Metabolism of iron in amoebas

Ferritins are complex protein structures for iron storage and detoxification. They are typically found in the cytoplasm of the cell; however, they were also identified in mitochondria and chloroplasts. They are distributed among all living organisms but uncommon in non-photosynthetic protists. Surprisingly, a ferritin gene was found in the genome of free-living amoeba Naegleria gruberi. Bioinformatic analysis confirms that it belongs to a group of eukaryotic ferritins. Upon creating antibodies in a laboratory rat, we localized the protein in mitochondria of N. gruberi. The native recombinant protein without the mitochondrial presequence was used for further in vitro experiments. Using growth experiments, we established that the expression of ferritin is dependent on a concentration of iron in cultivation media; however, we were not able to directly prove its ability to bind iron in vitro. We concluded that the function of the ferritin of N. gruberi is most likely related to metabolism of iron. Side experiments were focused on a different protein from the ferritin family - Dps protein in Acanthamoeba castellanii. Despite of a successful creation of antibodies, we were not able to detect the expression of protein neither in total cell lysates nor in subcellular fractions of acanthamoebas under..