| Project/Area Number |
02304067
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| Research Category |
Grant-in-Aid for Co-operative Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
分子遺伝学・分子生理学
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| Research Institution | Himeji Institute of Technology, Faculty of Science (1991)
Jichi Medical University (1990) |
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Principal Investigator |
HIRATA Hajime Himeji Inst. Tech., Faculty Sci., Prof., 理学部, 教授 (40049052)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAGI Masayuki Osaka Univ., Educ. Sch., Assoc. Prof., 教養部, 助教授 (20028145)
SOKABE Masahiro Nagoya Univ., Med. Sch., Assoc. Prof., 医学部, 助教授 (10093428)
TSUDA Motoyuki Himeji Inst. Tech., Faculty Sci., Prof., 理学部, 教授 (60045458)
KIRINO Yutaka Kyushu Univ., Faculty Pharm., Prof., 薬学部, 教授 (10012668)
KASAI Michiki Osaka Univ., Faculty Eng. Sci., Prof., 基礎工学部, 教授 (40022595)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 1991: ¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1990: ¥6,300,000 (Direct Cost: ¥6,300,000)
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| Keywords | Phospholipid Planar Bilayer / Ion Channel / Ion Pump / Signal Transduction / Functional Membrane Protein / Synthetic Peptide / Cryo-Electron Microscopy / Computer Simulation / イオンチャンネル |
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| Research Abstract |
1) The Cl^--channels in synaptic vesicle membranes were characterized after reconstituting into planar bilayers and classified into two types of activities. It was also demonstrated that fusion of synaptic vesicles with liposomes was triggered by soiie unknown protein factor (s). 2) A synthetic polyether in combination with quaternary amine compound formed a K^+ specific ion channel in planar bilayers. Chemical modification of gramicidin as well as synthetic peptides was introduced for characterization of ion permeation through ion channel molecules. 3) Amphiphilic peptides composed of 20 amino acids from N-terminal segment of influenza virus were synthesized and their membrane fusion acitvities were characterized. 4) Using planar bilayers as an assay procedure, K^+-channel and Cl^--channel proteins were purified from sarcoplasmic reticulum membranes. 5) Ion channel activities in giant liposomes prepared by various methods were characterized in detail comparing with those in planar bilayers. 6) A new method, a pinching oil layer method, for reconstituting ion channel proteins was developed. 7) Detailed analyses on interaction between rhodopsin and G-protein in octopus photoreceptor cells was carried out. And a cGMP activated cation channel in octopus photoreceptor cells was identified using planar bilayers. 8) Reconstitution of either bacteriorhodopsin or H^+-ATPase into planar bilayers was successfully carried out by newly developed methods ; a direct insertion method or membrane fusion peptide method. 9) Electron microscopic studies on Cl^--channel of frog muscle spindle were carried out and its structure was deduced. 10) A new time resolution cryoelectron microscope c technique was developed for studies of conformational changes of proteins at time resolution of 5 ms. 11) Visualization of diffusion of phospholipid molecules in phospholipid bilayer membranes was performed by computer simulation, and permeation of oxygen molecules were analyzed.
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