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The Fe content of Jurkat cells grown on transferrin-bound iron (TBI) and Fe(III) citrate (FC) was characterized using Mössbauer, electron paramagnetic resonance, and UV-vis spectroscopies, as well as electron and inductively coupled plasma mass spectrometry. Isolated mitochondria were similarly characterized. Fe-limited cells contained ~100 μM essential Fe, mainly as mitochondrial Fe and nonmitochondrial non-heme high-spin Fe(II). Cells replete with Fe also contained ferritin-bound Fe and Fe(III) oxyhydroxide nanoparticles. Only 400 ± 100 Fe ions were loaded per ferritin complex, regardless of the growth medium Fe concentration. Ferritin regulation thus appears to be more complex than is commonly assumed. The magnetic and structural properties of Jurkat nanoparticles differed from those of yeast mitochondria. They were smaller and may be located in the cytosol. The extent of nanoparticle formation scaled nonlinearly with the concentration of Fe in the medium. Nanoparticle formation was not strongly correlated with reactive oxygen species (ROS) damage. Cells could utilize nanoparticle Fe, converting such aggregates into essential Fe forms. Cells grown on galactose rather than glucose respired faster, grew slower, exhibited more ROS damage, and generally contained more nanoparticles. Cells grown with TBI rather than FC contained less Fe overall, more ferritin, and fewer nanoparticles. Cells in which the level of transferrin receptor expression was increased contained more ferritin Fe. Frataxin-deficient cells contained more nanoparticles than comparable wild-type cells. Data were analyzed by a chemically based mathematical model. Although simple, it captured essential features of Fe import, trafficking, and regulation. TBI import was highly regulated, but FC import was not. Nanoparticle formation was not regulated, but the rate was third-order in cytosolic Fe.

Mössbauer Study and Modeling of Iron Import and Trafficking in Human Jurkat Cells