Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levels
Iron is an essential metal for cellular metabolism and signaling, but it has adverse effects in excess. The physiological consequences of iron deficiency are well established, yet the relationship between iron supplementation and pericellular oxygen levels in cultured cells and their downstream effe...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-07-01
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| Series: | Redox Biology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S221323172200091X |
| _version_ | 1811257085248143360 |
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| author | Gulsah Sevimli Matthew J. Smith Tuba Akgul Caglar Şükriye Bilir Melike Secilmis Hamza Y. Altun Esra N. Yigit Fan Yang Thomas P. Keeley Roland Malli Gürkan Öztürk Giovanni E. Mann Emrah Eroglu |
| author_facet | Gulsah Sevimli Matthew J. Smith Tuba Akgul Caglar Şükriye Bilir Melike Secilmis Hamza Y. Altun Esra N. Yigit Fan Yang Thomas P. Keeley Roland Malli Gürkan Öztürk Giovanni E. Mann Emrah Eroglu |
| author_sort | Gulsah Sevimli |
| collection | DOAJ |
| description | Iron is an essential metal for cellular metabolism and signaling, but it has adverse effects in excess. The physiological consequences of iron deficiency are well established, yet the relationship between iron supplementation and pericellular oxygen levels in cultured cells and their downstream effects on metalloproteins has been less explored. This study exploits the metalloprotein geNOps in cultured HEK293T epithelial and EA.hy926 endothelial cells to test the iron-dependency in cells adapted to standard room air (18 kPa O2) or physiological normoxia (5 kPa O2). We show that cells in culture require iron supplementation to activate the metalloprotein geNOps and demonstrate for the first time that cells adapted to physiological normoxia require significantly lower iron compared to cells adapted to hyperoxia. This study establishes an essential role for recapitulating oxygen levels in vivo and uncovers a previously unrecognized requirement for ferrous iron supplementation under standard cell culture conditions to achieve geNOps functionality. |
| first_indexed | 2024-04-12T17:51:12Z |
| format | Article |
| id | doaj.art-ae43eb1dd385499690bf5af27e5b1a1b |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-04-12T17:51:12Z |
| publishDate | 2022-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj.art-ae43eb1dd385499690bf5af27e5b1a1b2022-12-22T03:22:30ZengElsevierRedox Biology2213-23172022-07-0153102319Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levelsGulsah Sevimli0Matthew J. Smith1Tuba Akgul Caglar2Şükriye Bilir3Melike Secilmis4Hamza Y. Altun5Esra N. Yigit6Fan Yang7Thomas P. Keeley8Roland Malli9Gürkan Öztürk10Giovanni E. Mann11Emrah Eroglu12Molecular Biology, Genetics and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, TurkeyKing's British Heart Foundation Centre of Research Excellence, School of Cardiovascular and Metabolic Medicine & Sciences, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London, SE1 9NH, UKMolecular Biology, Genetics and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, Turkey; Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, TurkeyResearch Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, TurkeyMolecular Biology, Genetics and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, TurkeyMolecular Biology, Genetics and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, TurkeyResearch Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey; Department of Biotechnology, Gebze Technical University, Kocaeli, TurkeyKing's British Heart Foundation Centre of Research Excellence, School of Cardiovascular and Metabolic Medicine & Sciences, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London, SE1 9NH, UKTarget Discovery Institute, University of Oxford, Oxford, OX3 7FZ, UKMolecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, 8010, Graz, Austria; BioTechMed Graz, Mozartgasse 12/II, 8010, Graz, AustriaResearch Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey; Physiology Department, International School of Medicine, Istanbul Medipol University, Istanbul, TurkeyKing's British Heart Foundation Centre of Research Excellence, School of Cardiovascular and Metabolic Medicine & Sciences, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London, SE1 9NH, UK; Corresponding author.Molecular Biology, Genetics and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, Turkey; Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey; Corresponding author. Molecular Biology, Genetics and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, Turkey.Iron is an essential metal for cellular metabolism and signaling, but it has adverse effects in excess. The physiological consequences of iron deficiency are well established, yet the relationship between iron supplementation and pericellular oxygen levels in cultured cells and their downstream effects on metalloproteins has been less explored. This study exploits the metalloprotein geNOps in cultured HEK293T epithelial and EA.hy926 endothelial cells to test the iron-dependency in cells adapted to standard room air (18 kPa O2) or physiological normoxia (5 kPa O2). We show that cells in culture require iron supplementation to activate the metalloprotein geNOps and demonstrate for the first time that cells adapted to physiological normoxia require significantly lower iron compared to cells adapted to hyperoxia. This study establishes an essential role for recapitulating oxygen levels in vivo and uncovers a previously unrecognized requirement for ferrous iron supplementation under standard cell culture conditions to achieve geNOps functionality.http://www.sciencedirect.com/science/article/pii/S221323172200091XPericellular oxygenHyperoxiaNormoxiaNO bioavailabilitygeNOpsFerrous iron |
| spellingShingle | Gulsah Sevimli Matthew J. Smith Tuba Akgul Caglar Şükriye Bilir Melike Secilmis Hamza Y. Altun Esra N. Yigit Fan Yang Thomas P. Keeley Roland Malli Gürkan Öztürk Giovanni E. Mann Emrah Eroglu Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levels Redox Biology Pericellular oxygen Hyperoxia Normoxia NO bioavailability geNOps Ferrous iron |
| title | Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levels |
| title_full | Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levels |
| title_fullStr | Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levels |
| title_full_unstemmed | Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levels |
| title_short | Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levels |
| title_sort | nitric oxide biosensor uncovers diminished ferrous iron dependency of cultured cells adapted to physiological oxygen levels |
| topic | Pericellular oxygen Hyperoxia Normoxia NO bioavailability geNOps Ferrous iron |
| url | http://www.sciencedirect.com/science/article/pii/S221323172200091X |
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