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Detection of intracellular iron by its regulatory effect

Item Type:Article
Title:Detection of intracellular iron by its regulatory effect
Creators Name:Li, J.Y. and Ram, G. and Gast, K. and Chen, X. and Barasch, K. and Mori, K. and Schmidt-Ott, K. and Wang, J. and Kuo, H.C. and Savage-Dunn, C. and Garrick, M.D. and Barasch, J.
Abstract: Intracellular iron regulates gene expression by inhibiting the interaction of iron regulatory proteins (IRPs) with RNA motifs called iron-responsive elements (IREs). To assay this interaction in living cells we have developed two fluorescent IRE-based reporters that rapidly, reversibly, and specifically respond to changes in cellular iron status as well as signaling that modifies IRP activity. The reporters were also sufficiently sensitive to distinguish apo- from holotransferrin in the medium, to detect the effect of modifiers of the transferrin pathway such as HFE, and to detect the donation or chelation of iron by siderophores bound to the lipocalin neutrophil gelatinase-associated lipocalin (Ngal). In addition, alternative configurations of the IRE motif either enhanced or repressed fluorescence, permitting a ratio analysis of the iron-dependent response. These characteristics make it possible to visualize iron-IRP-IRE interactions in vivo.
Keywords:Iron Regulatory Proteins, Iron-Responsive Element, Labile Iron Pool, Transferrin, HFE, Neutrophil Gelatinase-Associated Lipocalin, Siderophore
Source:American Journal of Physiology Cell Physiology
Publisher:American Physiological Society
Page Range:C1547-C1549
Date:December 2004
Official Publication:https://doi.org/10.1152/ajpcell.00260.2004
PubMed:View item in PubMed

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