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AbstractThe biosynthesis of selenoproteins was studied in relation to milk formation and mammary cell biology by incubating the bovine mammary cell line MAC-T with ((75)Se)selenite. Intracellular proteins and proteins secreted into the cell culture medium were separated by 2D electrophoresis, the selenoproteins were detected by autoradiography, and the proteins were identified by MALDI-TOF. Approximately 35 (75)Se-containing spots were found in the cell proteins from MAC-T cells. Among them, one-third showed high intensity. The strongest spot was identified as glutathione peroxidase 1. About 20 spots were observed in protein precipitated from cell culture medium, one-third of them being distinctly visible. In an attempt to study a perturbation of the system, the effect of retinoic acid (RA) on the formation of selenoproteins was investigated. The concentration of (75)Se in total cell protein was reduced by about 35% in cells cultured with RA compared with control cells, while the opposite effect was observed in protein precipitated from cell culture medium, which contained 60% more (75)Se in RA-treated samples than in controls. There were also indications that RA might affect different selenoproteins in different ways. The methods described provide a promising approach for further studies of the regulation of selenoprotein formation in the mammary gland.
CitationJ. Trace Elem. Med. Biol. 2008, 22 (3):224-233
JournalJournal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements (GMS)
SponsorsThis study was supported by grants from the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning, the Lund University Hospital, the Danish Dairy Research Foundation, the Påhlsson Foundation, and the SWEGENE Proteomics Centre. We would like to thank Liselotte Andersson, Ulrika Brynnel, Annette K. Nielsen, and Anna-Karin Påhlman for their skilled technical assistance, and Kristen Sejrsen for helpful advice. Biomedical Nutrition is a member of the EU NoE, The European Nutrigenomics Organisation (NuGO, no. 505944), and the NoE, Environmental Cancer risk, Nutrition and Individual Susceptibility (ECNIS, no. 513943), which partly supported the study.
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