title: Specific capture of uranyl protein targets by metal affinity chromatography.
abstract: To improve general understanding of biochemical mechanisms in the field of uranium toxicology, the identification of protein targets needs to be intensified. Immobilized metal affinity chromatography (IMAC) has been widely developed as a powerful tool for capturing metal binding proteins from biological extracts. However uranyl cations (UO2(2+)) have particular physico-chemical characteristics which prevent them from being immobilized on classical metal chelating supports. We report here on the first development of an immobilized uranyl affinity chromatography method, based on the cation-exchange properties of aminophosphonate groups for uranyl binding. The cation distribution coefficient and loading capacity on the support were determined. Then the stability of the uranyl-bonded phase under our chromatographic conditions was optimized to promote affinity mechanisms. The successful enrichment of uranyl binding proteins from human serum was then proven using proteomic and mass spectral analysis.
authors.affiliation: CEA Valrhô, DSV/IBEB/Service de Biochimie et de Toxicologie Nucléaire, B.P. 17171, F-30207 Bagnols sur Cèze, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Basset
authors.forename: Alain
authors.initials: A
authors.lastname: Dedieu
authors.forename: Philippe
authors.initials: P
authors.lastname: Guérin
authors.forename: Eric
authors.initials: E
authors.lastname: Quéméneur
authors.forename: Daniel
authors.initials: D
authors.lastname: Meyer
authors.forename: Claude
authors.initials: C
authors.lastname: Vidaud
mesh_terms: D001798:Blood Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D002411:Cation Exchange Resins; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D009942:Organometallic Compounds / Q000378:metabolism*; D011485:Protein Binding; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014501:Uranium / Q000378:metabolism*
chemical_list: D001798:Blood Proteins; D002411:Cation Exchange Resins; D009942:Organometallic Compounds; C005460:uranyl acetate; D014501:Uranium