Molecular Biology, Genetics and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, Turkey.
King'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.
Redox Biol. 2022 Jul;53:102319. doi: 10.1016/j.redox.2022.102319. Epub 2022 Apr 30.
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 O) or physiological normoxia (5 kPa O). 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.
铁是细胞代谢和信号传递所必需的金属，但过量也会产生不良影响。缺铁的生理后果已得到充分证实，然而，铁补充与培养细胞周围氧水平及其对金属蛋白酶下游影响之间的关系尚未得到充分探索。本研究利用培养的 HEK293T 上皮细胞和 EA.hy926 内皮细胞中的金属蛋白酶 geNOps，测试适应标准室内空气（18 kPa O）或生理常氧（5 kPa O）的细胞中的铁依赖性。我们表明，培养中的细胞需要铁补充来激活金属蛋白酶 geNOps，并首次证明适应生理常氧的细胞比适应高氧的细胞需要的铁明显更少。本研究确立了在体重现氧水平的重要作用，并揭示了在标准细胞培养条件下需要亚铁补充以实现 geNOps 功能的以前未被认识到的需求。
