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

Abstract:

: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.

journal_name

Nat Chem Biol

journal_title

Nature chemical biology

authors

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

doi

10.1038/nchembio.2116

subject

Has Abstract

pub_date

2016-09-01 00:00:00

pages

680-5

issue

9

eissn

1552-4450

issn

1552-4469

pii

nchembio.2116

journal_volume

12

pub_type

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