal Title

      JSME International Journal 46C

      Pages: 1266-1274

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Stable expression of the motor protein prestin in Chinese hamster ovary cells2003

    • Author(s)
      Koji Iida, et al.
    • Journal Title

      JSME International Journal 46C

      Pages: 1266-1274

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Imaging by atomic force microscopy of the motor protein prestin using prestin-transfected CHO cells2006

    • Author(s)
      Hiroshi Wada
    • Organizer
      The 2nd Shanghai International Conference On Physiological Biophysics - audition & vision
    • Place of Presentation
      Shanghai, China.
    • Year and Date
      2006-11-06
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Imaging by atomic force microscopy of the motor protein prestin using prestin-transfected CHO cells.2006

    • Author(s)
      Hiroshi Wada
    • Organizer
      The 2nd Shanghai International Conference On Physiological Biophysics - audition & vision
    • Place of Presentation
      Shanghai, China.
    • Year and Date
      2006-11-06
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] 遺伝子導入と原子間力顕微鏡を用いたタンパク質モータプレスチンの可視化2006

    • Author(s)
      和田仁
    • Organizer
      第51回日本聴覚医学会総会・学術講演会
    • Place of Presentation
      山形, 日本
    • Year and Date
      2006-09-28
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Visualization of the motor protein prestin by gene introduction and atomic force microscopy.2006

    • Author(s)
      Hiroshi Wada
    • Organizer
      The 51st Congress of Japan Audiological Society
    • Place of Presentation
      Yamagata, Japan
    • Year and Date
      2006-09-28
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Topological characterization by atomic force microscopy of prestin2006

    • Author(s)
      Hiroshi Wada
    • Organizer
      The 5th World Congress of Biomechanics
    • Place of Presentation
      Munich, Germany.
    • Year and Date
      2006-07-31
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Topological characterization by atomic force microscopy of prestin.2006

    • Author(s)
      Hiroshi Wada
    • Organizer
      The 5th World Congress of Biomechanics
    • Place of Presentation
      Munich, Germany.
    • Year and Date
      2006-07-31
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] 電位依存性タンパク質モータprestinの構造変化観察, Atomic Force Microscopic Observation of the Conformational Changes in the Voltage-dependent Motor Protein Prestin2006

    • Author(s)
      和田仁
    • Organizer
      第18回バイオエンジニアリング講演会
    • Place of Presentation
      新潟, 日本
    • Year and Date
      2006-01-13
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Atomic Force Microscopic Observation of the Conformational Changes in the Voltage-dependent Motor Protein Prestin.2006

    • Author(s)
      Hiroshi Wada
    • Organizer
      The 18th Bioengineering Conference
    • Place of Presentation
      Niigata, Japan
    • Year and Date
      2006-01-13
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Atomic Force Microscopic Analysis of Prestin-transfected CHO Cell Plasma Membranes2005

    • Author(s)
      Michio Murakoshi
    • Organizer
      The 6th International Symposium on Future Medical Engineering Based on Bio-nanotechnology
    • Place of Presentation
      Sendai, Miyagi, Japan.
    • Year and Date
      2005-11-21
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Atomic Force Microscopic Analysis of Prestin-transfected CHO Cell Plasma Membranes.2005

    • Author(s)
      Michio Murakoshi
    • Organizer
      The 6th International Symposium on Future Medical Engineering Based on Bio-nanotechnology
    • Place of Presentation
      Sendai, Miyagi, Japan
    • Year and Date
      2005-11-21
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Atomic force microscopic imaging of the intracellular membrane surface of prestin-expressing Chinese hamster ovary cells2005

    • Author(s)
      Hiroshi Wada
    • Organizer
      The 9th “Mechanics of Hearing" Workshop
    • Place of Presentation
      Portland, Oregon, U. S. A.
    • Year and Date
      2005-07-24
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Atomic force microscopic imaging of the intracellular membrane surface of prestin-expressing Chinese hamster ovary cells.2005

    • Author(s)
      Hiroshi Wada
    • Organizer
      The 9th “Mechanics of Hearing" Workshop
    • Place of Presentation
      Portland, Oregon, U.S.A.
    • Year and Date
      2005-07-24
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Imaging of the cytoplasmic face of the prestin-expressing CHO cell membrane by atomic force microscopy.2005

    • Author(s)
      Michio Murakoshi
    • Organizer
      The 28th Annual Midwinter Research Meeting of the Association for Research in Otolaryngology
    • Place of Presentation
      New Orleans, Louisiana, U. S. A.
    • Year and Date
      2005-02-22
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Imaging of the cytoplasmic face of the prestin-expressing CHO cell membrane by atomic force microscopy.2005

    • Author(s)
      Michio Murakoshi
    • Organizer
      The 28th Annual MidWinter Research Meeting of the Association for Research in Otolaryngology
    • Place of Presentation
      New Orleans, Louisiana, U.S.A.
    • Year and Date
      2005-02-22
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] タンパク質モータプレスチン発現CHO細胞の原形質膜の観察-原子間力顕微鏡を用いて-, Observation of the plasma membrane of the CHO cells expressing the motor protein prestin by atomic force microscopy2005

    • Author(s)
      村越道生
    • Organizer
      第17回バイオエンジニアリング講演会
    • Place of Presentation
      名古屋, 愛知, 日本
    • Year and Date
      2005-01-22
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Presentation] Observation of the plasma membrane of the CHO cells expressing the motor protein prestin by atomic force microscopy.2005

    • Author(s)
      Michio Murakoshi
    • Organizer
      The 17th Bioengineering Conference
    • Place of Presentation
      Nagoya, Aichi, Japan.
    • Year and Date
      2005-01-22
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] Biomechanics at Micro- and Nanoscale levels, Vol. IV2007

    • Author(s)
      Hiroshi Wada
    • Publisher
      World Scientific Publishing
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] Medical Bio 組織・細胞の力学現象と制御2007

    • Author(s)
      幸野友浩
    • Publisher
      (株)オーム社
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] 生体医工学. 46(1)解説特集 : マイクロ・ナノバイオメカニクス2007

    • Author(s)
      安藤譲二
    • Publisher
      (社)日本生体医工学会
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] Biomechanics at Micro- and Nanoscale levels, Vol. II2006

    • Author(s)
      Hiroshi Wada
    • Publisher
      World Scientific Publishing
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] Biomechanics at Micro- and Nanoscale levels. Vol. I2005

    • Author(s)
      Hiroshi Wada
    • Publisher
      World Scientific Publishing
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] Biomechanics at Micro- and Nanoscale Levels, Volume I2005

    • Author(s)
      Hiroshi Wada
    • Total Pages
      173
    • Publisher
      World Scientific Publishing Co. Pte. Ltd.
    • Related Report
      2004 Annual Research Report
  • [Book] Biomechanics at Micro- and Nanoscale levels, Introduction of the Project2004

    • Author(s)
      Hiroshi Wada
    • Publisher
      Sanko Printing
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Remarks] 「研究成果報告書概要(和文)」より

    • URL

      http://www.wadalab.mech.tohoku.ac.jp/

    • Related Report
      2006 Final Research Report Summary
  • [Publications] Hiroshi Wada: "Imaging of the cortical cytoskeleton of guinea pig outer hair cells using atomic force microscopy"Hearing Research. 187. 51-62 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Koji Iida: "Stable expression of the motor protein in Chinese hamster ovary cells"JSME International Journal. 46C. 1266-1274 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2018-03-28  

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