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Voltage-dependent gating and block by internal spermine in the inwardly rectifying K channels

Research Project

Project/Area Number 11470013
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionKansai Medical University

Principal Investigator

MATSUDA Hiroko  Kansai Medical University, Faculty of Medicine, Professor, 医学部, 教授 (10181736)

Co-Investigator(Kenkyū-buntansha) OMORI Koicihro  Kansai Medical University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (80094465)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥11,300,000 (Direct Cost: ¥11,300,000)
Fiscal Year 2001: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2000: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1999: ¥5,700,000 (Direct Cost: ¥5,700,000)
KeywordsK channel / Inward rectification / spermine
Research Abstract

To investigate the mechanism of inward rectification, single-channel currents through inwardly rectifying K^+ (IRK1 ; Kir 2.1) channels were studied. cDNA encoding a wild-type (WT) IRK1 channel, D172N and tandem tetramer WT-(D172N)2-WT was transfected into COS-1 cells using the liposome method, and voltage clamp experiments were done after 48-72 h. Steady-state outward currents were recorded and open probability was calculated. The activation curve was fitted with single Boltzmann equation. The voltage of half-activation was 33.4 mV (WT), 51.2 mV (WT-(D172N)2-WT) and 76.6 mV (D172N). Open-and zero-current tirne histograms were constructed on current records at +42 mV The open-time histogram was fitted with a single exponential function. The mean open time was 53.1 ms (WT), 46.2 ms (WT-(D172N)2-WT) and 39.4 ms (D172N) in the control. Internal spermine reduced the open time in a concentration dependent manner: 37.8 ms (1 nM) and 11.7 ms (10 nM) in WT channels ; 34.7 ms (1 nM), 10.9 ms (10 nM) and 1.6 ms (100 nM) in WT-(D172N)2-WT channels ; 31.6 ms (1 nM), 11.7 ms (10 nM) and 1.7 ms (100 nM) in D172N channels. More than two exponential functions were necessary to fit the zero-cunent time histogram except in D172N channels with 10 and 100 nM spermine. These results suggest that intrinsic gating does exist separately from the spermine block. On the assumption of the state model (blocked state --- Open state --- closed state 1 --- closed state 2), blocking rates were estimatcd in each channel. They were 6.2-7.7 s^<-1> (1 nM), 64.7-70.1 s^<-1> (10 nM) and 563-603 s^<-1> (100 nM) and did not depend on the number of D172N mutant.

Report

(4 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (5 results)

All Other

All Publications (5 results)

  • [Publications] Matsuda H, Oishi K, Omori K: "Kinetics of the block of IRK1 and D172N mutant channels by internal ---"Japanese Journal of Physiology (Suppl). 49. S104 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Matsuda H., Oishi K and Omori K: "Kinetics of the block of IRK1 and D172N mutant hcannels by internal spermine"Japanese Journal of Physiology. 49(suppl). S104 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Matsuda H, Oishi K, Omori K: "Kinetics of the block of IRK1 and D172N mutant channels by internal"Japanese Journal of Physiology (Suppl). 49. S104 (1999)

    • Related Report
      2001 Annual Research Report
  • [Publications] Matsuda H,Oishi K & Omori K: "Kinetics of the block of IRK1 and D172N mutant channels by internal…"Japanese Journal of Physiology (Suppl). 49. S104 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Matsuda H,Oishi K & Omori K: "Kinetics of the block of IRK1 and D172N mutant channels by internal---"Japanese Journal of Physiology(Suppl). (印刷中).

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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