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ISOLATION OF ION CHANNEL MOLECULES BY USING LIPID BILAYER TECHNIQUE AS AN ASSAYING SYSTEM AND STUDY ON THEIR STRUCTURE AND FUNCTION RELATIONSHIP

Research Project

Project/Area Number 04454620
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 生物物性学
Research InstitutionOsaka University

Principal Investigator

KASAI Michiki  OSAKA Univ., FAC.ENGINEER.SCI., PROFESSOR, 基礎工学部, 教授 (40022595)

Co-Investigator(Kenkyū-buntansha) IDE Toru  OSAKA Univ., FAC.ENGINEER.SCI., ASSISTANT, 基礎工学部, 助手 (60231148)
TAGUCHI Takahisa  OSAKA Univ., FAC.ENGINEER.SCI., ASSISTANT, 基礎工学部, 助手 (10197246)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1992: ¥4,200,000 (Direct Cost: ¥4,200,000)
KeywordsIon channel / Lipid bilayr / K channel / CI channel / Sarcoplasmic reticulum / Ca channel
Research Abstract

Purification of ion channels in sacrcoplasmic (SR) of rabbit skeletal muscle has been carried out by assaying the channel activity in the lipid bilayr system. As a result, the fraction which contains 138 and 100 kDa protein showed CI^- channel activity. After purification of 138 kDa protein, partial amino acid sequences were determined and their homegogy was referred. The 138 kDa protein was found to be the same one as that reported as 160 kDa protein which has Ca^<2+>-binding activity. The difference was due to the nuderestimation of the previous workers. The 138 kDa protein interacts with 100 kDa protein in SR.When 100 kDa protein was incorporated into lipid bilayr, it showed channel activity. It was suggested that 138 and 100 kDa proteins worked as ion channle by making complex.Antibody against 138 kDa protein inhibited slow KCI permaetion through SR vesicles.
CI^- channel activity in transverse tubule (T-tubule) in rabbit skeletal muscle was investigated by lipid bilayr system. A back-ground type CI^- channel (open at resting potential) having 40 pS was found. This channel was blocked by 9-AC,DIDS,SITS,and EA which were known as blockers againg voltage dependent anion channels. Further, the channel was blocked by IAA-94 which was developed as a blocker for CI^- channel in apical membrane from trachea. Similarity between these channels was suggested. Antibody which blocked the channel activity recognized 90 kDa protein in muscle.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] T.Taguchi: "Immunological identification of a Cl^--channel protein in electric organs of Narke japonica" Biochem.Biophys.Res.Commun.188. 1228-1234 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Kasai: "Channel opening process is responsible for the slow component in the flux measurement of membrane vesicles." J.Theor.Biol.161. 461-480 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Kasai: "Effects of ryanodion on permeability of choline and glucose through calcium channel in sarcoplasmic reticulum vesicles." J.Biochem.113. 327-333 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] J.Hidaka: "Characterizaion of a Cl^--channel from rabbit transverse tubules in the planar lipid bilayer system." Biochem.Biophys.Res.Commun.191. 977-982 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] J.Hidaka: "An anion channel of transverse tubules incorporated into planar lipid bilayers:Single channel behavior and conductance properties" Ann.New York Acad.Sci.707. 424-426 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Kawasaki: "Regulation of Calcium Channel in Sarcoplasmic Reticulum by Calsequestrin" Biochem.Biophys.Res.commun.199. 1120-1127 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Kasai: "Effects of KC1 concentrations on gating properties of calcium release channels in sarcoplasmic reticulum vesiclis" J.Biochem. 117. 251-256 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Ide: "An anion channel from transverse tubular membranes incorporated into planar lipid bilayers" Biochim.Biophys.Acta. (印刷中). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Taguchi.: "Immunological identification of a CI^--channel protein in electric organs of Narke japonica" Biochem.Biophys.Res.Commun.188. 1228-1234 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Kasai.: "Channel opening process is responsible for the slow component in the flux measurement of membrane vesicles." J.Theor.Biol.161. 461-480 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Kasai.: "Effects of ryanodine on permeability of choline and glucose through calcium channel in sarcoplasmic reticulum vesicles." J.Biochem.113. 327-333 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] J.Hidaka.: "Characterization of a CI^--channel from rabbit transverse tubules in the planar lipid bilayr system." Biochem.Biophys.Res.Commun.191. 977-982 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] J.Hidaka.: "An anion channel of transverse tubules incorpotated into planar lipid bilayrs : Single channel behavior and conductance properties" Ann.New York Acad.Sci.707. 424-426 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Kawasaki.: "Regulation of Calcium Channel in Sarcoplasmic Reticulum by Calsequestrin" Biochem.Biophys.Res.Commun.199. 1120-1127 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Kasai.: "Effects of KCI concentrations on gating properties of calcium release channels in sarcoplasmic reticulum vesicles" J.Biochem.117. 251-256 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Ide.: "An anion channel of transverse tubular membranes incorporated into planar lipid bilayrs" Biochem.Biophys.Acta. (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Kawasaki: "Regulation of Calcium Channel in Sarcoplasmic Reticulum by Calsequestrin" Biochem.Biophys.Res.Commun.199. 1120-1127 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] X.-X.Bo: "Synapse formation in vitro under glia-free condition" Neurscience Research. s19. s45- (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Kasai: "Effects of KCl concentrations on gating properties of calcium release channels in sarcoplasmic reticulum vesicles" J.Biochem.117. 251-256 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Ide: "An anion channel from transverse tubular membranes incorporated into planar lipid bilayers" Biochim.Biophys.Acta. (印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 葛西道夫: "光散乱法.膜学実験シリーズ 1." 共立出版, 228-291 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 葛西道夫: "電気穿孔法,物理薬化学実験講座 2巻." 広川書店(印刷中), (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Kasai: "Channel opening process is responsible for the slow component in the flux measurement of membrane vesicles." J.Theor.Biol.161. 461-480 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Kasai: "Effects of ryanodine on permeability of choline and glucose through calcium channel in sarcoplasmic reticulum vesicles." J.Biochem.113. 327-333 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] J.Hidaka: "Characterization of a Cl-channel from rabbit transverse tubules in the planar lipid bilayer system." Biochem.Biophys.Res.Commun.191. 977-982 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Sato: "Making synaptic vesicles fuse with lipid bilayers.Reply." Biophys.J.65. 975-975 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] J.Hidaka: "An anion channel of transverse tubules incorporated into planar lipid bilayers:Single channel behavior and conductance properties" Ann.New York Acad.Sci.(印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Taguchi,T.,Kawasaki,T.Kasai,M.: "Immunological identification of a Cl^-channel protein in electric organs of Narke japonica." Biochem.Biophys.Res.Comm.188. 1228-1234 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hidaka,J.Ide,T.Taguchi,T.Kasai,M.: "Identification of an anion channel protein from transverse tubules of rabbit skeletel mnscle." The annals of the New York acsdemy of Science. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hidaka,J.Ide,T Taguchi,T.Kasai,M.: "Charecterization of a Cl^--channel from rabbit transverse tubules in the planar lipid bilayer system." Biochem.Biophys.Res.Comm.(1993)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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