Probing subcellular iron availability with genetically encoded nitric oxide biosensors

Sevimli, Gülşah and Alston, Amy E. and Funk, Felix and Flühmann, Beat and Malli, Roland and Graier, Wolfgang F. and Eroğlu, Emrah (2022) Probing subcellular iron availability with genetically encoded nitric oxide biosensors. Biosensors, 12 (10). ISSN 2079-6374

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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.
Item Type: Article
Uncontrolled Keywords: carboxymaltose; cellular iron uptake; Ferinject; fluorescent biosensor; geNOps; HBED; iron sucrose; labile iron; Venofer
Divisions: Faculty of Engineering and Natural Sciences > Academic programs > Biological Sciences & Bio Eng.
Faculty of Engineering and Natural Sciences
Depositing User: Emrah Eroğlu
Date Deposited: 27 Mar 2023 12:18
Last Modified: 27 Mar 2023 12:18

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