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

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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.

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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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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T1 - Identification of proteins associating with glycosylphosphatidylinositol- anchored T-cadherin on the surface of vascular endothelial cells

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AU - Philippova, Maria

AU - Ivanov, Danila

AU - Joshi, Manjunath B.

AU - Kyriakakis, Emmanouil

AU - Rupp, Katharina

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AU - Erne, Paul

AU - Resink, Therese J.

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