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dc.contributor.authorArulpragasam, Ajanthy
dc.contributor.authorMagno, A.
dc.contributor.authorIngley, E.
dc.contributor.authorBrown, S.
dc.contributor.authorConigrave, A.
dc.contributor.authorRatajczak, T.
dc.contributor.authorWard, B.
dc.date.accessioned2017-01-30T13:22:58Z
dc.date.available2017-01-30T13:22:58Z
dc.date.created2015-10-29T04:10:02Z
dc.date.issued2012
dc.identifier.citationArulpragasam, 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.
dc.identifier.urihttp://hdl.handle.net/20.500.11937/31034
dc.identifier.doi10.1042/BJ20111277
dc.description.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.

dc.titleThe adaptor protein 14-3-3 binds to the calcium-sensing receptor and attenuates receptor-mediated Rho kinase signalling
dc.typeJournal Article
dcterms.source.volume441
dcterms.source.number3
dcterms.source.startPage995
dcterms.source.endPage1006
dcterms.source.issn0264-6021
dcterms.source.titleBiochemical Journal
curtin.departmentSchool of Biomedical Sciences
curtin.accessStatusOpen access via publisher


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