Image-based quantification of mitochondrial iron uptake via Mitoferrin-2
Creators
- 1. CSL Vifor, St. Gallen, Switzerland
- 2. CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria
- 3. Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria
Description
Iron is a trace element that is critical for most living organisms and plays a key role in a wide variety of metabolic processes. In the mitochondrion, iron is involved in producing iron-sulfur clusters and synthesis of heme and kept within physiological ranges by concerted activity of multiple molecules. Mitochondrial iron uptake is mediated by the solute carrier transporters Mitoferrin-1 (SLC25A37) and Mitoferrin-2 (SLC25A28). While Mitoferrin-1 is mainly involved in erythropoiesis, the cellular function of the ubiquitously expressed Mitoferrin-2 remains less well defined. Furthermore, Mitoferrin-2 is associated with several human diseases, including cancer, cardiovascular and metabolic diseases, hence representing a potential therapeutic target. Here, we developed a robust approach to quantify mitochondrial iron uptake mediated by Mitoferrin-2 in living cells. We utilize HEK293 cells with inducible expression of Mitoferrin-2 and measure iron-induced quenching of rhodamine B[(1,10-
phenanthroline-5-yl)-aminocarbonyl]benzyl ester (RPA) fluorescence and validate this assay for mediumthroughput screening. This assay may allow identification and characterization of Mitoferrin-2 modulators and could enable drug discovery for this target.
Files
Polesel_2024.pdf
Files
(1.4 MB)
| Name | Size | Download all |
|---|---|---|
|
md5:583875ea9d959da0a26bc2fd888bbbdf
|
1.4 MB | Preview Download |
Additional details
Identifiers
Related works
- Is identical to
- 10.1016/j.mito.2024.101889 (DOI)