title: Development of non-denaturing off-gel isoelectric focusing for the separation of uranium-protein complexes in fish.
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.
authors.affiliation: LCABIE - UMR5254, Technopôle Hélioparc Pau Pyrénées, 2 avenue du Président Angot, 64053, Pau Cedex 09, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Bucher
authors.forename: Sandrine
authors.initials: S
authors.lastname: Frelon
authors.forename: Olivier
authors.initials: O
authors.lastname: Simon
authors.forename: Ryszard
authors.initials: R
authors.lastname: Lobinski
authors.forename: Sandra
authors.initials: S
authors.lastname: Mounicou
mesh_terms: D000818:Animals; D002417:Cattle; D002623:Chemistry Techniques, Analytical; D003600:Cytosol / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007834:Lasers; D008670:Metals / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics; D018892:Proton-Motive Force; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013696:Temperature; D014501:Uranium / Q000737:chemistry*; D015027:Zebrafish
chemical_list: D008670:Metals; D011506:Proteins; D012710:Serum Albumin, Bovine; D014501:Uranium