Doppel and PrPC co-immunoprecipitate in detergent-resistant membrane domains of epithelial FRT cells. - Institut Pasteur Access content directly
Journal Articles Biochemical Journal Year : 2010

Doppel and PrPC co-immunoprecipitate in detergent-resistant membrane domains of epithelial FRT cells.

Abstract

Dpl (doppel) is a paralogue of the PrPC (cellular prion protein), whose misfolded conformer (the scrapie prion protein, PrPSc) is responsible for the onset of TSEs (transmissible spongiform encephalopathies) or prion diseases. It has been shown that the ectopic expression of Dpl in the brains of some lines of PrP-knockout mice provokes cerebellar ataxia, which can be rescued by the reintroduction of the PrP gene, suggesting a functional interaction between the two proteins. It is, however, still unclear where, and under which conditions, this event may occur. In the present study we addressed this issue by analysing the intracellular localization and the interaction between Dpl and PrPC in FRT (Fischer rat thyroid) cells stably expressing the two proteins separately or together. We show that both proteins localize prevalently on the basolateral surface of FRT cells, in both singly and doubly transfected clones. Interestingly we found that they associate with DRMs (detergent-resistant membranes) or lipid rafts, from where they can be co-immunoprecipitated in a cholesterol-dependent fashion. Although the interaction between Dpl and PrPC has been suggested before, our results provide the first clear evidence that this interaction occurs in rafts and is dependent on the integrity of these membrane microdomains. Furthermore, both Dpl and PrPC could be immunoprecipitated with flotillin-2, a raft protein involved in endocytosis and cell signalling events, suggesting that they share the same lipid environment.
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Dates and versions

pasteur-00490963 , version 1 (10-06-2010)

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Attribution - NonCommercial

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Anna Caputo, Daniela Sarnataro, Vincenza Campana, Maddalena Costanzo, Alessandro Negro, et al.. Doppel and PrPC co-immunoprecipitate in detergent-resistant membrane domains of epithelial FRT cells.. Biochemical Journal, 2010, 425 (2), pp.341-51. ⟨10.1042/BJ20091050⟩. ⟨pasteur-00490963⟩
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