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A reactivity-based probe of the intracellular labile ferrous iron pool.

Nat Chem Biol. 2016 Sep;12(9):680-5

Authors: Spangler B, Morgan CW, Fontaine SD, Vander Wal MN, Chang CJ, Wells JA, Renslo AR

Improved methods for studying intracellular reactive Fe(II) are of significant interest for studies of iron metabolism and disease-relevant changes in iron homeostasis. Here we describe a highly selective reactivity-based probe in which a Fenton-type reaction with intracellular labile Fe(II) leads to unmasking of the aminonucleoside puromycin. Puromycin leaves a permanent and dose-dependent mark on treated cells that can be detected with high sensitivity and precision using a high-content, plate-based immunofluorescence assay. Using this new probe and screening approach, we detected alteration of cellular labile Fe(II) in response extracellular iron conditioning, overexpression of iron storage and/or export proteins, and post-translational regulation of iron export. We also used this new tool to demonstrate that labile Fe(II) pools are larger in cancer cells than in nontumorigenic cells.

PMID: 27376690 [PubMed – indexed for MEDLINE]