Which Antioxidant System Shapes Intracellular H ₂ O ₂ Gradients?

Abstract

Cellular antioxidant systems control the levels of hydrogen peroxide (H₂O₂) within cells. Multiple theoretical models exist that predict the diffusion properties of H₂O₂ depending on the rate of H₂O₂ generation and amount and reaction rates of antioxidant machinery components. Despite these theoretical predictions, it has remained unknown how antioxidant systems shape intracellular H₂O₂ gradients. The relative role of thioredoxin (Trx) and glutathione systems in H₂O₂ pattern formation and maintenance is another disputed question. Here, we visualized cellular antioxidant activity and H₂O₂ gradients formation by exploiting chemogenetic approaches to generate compartmentalized intracellular H₂O₂ and using the H₂O₂ biosensor HyPer to analyze the resulting H₂O₂ distribution in specific subcellular compartments. Using human HeLa cells as a model system, we propose that the Trx system, but not the glutathione system, regulates intracellular H₂O₂ gradients. <i>Antioxid. Redox Signal.</i> 31, 664-670.