Deciphering the uranium target proteins in human dopaminergic SH-SY5Y cells

Claude Vidaud 1 Mélanie Robert 1 Eduardo Paredes 2 Richard Ortega 3 Emilie Avazeri 1 Lun Jing 4 Jean-Marie Guigonis 4 Carole Bresson 2 Veronique Malard 1, 5, *
* Corresponding author
4 TIRO - UMR E4320 - Transporteurs en Imagerie et Radiothérapie en Oncologie
CEA - Commissariat à l'énergie atomique et aux énergies alternatives : DRF/BIAM, CNRS - Centre National de la Recherche Scientifique, TIRO-MATOs - UMR E4320 : UMR E4320
5 IPM - Interactions Protéine Métal
BIAM - Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) : DRF/BIAM
Abstract : Uranium (U) is the heaviest naturally occurring element ubiquitously present in the Earth’s crust. Human exposure to low levels of U is, therefore, unavoidable. Recently, several studies have clearly pointed out that the brain is a sensitive target for U, but the mechanisms leading to the observed neurological alterations are not fully known. To deepen our knowledge of the biochemical disturbances resulting from U(VI) toxicity in neuronal cells, two complementary strategies were set up to identify the proteins that selectively bind U(VI) in human dopaminergic SH-SY5Y cells. The first strategy relies on the selective capture of proteins capable of binding U(VI), using immobilized metal affinity chromatography, and starting from lysates of cells grown in a U(VI)-free medium. The second strategy is based on the separation of U-enriched protein fractions by size-exclusion chromatography, starting from lysates of U(VI)-exposed cells. High-resolution mass spectrometry helped us to highlight 269 common proteins identified as the urano-proteome. They were further analyzed to characterize their cellular localization and biological functions. Four canonical pathways, related to the protein ubiquitination system, gluconeogenesis, glycolysis, and the actin cytoskeleton proteins, were particularly emphasized due to their high content of U(VI)-bound proteins. A semi-quantification was performed to concentrate on the ten most abundant proteins, whose physico-chemical characteristics were studied in particular depth. The selective interaction of U(VI) with these proteins is an initial element of proof of the possible metabolic effects of U(VI) on neuronal cells at the molecular level.
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Journal articles
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Submitted on : Tuesday, June 25, 2019 - 11:21:56 AM
Last modification on : Thursday, June 27, 2019 - 1:25:24 AM

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Claude Vidaud, Mélanie Robert, Eduardo Paredes, Richard Ortega, Emilie Avazeri, et al.. Deciphering the uranium target proteins in human dopaminergic SH-SY5Y cells. Archives of Toxicology, Springer Verlag, 2019, ⟨10.1007/s00204-019-02497-4⟩. ⟨cea-02164621⟩

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