Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz

Logo MUG-Forschungsportal

Gewählte Publikation:

SHR Neuro Krebs Kardio Lipid Stoffw Microb

Akyol, A; Çimen, Ş; Gottschalk, B; Erdoğan, YC; Lischnig, A; Barton, Alston, A; Digigow, R; Flühmann, B; Eroğlu, E; Graier, WF; Malli, R.
Tracking Intracellular Labile Iron with a Genetically Encoded Fluorescent Reporter System Based on Protein Stability.
ACS Sens. 2025; Doi: 10.1021/acssensors.5c01165 [OPEN ACCESS]
Web of Science PubMed PUBMED Central FullText FullText_MUG

 

Führende Autor*innen der Med Uni Graz
Akyol Ali
Malli Roland
Co-Autor*innen der Med Uni Graz
EROGLU Emrah
Gottschalk Benjamin
Graier Wolfgang
Altmetrics:

Dimensions Citations:

Plum Analytics:

Scite (citation analytics):

Abstract:
Here, we developed IronFist, a genetically encoded fluorescent reporter that enables dynamic tracking of labile ferrous ions (Fe2+) in live cells. IronFist is a bicistronic system combining the iron-sensitive hemerythrin-like (Hr) domain from the F-box and leucine-rich repeat protein 5 (FBXL5), fused to the bright fluorescent protein (FP) mNeonGreen, alongside mCherry as a reference FP signal. When labile iron levels are low, Hr-mNeonGreen undergoes ubiquitination and degradation, leading to a low green-to-red fluorescence ratio. Conversely, as cytosolic Fe2+ levels rise and Fe2+ ions bind to Hr, the green fluorescence is stabilized, increasing the IronFist ratio signal. Using IronFist for end point measurements, we observed that most cells maintain low basal labile iron levels. However, upon treatment with iron(II) sulfate or iron carbohydrate nanoparticles, we detected significant elevations in the cellular labile iron pool (LIP). Cells responded faster and more strongly to iron(II) sulfate, whereas responses to iron carbohydrate nanoparticles were slower and weaker. Time-lapse imaging further revealed substantial cell-to-cell heterogeneity in iron handling. We conclude that IronFist fills a critical methodological gap in assessing cellular iron homeostasis and related pathologies by enabling high-content tracking of iron dynamics at the single-cell level.

Find related publications in this database (Keywords)
iron reporter
iron carbohydrate nanoparticles
fluorescence microscopy
labile iron pool
Hr domain
iron-dependent degron
© Med Uni Graz • Impressum