Reduction of Glycosphingolipid Levels in Lipid Rafts Affects the Expression State and Function of Glycosylphosphatidylinositol-anchored Proteins but Does Not Impair Signal Transduction via the T Cell Receptor

Masakazu Nagafuku, Kazuya Kabayama, Daisuke Oka, Akiko Kato, Shizue Tani-Ichi, Yukiko Shimada, Yoshiko Ohno-Iwashita, Shou Yamasaki, Takashi Saito, Kazuya Iwabuchi, Toshiyuki Hamaoka, Jin Ichi Inokuchi, Atsushi Kosugi

    Research output: Contribution to journalArticle

    40 Citations (Scopus)

    Abstract

    Lipid rafts are highly enriched in cholesterol and sphingolipids. In contrast to many reports that verify the importance of cholesterol among raft lipid components, studies that address the role of sphingolipids in raft organization and function are scarce. Here, we investigate the role of glycosphingolipids (GSLs) in raft structure and raft-mediated signal transduction in T lymphocytes by the usage of a specific GSL synthesis inhibitor, D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP). Surface GM1 expression and the expression of GSLs in rafts were profoundly reduced by D-PDMP treatment, whereas the expression of other lipid and protein constituents, such as cholesterol, sphingomyelin, Lck, and linker for activation of T cells, was not affected. T cell receptor-mediated signal transduction induced by antigen stimulation or by antibody cross-linking was normal in D-PDMP-treated T cells. In contrast, the signal through glycosylphosphatidylinositol (GPI)-anchored proteins was clearly augmented by D-PDMP treatment. Moreover, GPI-anchored proteins became more susceptible to phosphatidylinositol-specific phospholipase C cleavage in D-PDMP-treated cells, demonsrating that GSL depletion from rafts primarily influences the expression state and function of GPI-anchored proteins. Finally, by comparing the effect of D-PDMP with that of methyl-β-cyclodextrin, we identified that compared with cholesterol depletion, GSL depletion has the opposite effect on the phosphatidylinositol-specific phospholipase C sensitivity and signaling ability of GPI-anchored proteins. These results indicate a specific role of GSLs in T cell membrane rafts that is dispensable for T cell receptor signaling but is important for the signal via GPI-anchored proteins.

    Original languageEnglish
    Pages (from-to)51920-51927
    Number of pages8
    JournalJournal of Biological Chemistry
    Volume278
    Issue number51
    DOIs
    Publication statusPublished - Dec 19 2003

    Fingerprint

    Glycosphingolipids
    Glycosylphosphatidylinositols
    Signal transduction
    T-Cell Antigen Receptor
    Signal Transduction
    T-cells
    Lipids
    Cholesterol
    Phosphoinositide Phospholipase C
    T-Lymphocytes
    Sphingolipids
    Proteins
    Sphingomyelins
    Cyclodextrins
    Cell membranes
    RV 538
    Chemical activation
    Cells
    Cell Membrane
    Antigens

    All Science Journal Classification (ASJC) codes

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Reduction of Glycosphingolipid Levels in Lipid Rafts Affects the Expression State and Function of Glycosylphosphatidylinositol-anchored Proteins but Does Not Impair Signal Transduction via the T Cell Receptor. / Nagafuku, Masakazu; Kabayama, Kazuya; Oka, Daisuke; Kato, Akiko; Tani-Ichi, Shizue; Shimada, Yukiko; Ohno-Iwashita, Yoshiko; Yamasaki, Shou; Saito, Takashi; Iwabuchi, Kazuya; Hamaoka, Toshiyuki; Inokuchi, Jin Ichi; Kosugi, Atsushi.

    In: Journal of Biological Chemistry, Vol. 278, No. 51, 19.12.2003, p. 51920-51927.

    Research output: Contribution to journalArticle

    Nagafuku, M, Kabayama, K, Oka, D, Kato, A, Tani-Ichi, S, Shimada, Y, Ohno-Iwashita, Y, Yamasaki, S, Saito, T, Iwabuchi, K, Hamaoka, T, Inokuchi, JI & Kosugi, A 2003, 'Reduction of Glycosphingolipid Levels in Lipid Rafts Affects the Expression State and Function of Glycosylphosphatidylinositol-anchored Proteins but Does Not Impair Signal Transduction via the T Cell Receptor', Journal of Biological Chemistry, vol. 278, no. 51, pp. 51920-51927. https://doi.org/10.1074/jbc.M307674200
    Nagafuku, Masakazu ; Kabayama, Kazuya ; Oka, Daisuke ; Kato, Akiko ; Tani-Ichi, Shizue ; Shimada, Yukiko ; Ohno-Iwashita, Yoshiko ; Yamasaki, Shou ; Saito, Takashi ; Iwabuchi, Kazuya ; Hamaoka, Toshiyuki ; Inokuchi, Jin Ichi ; Kosugi, Atsushi. / Reduction of Glycosphingolipid Levels in Lipid Rafts Affects the Expression State and Function of Glycosylphosphatidylinositol-anchored Proteins but Does Not Impair Signal Transduction via the T Cell Receptor. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 51. pp. 51920-51927.
    @article{e05b1743d1ad4342ab87607e458a62cc,
    title = "Reduction of Glycosphingolipid Levels in Lipid Rafts Affects the Expression State and Function of Glycosylphosphatidylinositol-anchored Proteins but Does Not Impair Signal Transduction via the T Cell Receptor",
    abstract = "Lipid rafts are highly enriched in cholesterol and sphingolipids. In contrast to many reports that verify the importance of cholesterol among raft lipid components, studies that address the role of sphingolipids in raft organization and function are scarce. Here, we investigate the role of glycosphingolipids (GSLs) in raft structure and raft-mediated signal transduction in T lymphocytes by the usage of a specific GSL synthesis inhibitor, D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP). Surface GM1 expression and the expression of GSLs in rafts were profoundly reduced by D-PDMP treatment, whereas the expression of other lipid and protein constituents, such as cholesterol, sphingomyelin, Lck, and linker for activation of T cells, was not affected. T cell receptor-mediated signal transduction induced by antigen stimulation or by antibody cross-linking was normal in D-PDMP-treated T cells. In contrast, the signal through glycosylphosphatidylinositol (GPI)-anchored proteins was clearly augmented by D-PDMP treatment. Moreover, GPI-anchored proteins became more susceptible to phosphatidylinositol-specific phospholipase C cleavage in D-PDMP-treated cells, demonsrating that GSL depletion from rafts primarily influences the expression state and function of GPI-anchored proteins. Finally, by comparing the effect of D-PDMP with that of methyl-β-cyclodextrin, we identified that compared with cholesterol depletion, GSL depletion has the opposite effect on the phosphatidylinositol-specific phospholipase C sensitivity and signaling ability of GPI-anchored proteins. These results indicate a specific role of GSLs in T cell membrane rafts that is dispensable for T cell receptor signaling but is important for the signal via GPI-anchored proteins.",
    author = "Masakazu Nagafuku and Kazuya Kabayama and Daisuke Oka and Akiko Kato and Shizue Tani-Ichi and Yukiko Shimada and Yoshiko Ohno-Iwashita and Shou Yamasaki and Takashi Saito and Kazuya Iwabuchi and Toshiyuki Hamaoka and Inokuchi, {Jin Ichi} and Atsushi Kosugi",
    year = "2003",
    month = "12",
    day = "19",
    doi = "10.1074/jbc.M307674200",
    language = "English",
    volume = "278",
    pages = "51920--51927",
    journal = "Journal of Biological Chemistry",
    issn = "0021-9258",
    publisher = "American Society for Biochemistry and Molecular Biology Inc.",
    number = "51",

    }

    TY - JOUR

    T1 - Reduction of Glycosphingolipid Levels in Lipid Rafts Affects the Expression State and Function of Glycosylphosphatidylinositol-anchored Proteins but Does Not Impair Signal Transduction via the T Cell Receptor

    AU - Nagafuku, Masakazu

    AU - Kabayama, Kazuya

    AU - Oka, Daisuke

    AU - Kato, Akiko

    AU - Tani-Ichi, Shizue

    AU - Shimada, Yukiko

    AU - Ohno-Iwashita, Yoshiko

    AU - Yamasaki, Shou

    AU - Saito, Takashi

    AU - Iwabuchi, Kazuya

    AU - Hamaoka, Toshiyuki

    AU - Inokuchi, Jin Ichi

    AU - Kosugi, Atsushi

    PY - 2003/12/19

    Y1 - 2003/12/19

    N2 - Lipid rafts are highly enriched in cholesterol and sphingolipids. In contrast to many reports that verify the importance of cholesterol among raft lipid components, studies that address the role of sphingolipids in raft organization and function are scarce. Here, we investigate the role of glycosphingolipids (GSLs) in raft structure and raft-mediated signal transduction in T lymphocytes by the usage of a specific GSL synthesis inhibitor, D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP). Surface GM1 expression and the expression of GSLs in rafts were profoundly reduced by D-PDMP treatment, whereas the expression of other lipid and protein constituents, such as cholesterol, sphingomyelin, Lck, and linker for activation of T cells, was not affected. T cell receptor-mediated signal transduction induced by antigen stimulation or by antibody cross-linking was normal in D-PDMP-treated T cells. In contrast, the signal through glycosylphosphatidylinositol (GPI)-anchored proteins was clearly augmented by D-PDMP treatment. Moreover, GPI-anchored proteins became more susceptible to phosphatidylinositol-specific phospholipase C cleavage in D-PDMP-treated cells, demonsrating that GSL depletion from rafts primarily influences the expression state and function of GPI-anchored proteins. Finally, by comparing the effect of D-PDMP with that of methyl-β-cyclodextrin, we identified that compared with cholesterol depletion, GSL depletion has the opposite effect on the phosphatidylinositol-specific phospholipase C sensitivity and signaling ability of GPI-anchored proteins. These results indicate a specific role of GSLs in T cell membrane rafts that is dispensable for T cell receptor signaling but is important for the signal via GPI-anchored proteins.

    AB - Lipid rafts are highly enriched in cholesterol and sphingolipids. In contrast to many reports that verify the importance of cholesterol among raft lipid components, studies that address the role of sphingolipids in raft organization and function are scarce. Here, we investigate the role of glycosphingolipids (GSLs) in raft structure and raft-mediated signal transduction in T lymphocytes by the usage of a specific GSL synthesis inhibitor, D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP). Surface GM1 expression and the expression of GSLs in rafts were profoundly reduced by D-PDMP treatment, whereas the expression of other lipid and protein constituents, such as cholesterol, sphingomyelin, Lck, and linker for activation of T cells, was not affected. T cell receptor-mediated signal transduction induced by antigen stimulation or by antibody cross-linking was normal in D-PDMP-treated T cells. In contrast, the signal through glycosylphosphatidylinositol (GPI)-anchored proteins was clearly augmented by D-PDMP treatment. Moreover, GPI-anchored proteins became more susceptible to phosphatidylinositol-specific phospholipase C cleavage in D-PDMP-treated cells, demonsrating that GSL depletion from rafts primarily influences the expression state and function of GPI-anchored proteins. Finally, by comparing the effect of D-PDMP with that of methyl-β-cyclodextrin, we identified that compared with cholesterol depletion, GSL depletion has the opposite effect on the phosphatidylinositol-specific phospholipase C sensitivity and signaling ability of GPI-anchored proteins. These results indicate a specific role of GSLs in T cell membrane rafts that is dispensable for T cell receptor signaling but is important for the signal via GPI-anchored proteins.

    UR - http://www.scopus.com/inward/record.url?scp=9144272911&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=9144272911&partnerID=8YFLogxK

    U2 - 10.1074/jbc.M307674200

    DO - 10.1074/jbc.M307674200

    M3 - Article

    C2 - 14506277

    AN - SCOPUS:9144272911

    VL - 278

    SP - 51920

    EP - 51927

    JO - Journal of Biological Chemistry

    JF - Journal of Biological Chemistry

    SN - 0021-9258

    IS - 51

    ER -