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A new functional analysis of intracellular ion-transporting proteins

Research Project

Project/Area Number 24659114
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeSingle-year Grants
Research Field General pharmacology
Research InstitutionKyoto University

Principal Investigator

KANEKO Shuji  京都大学, 薬学研究科(研究院), 教授 (60177516)

Co-Investigator(Kenkyū-buntansha) NAKAGAWA Takayuki  京都大学, 大学院薬学研究科, 准教授 (30303845)
Co-Investigator(Renkei-kenkyūsha) SHIRAKAWA Hisashi  京都大学, 大学院薬学研究科, 助教 (50402798)
Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsイオン輸送 / トランスポータ / 細胞内小器官 / シナプス小胞 / 自己組織化単分子層 / 電気生理学
Research Abstract

A novel electrophysiological assay technique based on solid supported membranes (SSM) has been shown to be useful for analysis of channels and transporters. We tried to apply this technique to synaptic vesicles (SVs) and lysosomal membranes (LMs), purified from rat brains and mouse livers, respectively. Application of ATP to the sensors with SVs triggered current responses dependent on Mg2+ and sensitive to bafilomycin A1 (Baf), which are thought to be V-ATPase specific currents. On the other hand, though LMs are also embedded with V-ATPases, the sensors evoked current responses with different characteristics. They were independent of Mg2+, only partially sensitive to Baf and even triggered by application of ADP. These features suggest that the currents are not attributed to V-ATPases but any other lysosomal protein. The present study shows the utility and potential of SSM-based electrophysiology especially for analyzing native organellar membrane proteins.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (3 results)

All 2013 2012 Other

All Journal Article (1 results) Presentation (1 results) Remarks (1 results)

  • [Journal Article] Application of a novel electrophysiological assay technique to native lysosomal membranes2013

    • Author(s)
      Yu Sakamoto
    • Journal Title

      京都大学薬学部卒業研究論文集

      Volume: 11巻1-6頁

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
  • [Presentation] トランスポータ活性を電気的に測定する自動化システム2012

    • Author(s)
      金子周司
    • Organizer
      第34回生体膜と薬物の相互作用シンポジウム
    • Place of Presentation
      京都
    • Related Report
      2013 Final Research Report
  • [Remarks]

    • URL

      http://www.pharm.kyoto-u.ac.jp//channel/ja/

    • Related Report
      2013 Final Research Report

URL: 

Published: 2013-05-31   Modified: 2019-07-29  

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