Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Sevimli, G; Alston, AE; Funk, F; Flühmann, B; Malli, R; Graier, WF; Eroglu, E.
Probing Subcellular Iron Availability with Genetically Encoded Nitric Oxide Biosensors.
Biosensors (Basel). 2022; 12(10):
Doi: 10.3390/bios12100903
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- Führende Autor*innen der Med Uni Graz
- EROGLU Emrah
- Graier Wolfgang
- Malli Roland
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- Abstract:
- Cellular iron supply is required for various biochemical processes. Measuring bioavailable iron in cells aids in obtaining a better understanding of its biochemical activities but is technically challenging. Existing techniques have several constraints that make precise localization difficult, and the lack of a functional readout makes it unclear whether the tested labile iron is available for metalloproteins. Here, we use geNOps; a ferrous iron-dependent genetically encoded fluorescent nitric oxide (NO) biosensor, to measure available iron in cellular locales. We exploited the nitrosylation-dependent fluorescence quenching of geNOps as a direct readout for cellular iron absorption, distribution, and availability. Our findings show that, in addition to ferrous iron salts, the complex of iron (III) with N,N'-bis (2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED) can activate the iron (II)-dependent NO probe within intact cells. Cell treatment for only 20 min with iron sucrose was also sufficient to activate the biosensor in the cytosol and mitochondria significantly; however, ferric carboxymaltose failed to functionalize the probe, even after 2 h of cell treatment. Our findings show that the geNOps approach detects available iron (II) in cultured cells and can be applied to assay functional iron (II) at the (sub)cellular level.
- Find related publications in this database (Keywords)
- labile iron
- cellular iron uptake
- fluorescent biosensor
- geNOps
- Ferinject
- Venofer
- carboxymaltose
- iron sucrose
- HBED