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Analysis of gastric acid secretary mechanism by identifying proton pomp beta subunit binding protein

Research Project

Project/Area Number 12670521
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Gastroenterology
Research InstitutionTokyo Woman's Medical University

Principal Investigator

SAITOH Toshihito  Tokyo Women's Medical University Dept. of Internal Medicine Associate Professor, 医学部, 講師 (50246609)

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)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
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)
Keywordsproton pump H+K+ATPase / GSTfusion protein / gastric acid secretion / βサブユニット
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.

Report

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

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Otsuka H, et al.: "Long-lasting binding of IT-066 to human histamine H_2 receptor"Digestive Diseases & Science. 45. 796-801 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Fukushima Y. et al.: "Palmitoypation of the carmine histamine H_2 receptor occurs at Gys(305)"Biochimica et Biophysica Acta. 1539. 181-191 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Fukushima Y. et al.: "G649, an alleic variant of the human H_2 receptor with low basal"The pharmacogenomic Journal. 1. 78-83 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Fukushima Y. et al.: "Potent and longactim of lafutidine on the human histamine"Digestion. 64. 155-160 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Saitoh T. et al.: "Effects of N-alpha-methyl-histamine on human H_2 receptor"Gut. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Saitoh T. et al.: "Intragustric acidity and circadian rhythm"Biomed. Pharmacothr. 55. 138-141 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Otsuka H, Fukushima Y, Tamai M, Takahashi H, Mori H, Asano T, Katsube T, Ogawa K, Kajiwara T, Ohkawa SI Saitoh T,: "Long-lasting binding of IT-066 to human histamine H2 receptor."Digestive Diseases & Science. 45. 796-801 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Saitoh T, Otsuka K, Ohkawa S.I: "Intragastric pH and circadiun rhythm."Asian Medical Journal. 43 (8). 353-60 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Fukushima Y, Saitoh T, Anai M, Ogihara T, Inukai K, Funaki M, Sakoda H, Onishi Y, Ono H, Fujishiro M, Ishikawa T, Takata K, Nagai R, Omata M, Asano T: "Palmitoylation of the canine histamine H2 receptor occurs at Cys(305) and is important for cell surface targeting."Biochimica et Biophysica Acta. 1539. 181-91 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Saitoh T, Watanebe Y, Kubo Y, Shinagawa M, Otsuka K, Ohkawa S.I, Watanabe T,: "Intragastric acidity and circadiun rhythm. Biomed."Pharmacothr. 55. 138-41 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Fukushima Y, Saitoh T, Anai M, et al.: "G649, an alleic variant of the human H2 receptor with low basal activity, is redidtant to upregulation upon antagonist exporsure."The Pharmacogenomics Journal. 1. 78-83 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Fukushima Y, Otsuka H, Ishikawa M, Saitoh T.: "Potent and long action of lafutidine on the human histamine H2 receptor."Digestion. 64. 155-60 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Saitoh T, Fukushima Y, Otsuka H, et al.: "Effects of N-alpha-methyl-histamine on human H2 receptors expressed in CHO cells."Gut (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Otsuka H. et al.: "Long-lasting binding of IT-066 to human histamine H_2 receptor"Digestive Diseases & Sciences. 45. 796-801 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Fukushima Y. et al.: "Palmitoylation of the canine histamine H_2 receptor occurs at Cys(305) and is important"Biochimica et Biophysica Acta. 1539. 181-191 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Fukushima Y. et al.: "G649, an alleic variant of the human H_2 receptor with low basal"The Pharmacogenomics Journal. 1. 78-83 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Fukushima Y. et al.: "Potent and long action of lafutidine on the human histamine"Digestion. 64. 155-160 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Saitoh T. et al.: "Effects of N-alpha-methyl-histamine on human H_2 receptors"Gut. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] Saitoh T.: "Intragastric acidity and circadian rhythm"Biomed. Pharmacothr. 55. 138-141 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 斉藤寿仁: "プロトンポンプαサブユニットN端のリン酸化についての検討"Therapeutic Research. 22. 12-16 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Otsuka Hiroko: "Long-lasting binding of IT-066 to the human histamine H_2 receptor"Digestive Diseases and Sciences. 45. 186-192 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 斉藤寿仁: "各種PPIの投与初期における胃内pHの変動"胃分泌研究会誌. 32. 9-12 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] SAITOH Toshihito: "INTRAGASTRIC PH AND CIRCADIAN RHYTHM"ASIAN MEDICAL JOURNAL. 43・8. 353-360 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Saitoh Toshihito: "Effects of Nalpha-methyl-histamine on human H_2 receptor"Gut. 93(印刷中). (2001)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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