title: Impact of uranium (U) on the cellular glutathione pool and resultant consequences for the redox status of U.
abstract: Uranium (U) as a redox-active heavy metal can cause various redox imbalances in plant cells. Measurements of the cellular glutathione/glutathione disulfide (GSH/GSSG) by HPLC after cellular U contact revealed an interference with this essential redox couple. The GSH content remained unaffected by 10 μM U whereas the GSSG level immediately increased. In contrast, higher U concentrations (50 μM) drastically raised both forms. Using the Nernst equation, it was possible to calculate the half-cell reduction potential of 2GSH/GSSG. In case of lower U contents the cellular redox environment shifted towards more oxidizing conditions whereas the opposite effect was obtained by higher U contents. This indicates that U contact causes a consumption of reduced redox equivalents. Artificial depletion of GSH by chlorodinitrobenzene and measuring the cellular reducing capacity by tetrazolium salt reduction underlined the strong requirement of reduced redox equivalents. An additional element of cellular U detoxification mechanisms is the complex formation between the heavy metal and carboxylic functionalities of GSH. Because two GSH molecules catalyze electron transfers each with one electron forming a dimer (GSSG) two UO(2)                                   (2+) are reduced to each UO(2)                                   (+) by unbound redox sensitive sulfhydryl moieties. UO(2)                                   (+) subsequently disproportionates to UO(2)                                   (2+) and U(4+). This explains that in vitro experiments revealed a reduction to U(IV) of only around 33% of initial U(VI). Cellular U(IV) was transiently detected with the highest level after 2 h of U contact. Hence, it can be proposed that these reducing processes are an important element of defense reactions induced by this heavy metal.
authors.affiliation: Biogeochemistry Division, Institute of Radiochemistry, Helmholtz-Zentrum Dresden-Rossendorf eV, Dresden, Germany. k.viehweger@hzdr.de
authors.forename: Katrin
authors.initials: K
authors.lastname: Viehweger
authors.forename: Gerhard
authors.initials: G
authors.lastname: Geipel
authors.forename: Gert
authors.initials: G
authors.lastname: Bernhard
mesh_terms: D029688:Brassica napus / Q000166:cytology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D004137:Dinitrochlorobenzene / Q000737:chemistry; D005978:Glutathione / Q000378:metabolism*; D010084:Oxidation-Reduction; D014501:Uranium / Q000378:metabolism*
chemical_list: D004137:Dinitrochlorobenzene; D014501:Uranium; D005978:Glutathione