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

Metallomics. 2017 May 24;9(5):525-534. doi: 10.1039/c6mt00291a.

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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Gel
  • Female
  • Fish Proteins / analysis*
  • Fish Proteins / metabolism
  • Metalloproteins / analysis*
  • Metalloproteins / metabolism
  • Models, Molecular
  • Ovary / drug effects
  • Ovary / metabolism*
  • Proteomics / methods
  • Spectrometry, Mass, Electrospray Ionization
  • Uranium / analysis*
  • Uranium / metabolism
  • Uranium / toxicity
  • Water Pollutants, Chemical / analysis*
  • Water Pollutants, Chemical / metabolism
  • Water Pollutants, Chemical / toxicity
  • Zebrafish

Substances

  • Fish Proteins
  • Metalloproteins
  • Water Pollutants, Chemical
  • Uranium