Identification of potential uranium-binding proteins in Arabidopsis thaliana by a 2-dimensional gel electrophoresis-based approach

Uranium is metal that appears more and more as a soil contaminant, on the one hand because of the increased extraction of uranium worldwide for nuclear energy production, and on the other hand as a contaminant of phosphate fertilizers. Uranium is known to alter plant physiology, but the molecular actors implied in these alterations, i.e. the uranium-binding proteins, are poorly known up to now. In order to improve our knowledge on this topic, a 2D gel-based approach relying on protein correlation profiling was used. To this purpose, Arabidopsis thaliana cells were cultivated under photosynthetic conditions until a sufficient number of cells was reached. The cells were then rinsed and exposed to 50µM uranyl nitrate in a Murashige and Skoog medium deprived of phosphate or containing 30µM phosphate. The cells were then washed with 10mM sodium carbonate, twice with distilled water and kept frozen until use.