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Development of non-denaturing off-gel isoelectric focusing for the separation of uranium–protein complexes in fish

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Abstract

An off-gel non-denaturing isoelectric focusing (IEF) method was developed to separate uranium–biomolecule complexes from biological samples as a first step in a multidimensional metalloproteomic approach. Analysis of a synthetic uranium–bovine serum albumin complex demonstrated the focusing ability of the liquid-phase IEF method and the preservation of most of the uranium–protein interactions. The developed method was applied to gill cytosol prepared from zebrafish (Danio rerio) exposed to depleted uranium. The results were compared in terms of resolution, recovery, and protein identities with those obtained by in-gel IEF using an immobilized pH gradient gel strip.

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Acknowledgments

The authors would like to thank the French Institute for Radiological Protection and Nuclear Safety (IRSN), Conseil Régional Aquitain, and the French National Research Agency (ANR) with the ST MALO - 2010 JCJC 713 1 project for the financial support of this project.

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Authors and Affiliations

  1. LCABIE - UMR5254, Technopôle Hélioparc Pau Pyrénées, 2 avenue du Président Angot, 64053, Pau Cedex 09, France

    Guillaume Bucher, Ryszard Lobinski & Sandra Mounicou

  2. IRSN/PRP-ENV/SERIS - Laboratoire de Biogéochimie, Biodisponibilité et Transferts des radionucléides - BP3, 13115, St Paul lez Durance Cedex, France

    Guillaume Bucher, Sandrine Frelon & Olivier Simon

Authors
  1. Guillaume Bucher
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  2. Sandrine Frelon
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  3. Olivier Simon
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  4. Ryszard Lobinski
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  5. Sandra Mounicou
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Corresponding author

Correspondence to Sandra Mounicou.

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Bucher, G., Frelon, S., Simon, O. et al. Development of non-denaturing off-gel isoelectric focusing for the separation of uranium–protein complexes in fish. Anal Bioanal Chem 406, 3517–3520 (2014). https://doi.org/10.1007/s00216-014-7768-x

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  • DOI: https://doi.org/10.1007/s00216-014-7768-x

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