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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Acknowledgements
We acknowledge the Transversal Toxicology Program run by the Direction de la Recherche Fondamentale (DRF) of the Commissariat à l’énergie atomique et aux énergies alternatives (CEA) and the financial support from this program. E.P. also thanks the CEA Enhanced Eurotalents program, co-funded by the European Commission through the Marie Sklodowska-Curie COFUND program under the 7th Framework Program for Research and Technological Development. This work was also partially funded by the CNRS Interdisciplinary Mission through the PEPS (Projet Exploratoire Premier Soutien) Faidora program (Faibles Doses, Risques, Alertes). We acknowledge the Li2D laboratory for its technical support.
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Vidaud, C., Robert, M., Paredes, E. et al. Deciphering the uranium target proteins in human dopaminergic SH-SY5Y cells. Arch Toxicol 93, 2141–2154 (2019). https://doi.org/10.1007/s00204-019-02497-4
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DOI: https://doi.org/10.1007/s00204-019-02497-4