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1988 Fiscal Year Final Research Report Summary

Analytical Studies of Plasma Membrane Biogenesis in Transporting Epithelia

Research Project

Project/Area Number 61480127
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionHamamatsu University School of Medicine

Principal Investigator

FUJITA Michiya  Hamamatsu University School of Medicine Professor, 医学部, 教授 (60014031)

Co-Investigator(Kenkyū-buntansha) TAKESUE Yoshiki  Gifu University Faculty Faculty of Liberal Arts Associate Professor, 教養部, 助教授 (50023651)
UEZATO Tadayoshi  Hamamatsu University School of Medicine Professor, 医学部, 助手 (40115465)
OHTA Hidehiko  Hamamatsu University School of Medicine Professor, 医学部, 助教授 (00014192)
Project Period (FY) 1986 – 1988
KeywordsEnterocyte / Renal tubular epithelial cell / Glucose transporter / Na,K-ATPase / Trehalase / Cytochalasin B / Membrane protein / 蛍光抗体
Research Abstract

1.Microvullus membrane(MVM) and basolateral membrane (BLM) were prepared from small intestinal epithelial cells and were characterized biochemically and morphologically.
2.Glucose transport by MVM vesicles was inhibited cytochalasin B (CYB). 52K protein was photolabeled by CYB. Glucose transport by MVM vesicles was inhibited maximally 60% by CYB (Ki=10 M) and 86K protein was labeled with CYB (Kd=8 M,Bmax=70pmol/mg protein).
3.Fluorescent antibody against 70K MVM protein reacted not only with intestinal MVM but with renal and hepatic MVM. Antibody against Na,K-ATPasereacted with intestinal, renal and hepatic BLM.
4.Trehalase of renal MVM turned out to be covalently bound with MVM viaphosphatidyl inositol. Trehalasesolubilized with Triton X-100 was hydrophobic and adsorbed to phenyl sepharose clumn. Iucubated at 37 c, it became hydrophilic and this conversion was not depressed by any protease inhibitors so far known.
6.Bioassembly of MVM and BLM proteins showed monophasic time course for MVM and biphasic one for BLM. Possible contamination of BLM with endoplasmic reticulum membrane is under investigation to account for discrepancy.

  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] 藤田道也: 膜. 12. 64-71 (1987)

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      「研究成果報告書概要(和文)」より
  • [Publications] 藤田道也: 生体の科学. 37. 365-366 (1986)

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  • [Publications] 藤田道也: 生体の科学. 38. 89-91 (1987)

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      「研究成果報告書概要(和文)」より
  • [Publications] UEZATO,T.: Biochimie. 70. 1775-1779 (1989)

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  • [Publications] Uezato,T.: Biochem.Int.12. 199-206 (1986)

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  • [Publications] Uezato,T.: J.Cell Science. 85. 177-185 (1986)

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  • [Publications] 太田英彦: 生体の科学. 38. 369-370 (1987)

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  • [Publications] 太田英彦: 生体の科学. 38. 416-417 (1987)

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  • [Publications] SUGIYAMA,Y.: HEPATOLOGY.

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  • [Publications] YOKOTA,K.: Biochim.Biophys.Acta. 881. 405-414 (1986)

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  • [Publications] TAKESUE,Y.: FEBS LETT.201. 5-8 (1986)

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  • [Publications] YOKOTA,K.: J.Biochem.103. 132-136 (1988)

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  • [Publications] 星猛: "新生理科学大系第18巻消化と吸収の生理学" 医学書院, 1-404 (1988)

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  • [Publications] Fujita, M.: "Intracellular mechanism of cell membrane domainization Membrane" Membrane(Maku). 12. 64-71 (1987)

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      「研究成果報告書概要(欧文)」より
  • [Publications] Fujita, M.: "Preparation of small intestinal brush borders" Seitai No Kagaku. 37. 365-366 (1986)

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      「研究成果報告書概要(欧文)」より
  • [Publications] Fujita, M.: "Disordered intracellular traffic and autoimmunization" Seitai No Kagaku. 38. 89-91 (1987)

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  • [Publications] Uezato, T.: "Cytochalasin B-sensitive, sodium ion-dependent glucose transport in intestinal microvillus membrane" Biochimie. 70. 1775-1779 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Uezato, T.: "Photoaffinity labelling of the K562 cell membrane D-glucose transporter with cytochalasin B" Biochem. Int.12. 199-206 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Uezato, T.: "Cytochalasin B-sensitive proteins related to glucose transport across the intestinal epithelial cell" J. Cell Sci.85. 177-185 (1986)

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      「研究成果報告書概要(欧文)」より
  • [Publications] Ohta, H.: "Gap junction protein" Seitai No Kagaku. 38. 369-370 (1987)

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  • [Publications] Ohta, H.: "Fluorescent labeling" Seitai No Kagaku. 38. 416-417 (1987)

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  • [Publications] Sugiyama, Y.: "Changes in density and distribution of gap junctions after partial hepatectomy: Immunochemical and morphometric studies" Hepatology.

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  • [Publications] Yokota, K.: "Purification and characterization of amphiphilic trehalase from rabbit small intestine" Biochim. Biophys. Acta. 881. 405-414 (1986)

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  • [Publications] Takesue, Y.: "Solubilization of trehalase from rabbit renal and intestinal brush-border membranes by a phosphatidyl inositol-specific phospholipase C" FEBS LETT.201. 5-8 (1986)

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  • [Publications] Yokota, K.: "In vitro loss of hydrophobicity of trehalase from the brush border membrane of rabbit kidney cortex" J. Biochem.103. 132-136 (1988)

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  • [Publications] Hoshi, T.: Digestion and Absorption (Handbook of Physiology: New Series Vol.18). Igaku-Shoin Ltd., 404 (1988)

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Published: 1990-03-20  

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