title: Ascorbate-induced high-affinity binding of copper to cytosolic proteins.
abstract: Copper chaperones are necessary for intracellular trafficking of copper to target proteins. This is probably because the milieu inside the cell has a large capacity for sequestering this metal. By fluorometry using a fluorescent Cu(II) chelator and by centrifugal ultrafiltration, we have studied copper binding of the whole cytosolic proteins from mouse brain and liver, and found that their binding capacity and affinity for copper were markedly increased by ascorbate. Brain cytosolic protein bound, with high affinity, 63 nmol of copper/mg, more than half of which was redox-inactive, as indicated by its inability to catalyze oxidation of ascorbate. Most of the bound copper was in the Cu(I) state, coordinating to thiol groups of protein. Cytosolic protein competed for copper more strongly than GSH when compared at their relative concentrations in tissues. The results taken together suggest that protein thiols of cytosol can strongly sequester copper.
authors.affiliation: Department of Biochemistry, Wakayama Medical University, 811-1 Kimiidera, Wakayama 641-0012, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Ohta
authors.forename: N
authors.initials: N
authors.lastname: Shiraishi
authors.forename: Y
authors.initials: Y
authors.lastname: Inai
authors.forename: I S
authors.initials: IS
authors.lastname: Lee
authors.forename: H
authors.initials: H
authors.lastname: Iwahashi
authors.forename: M
authors.initials: M
authors.lastname: Nishikimi
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology*; D001798:Blood Proteins / Q000378:metabolism; D001921:Brain / Q000187:drug effects*; D001923:Brain Chemistry; D002614:Chelating Agents / Q000378:metabolism; D003300:Copper / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000187:drug effects*; D051379:Mice; D010084:Oxidation-Reduction; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D000975:Antioxidants; D001798:Blood Proteins; D002614:Chelating Agents; D005452:Fluoresceins; D005456:Fluorescent Dyes; D011506:Proteins; D003300:Copper; D001205:Ascorbic Acid