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

ABSTRACT: 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.

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ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Cell Culture

SUBMITTER: Hélène Diemer

LAB HEAD: Sarah CIANFERANI

PROVIDER: PXD056391 | Pride | 2026-02-09

REPOSITORIES: Pride

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Project description:Evolution of multicellular life forms has involved adaptation of organs that consist of multiple cell types, each with unique functional properties that as a collection, achieve complex organ function. Since each cell type is adapted to deliver specific functionality within the context of an organ, knowledge on functional landscapes occupied by individual cell types could improve comprehension of organ function at the molecular level. In kidney, podocytes and tubules are two cell types that work together, each with vastly different functional roles. Podocytes envelop the blood vessels in the glomerulus and act as filters while tubules, located downstream of the glomerulus, are responsible for reabsorption of important nutrients. Mitochondria hold a critical and well-studied role in tubules due to the high energetic requirements required to fulfill their function. In podocytes however, questions remain regarding the relevance of mitochondrial function in both normal physiology and pathology . Quantitative cross-linking mass spectrometry and proteomics together with a transgenic mitochondrial tagging strategy were used to investigate kidney cell-type specificity of mitochondria. These efforts revealed that despite similarities of podocyte and tubule mitochondrial proteomes, each contain unique features corresponding to known distinct functional roles. These include increased demand for energy production through the TCA cycle in tubules and increased detoxification demand in podocytes. Moreover, tubule and podocyte mitochondrial interactome differences revealed additional cell-type specific functional insights with alterations in betaine metabolism, lysine degradation, and other pathways not regulated through proteome abundance levels. Most importantly, these efforts illustrate that cell specific mitochondrial interactome differences within an organ can now be visualized. Therefore, this approach can generally be used to map cell-specific mitochondrial changes in disease, aging or even with therapy to better understand the roles and contributions of each cell type in normal physiology and pathology within an organ in ways not previously possible .

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

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