Uranium (U) is a naturally occurring radionuclide. The redistribution of U in the environment is primarily driven by anthropogenic activities, including mining operations, the nuclear industry, military activities, and soil fertilization. Local accumulation of U can pose potential risks to ecosystems, agricultural systems, and ultimately human health. Although U is not essential for plants, yet they take it up from the soil and incorporate it into their biomass. This eventually enters the food chain, posing a significant health risk to humans. To identify proteins that could be cellular targets for U in planta, we used a metalloproteomic approach combining column chromatographic fractionation analyses with protein identification by high-resolution mass spectrometry (MS)-based shotgun proteomics.