Identification of proteins associating with glycosylphosphatidylinositol- anchored T-cadherin on the surface of vascular endothelial cells

Role for Grp78/BiP in T-cadherin-dependent cell survival

Maria Philippova, Danila Ivanov, Manjunath B. Joshi, Emmanouil Kyriakakis, Katharina Rupp, Taras Afonyushkin, Valery Bochkov, Paul Erne, Therese J. Resink

    Research output: Contribution to journalArticle

    80 Citations (Scopus)

    Abstract

    There is scant knowledge regarding how cell surface lipid-anchored T-cadherin (T-cad) transmits signals through the plasma membrane to its intracellular targets. This study aimed to identify membrane proteins colocalizing with atypical glycosylphosphatidylinositol (GPI)-anchored T-cad on the surface of endothelial cells and to evaluate their role as signaling adaptors for T-cad. Application of coimmunoprecipitation from endothelial cells expressing c-myc-tagged T-cad and high-performance liquid chromatography revealed putative association of T-cad with the following proteins: glucose-related protein GRP78, GABA-A receptor α1 subunit, integrin β3, and two hypothetical proteins, LOC124245 and FLJ32070. Association of Grp78 and integrin β3 with T-cad on the cell surface was confirmed by surface biotinylation and reciprocal immunoprecipitation and by confocal microscopy. Use of anti-Grp78 blocking antibodies, Grp78 small interfering RNA, and coexpression of constitutively active Akt demonstrated an essential role for surface Grp78 in T-cad-dependent survival signal transduction via Akt in endothelial cells. The findings herein are relevant in the context of both the identification of transmembrane signaling partners for GPI-anchored T-cad as well as the demonstration of a novel mechanism whereby Grp78 can influence endothelial cell survival as a cell surface signaling receptor rather than an intracellular chaperone.

    Original languageEnglish
    Pages (from-to)4004-4017
    Number of pages14
    JournalMolecular and Cellular Biology
    Volume28
    Issue number12
    DOIs
    Publication statusPublished - 06-2008

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    Glycosylphosphatidylinositols
    Cell Survival
    Endothelial Cells
    Integrins
    Proteins
    Biotinylation
    Blocking Antibodies
    Cell Surface Receptors
    GABA-A Receptors
    Immunoprecipitation
    Confocal Microscopy
    Small Interfering RNA
    Signal Transduction
    Membrane Proteins
    High Pressure Liquid Chromatography
    Cell Membrane
    Lipids
    Glucose
    H-cadherin

    All Science Journal Classification (ASJC) codes

    • Molecular Biology
    • Genetics
    • Cell Biology

    Cite this

    Philippova, Maria ; Ivanov, Danila ; Joshi, Manjunath B. ; Kyriakakis, Emmanouil ; Rupp, Katharina ; Afonyushkin, Taras ; Bochkov, Valery ; Erne, Paul ; Resink, Therese J. / Identification of proteins associating with glycosylphosphatidylinositol- anchored T-cadherin on the surface of vascular endothelial cells : Role for Grp78/BiP in T-cadherin-dependent cell survival. In: Molecular and Cellular Biology. 2008 ; Vol. 28, No. 12. pp. 4004-4017.
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    abstract = "There is scant knowledge regarding how cell surface lipid-anchored T-cadherin (T-cad) transmits signals through the plasma membrane to its intracellular targets. This study aimed to identify membrane proteins colocalizing with atypical glycosylphosphatidylinositol (GPI)-anchored T-cad on the surface of endothelial cells and to evaluate their role as signaling adaptors for T-cad. Application of coimmunoprecipitation from endothelial cells expressing c-myc-tagged T-cad and high-performance liquid chromatography revealed putative association of T-cad with the following proteins: glucose-related protein GRP78, GABA-A receptor α1 subunit, integrin β3, and two hypothetical proteins, LOC124245 and FLJ32070. Association of Grp78 and integrin β3 with T-cad on the cell surface was confirmed by surface biotinylation and reciprocal immunoprecipitation and by confocal microscopy. Use of anti-Grp78 blocking antibodies, Grp78 small interfering RNA, and coexpression of constitutively active Akt demonstrated an essential role for surface Grp78 in T-cad-dependent survival signal transduction via Akt in endothelial cells. The findings herein are relevant in the context of both the identification of transmembrane signaling partners for GPI-anchored T-cad as well as the demonstration of a novel mechanism whereby Grp78 can influence endothelial cell survival as a cell surface signaling receptor rather than an intracellular chaperone.",
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    Identification of proteins associating with glycosylphosphatidylinositol- anchored T-cadherin on the surface of vascular endothelial cells : Role for Grp78/BiP in T-cadherin-dependent cell survival. / Philippova, Maria; Ivanov, Danila; Joshi, Manjunath B.; Kyriakakis, Emmanouil; Rupp, Katharina; Afonyushkin, Taras; Bochkov, Valery; Erne, Paul; Resink, Therese J.

    In: Molecular and Cellular Biology, Vol. 28, No. 12, 06.2008, p. 4004-4017.

    Research output: Contribution to journalArticle

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    AU - Philippova, Maria

    AU - Ivanov, Danila

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    AU - Resink, Therese J.

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