title: Characterization of UO2(2+) binding to osteopontin, a highly phosphorylated protein: insights into potential mechanisms of uranyl accumulation in bones.
abstract: Bones are one of the few organs in which uranyl (UO2(2+)) accumulates. This large dioxo-cation displays affinity for carboxylates, phenolates and phosphorylated functional groups in proteins. The noncollagenous protein osteopontin (OPN) plays an important role in bone homeostasis. It is mainly found in the extracellular matrix of mineralized tissues but also in body fluids such as milk, blood and urine. Furthermore, OPN is an intrinsically disordered protein, which, like other proteins of the SIBLING family, contains a polyaspartic acid sequence and numerous patterns of alternating acidic and phosphorylated residues. All these properties led to the hypothesis that this protein could be prone to UO2(2+) binding. In this work, a simple purification procedure enabling highly purified bovine (bOPN) and human OPN (hOPN) to be obtained was developed. Various biophysical approaches were set up to study the impact of phosphorylations on the affinity of OPN for UO2(2+) as well as the formation of stable complexes originating from structural changes induced by the binding of this metal cation. The results obtained suggest a new mechanism of the interaction of UO2(2+) with bone metabolism and a new role for OPN as a metal transporter.
authors: CEA/DSV/iBEB/SBTN, Laboratoire d'Etude des Protéines Cibles, BP 17171, 30207 Bagnols sur Cèze Cédex, France. claude.vidaud@cea.fr.
authors: Lei
authors: L
authors: Qi
authors: Christian
authors: C
authors: Basset
authors: Olivier
authors: O
authors: Averseng
authors: Eric
authors: E
authors: Quéméneur
authors: Agnès
authors: A
authors: Hagège
authors: Claude
authors: C
authors: Vidaud
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D001842:Bone and Bones / Q000378:metabolism*; D002417:Cattle; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D053495:Osteopontin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D014501:Uranium / Q000378:metabolism*; D017974:Uranium Compounds / Q000378:metabolism*
chemical_list: D017974:Uranium Compounds; D053495:Osteopontin; D014501:Uranium; C012597:uranium dioxide