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    The adaptor protein 14-3-3 binds to the calcium-sensing receptor and attenuates receptor-mediated Rho kinase signalling

    Access Status
    Open access via publisher
    Authors
    Arulpragasam, Ajanthy
    Magno, A.
    Ingley, E.
    Brown, S.
    Conigrave, A.
    Ratajczak, T.
    Ward, B.
    Date
    2012
    Type
    Journal Article
    
    Metadata
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    Citation
    Arulpragasam, A. and Magno, A. and Ingley, E. and Brown, S. and Conigrave, A. and Ratajczak, T. and Ward, B. 2012. The adaptor protein 14-3-3 binds to the calcium-sensing receptor and attenuates receptor-mediated Rho kinase signalling. Biochemical Journal. 441 (3): pp. 995-1006.
    Source Title
    Biochemical Journal
    DOI
    10.1042/BJ20111277
    ISSN
    0264-6021
    School
    School of Biomedical Sciences
    URI
    http://hdl.handle.net/20.500.11937/31034
    Collection
    • Curtin Research Publications
    Abstract

    A yeast two-hybrid screen performed to identify binding partners of the CaR (calcium-sensing receptor) intracellular tail identified the adaptor protein 14-3-3? as a novel binding partner that bound to the proximal membrane region important for CaR expression and signalling. The 14-3-3? protein directly interacted with the CaR tail in pull-down studies and FLAG-tagged CaR co-immunoprecipitated with EGFP (enhanced green fluorescent protein)-tagged 14-3-3? when co-expressed in HEK (human embryonic kidney)-293 or COS-1 cells. The interaction between the CaR and 14-3-3? did not require a putative binding site in the membrane-proximal region of the CaR tail and was independent of PKC (protein kinase C) phosphorylation. Confocal microscopy demonstrated co-localization of the CaR and EGFP-14-3-3? in the ER (endoplasmic reticulum) of HEK-293 cells that stably expressed the CaR (HEK-293/CaR cells), but 14-3-3? overexpression had no effect on membrane expression of the CaR. Overexpression of 14-3-3? in HEK-293/CaR cells attenuated CaR-mediated Rho signalling, but had no effect on ERK (extracellular-signal-regulated kinase) 1/2 signalling. Another isoform identified from the library, 14-3-3?, exhibited similar behaviour to that of 14-3-3? with respect to CaR tail binding, cellular co-localization and impact on receptor-mediated signalling. However, unlike 14-3-3?, this isoform, when overexpressed, significantly reduced CaR plasma membrane expression. Results indicate that 14-3-3 proteins mediate CaR-dependent Rho signalling and may modulate the plasma membrane expression of the CaR. © The Authors Journal compilation © 2012 Biochemical Society.

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