| Project/Area Number |
09680664
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Okazaki National Research Institutes |
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Principal Investigator |
OIKI Shigetoshi National Institute for Physiological Sciences, Department of Cellular and Molecular Physiology, Okazaki National Research Institutes, Associate Professor, 生理学研究所, 助教授 (10185176)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | inward rectifier / hyperpolarization / surface potential / open channel block / kinetics / ブロック / 単一チャネル記録 / 電位依存性 / ゲーティング / ブロッカー |
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| Research Abstract |
In 1998 the crystal structure of the potassium channel was elucidated by MacKinnon's group and the structural parts of the gate was directly demonstrated. Based on this structural information some mutations were introduced into two kinds of potassium channels (Kir2.1, KAT1). For the inwardly rectifying potassium channels (Kir2.1) three charged residues exist in the loop between the pore helix and the inner helix. The negatively charged residues (AspィイD1152ィエD1, GluィイD1153ィエD1) were neutralized. We developed a method to evaluate the surface potential from the shift of the chord conductance-voltage curves. It was found that GluィイD1153ィエD1 modified the affinity of MgィイD12+ィエD1 ion and reduced the surface potential, thus decreasing the single channel conductance. When the concentration of external MgィイD12+ィエD1 was increased the open channel blocking appeared. From the kinetic analyses of the single channel recordings under MgィイD12+ィエD1 blocking condition it was suggested that MgィイD12+ィエD1 ion in the blocked state inhibited the closure of the gate. In the KAT1 channel a mutation in the C-terminal end of the pore helix (T256) altered the activation kinetics upon hyperpolarization. It was revealed that the transition rate between the closed state and the open state was susceptible to the mutation.
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