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In vivo identification of potential uranium protein targets in zebrafish ovaries after chronic waterborne exposure

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Abstract

Ecotoxicological studies have indicated the reprotoxicity of uranium (U) in zebrafish, but its molecular mechanisms remain unclear. Due to the non-covalent nature of U–protein complexes, canonical proteomics approaches are often not relevant as they usually use denaturating reagents or solvents. In this study, non-denaturating (ND) methods were used to obtain insight into the nature of U potential targets in ovaries of reproduced and non-reproduced zebrafish after 20 days of exposure to an environmentally relevant U concentration (20 μg L−1). After the ND sample preparation, 1-dimensional (SEC) and 2-dimensional (OGE × SEC) separations followed by ICP-sector-field MS measurements (U, P, Fe, Cu, and Zn) enabled the determination of chemical characteristics (MW, pI) of the metal–protein complexes. Phosphorus and U coelution confirmed the affinity of U for P-containing proteins. In addition, 2D separation allowed the discrimination of Fe-metalloproteins as potential U targets. Finally, 20 protein candidates for U complexation were identified after tryptic digestion conditions by LC-ESI FT MS and a database search. Potential U targets were mainly involved in three biological processes: oxidative stress regulation (SOD, GST), cytoskeleton structure (actin) and embryo early development (vtg, initiation factor).

Graphical abstract: In vivo identification of potential uranium protein targets in zebrafish ovaries after chronic waterborne exposure

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Publication details

The article was received on 09 Dec 2016, accepted on 22 Feb 2017 and first published on 20 Mar 2017


Article type: Paper
DOI: 10.1039/C6MT00291A
Citation: Metallomics, 2017, Advance Article
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    In vivo identification of potential uranium protein targets in zebrafish ovaries after chronic waterborne exposure

    Y. Eb-Levadoux, S. Frelon, O. Simon, C. Arnaudguilhem, R. Lobinski and S. Mounicou, Metallomics, 2017, Advance Article , DOI: 10.1039/C6MT00291A

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