| Project/Area Number |
12670521
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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 |
Gastroenterology
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| Research Institution | Tokyo Woman's Medical University |
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Principal Investigator |
SAITOH Toshihito Tokyo Women's Medical University Dept. of Internal Medicine Associate Professor, 医学部, 講師 (50246609)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUSHIMA Yasushi Univ. of Tokyo, Faculty of Medicine, Assistant Professor, 医学部, 医員
TAKAHASHI Haruki Tokyo Women' s Medical Univ. Dept. of Internal Medicine Assistant, 医学部, 助手 (00246612)
OTUKA Hiroko Tokyo Women' s Medical Univ. Dept. of Internal Medicine Assistant Professor, 医学部, 助手 (20307606)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | proton pump H+K+ATPase / GSTfusion protein / gastric acid secretion / βサブユニット |
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| Research Abstract |
We made peptides constituted about 30 amino acids, which exist intracellular portion of N end of proton pump H+K+ATPase beta subunit. Applying this peptides to the affinylbeads, the column was prepared for the purpose of identification of unknown proteins that bind to those peptides. Rat gastric mucosa lysate was applied to the column and the extraction of unknown proteins was done, followed by electrophoresis. Amino acid sequencing of the isolated protein was done after electrophoresis. The localization of this protein in the parietal cells in gastric mucosa was studied by immunohistochemical method. Isotope labeled GST-intracellular beta subunit portion fusion protein as the probe for expression cloning was prepared. It has been suggested that there are proteins which regulate proton pump function with or without the stimuli of gastric acid secretion. Using the affinity column system and expression cloning conducting by GST fusion protein, we could isolate the candidate protein that binds to beta submit of proton pump H+K+ATPase and clarify the interaction between the binding proteins and beta subunit.
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