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2005 Fiscal Year Final Research Report Summary

Structure and function of cholesterol-enriched membrane microdomains (lipid rafts)

Research Project

Project/Area Number 16570127
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research InstitutionTokyo Metropolitan Foundation for Research on Aging and Promotion of Human Welfare

Principal Investigator

IWASHITA Yoshiko  Tokyo Metropolitan Foundation for Research on Aging and Promotion of Human Welfare, Tokyo Metropolitan Institute of Gerontology, Senior Research Scientist, 東京都老人総合研究所, 研究副部長 (50111498)

Project Period (FY) 2004 – 2005
KeywordsCHOLESTEROL / LIPID RAFTS / T LYMPHOCYTES / SIGNALING / PERRINGOLYSIN O
Research Abstract

We prepared a non-cytolytic derivative of perfringolysin O (θ-toxin), a cholesterol-binding cytolysin, as a probe for cholesterol, and demonstrated that the probe (BCθ) binds selectively to cholesterol in cholesterol-rich membrane domains of intact cells. Based on the finding that BCθ binds to a subpopulation of membranes among those recovered in detergent-insoluble, low-density fractions (raft fractions),' we isolated BCθ-bound membranes from the raft fractions of Jurkat T cells. The BCθ-bound membrane subpopulation has a much higher cholesterol/phospholipid (C/P) molar ratio (〜1.0) than the BCθ-unbound population in raft fractions (〜0.3). It contains not only the raft markers GM1 and flotillin, but also some T-cell receptor (TCR) signaling molecules, including Lck, Fyn and LAT. In addition, Csk and PAG, inhibitory molecules of the TCR signaling cascade, are also contained in the BCθ-bound membranes. On the other hand, CD3 and Zap70 are localized in the BCθ-unbound membranes, segregated from other TCR signaling molecules under non-stimulated conditions. However, upon stimulation of TCR, portions of CD3 and Zap70 are recruited to the BCθ-bound membranes. The Triton X-100 concentration used for lipid raft preparation affects neither the C/P ratio nor protein composition of the BCθ-bound membranes. These results show that our method is useful for isolating a particular cholesterol-rich membrane domain of T cells, which could be a core domain controlling the TCR signaling cascade.

  • Research Products

    (12 results)

All 2005 2004

All Journal Article (12 results)

  • [Journal Article] Separation of a cholesterol-enrichied microdomain involved in T-cell signal transduction2005

    • Author(s)
      Shimada, Y. et al.
    • Journal Title

      FEBS J. 272・21

      Pages: 5454-5463

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Role for upregulated ganglioside biosynthesis and association of Src family kidases with microdomains in retinoic acid-induced differentiation of F9 embryonal carcinoa cells2005

    • Author(s)
      Sato, T. et al.
    • Journal Title

      Glycobiology 15・7

      Pages: 687-699

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Adhesive surface determines raft composition in platelets adhered under flow2005

    • Author(s)
      van Lier, M. et al.
    • Journal Title

      J. Thromb. Haemost. 3・11

      Pages: 2514-2525

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Structure and function of liquid rafts in human activated T cells2005

    • Author(s)
      Tani-ichi, S. et al.
    • Journal Title

      International Immunology 17・6

      Pages: 749-758

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Distinct localization of liquid rafts and externalized phosphatidylserine at the surface of apoptotic cells2005

    • Author(s)
      Ishii, H. et al.
    • Journal Title

      Biochem, Biophys. Res. Commun. 327・1

      Pages: 94-99

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Separation of a cholesterol-enriched microdomain involved in T-cell signal transduction2005

    • Author(s)
      Shimada, Y. et al.
    • Journal Title

      FEBS J. 272(21)

      Pages: 5454-5463

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Role for upregulated ganglioside biosynthesis and association of Src family kinases with microdomains in retinoic acid-induced differentiation of F9 embryonal carcinoma cells2005

    • Author(s)
      Sato, T. et al.
    • Journal Title

      Glycobiology 15(7)

      Pages: 687-699

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Adhesive surface determines raft composition in platelets adhered under flow2005

    • Author(s)
      van Lier, M. et al.
    • Journal Title

      J.Thromb.Haemost. 3(11)

      Pages: 2514-2525

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Structure and function of lipid rafts in human activated T cells2005

    • Author(s)
      Tani-ichi, S. et al.
    • Journal Title

      International Immunology 17(6)

      Pages: 749-758

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Distinct localization of lipid rafts and externalized phosphatidylserine at the surface of apoptotic cells2005

    • Author(s)
      Ishii, H., et al.
    • Journal Title

      Biochem.Biophys.Res.Commun. 327(1)

      Pages: 94-99

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] コレステロールを見る2004

    • Author(s)
      嶋田有紀子, 岩下淑子
    • Journal Title

      蛋白質・核酸・酵素 49・10

      Pages: 1381-1387

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] How to visualize cholesterol2004

    • Author(s)
      Shimada, Y., Iwashita, Y.
    • Journal Title

      Protein, Nucleic Acid and Enzyme 49(10)

      Pages: 1381-1387

    • Description
      「研究成果報告書概要(欧文)」より

URL: 

Published: 2007-12-13  

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