Genetically Encoded Biosensor HyPer as a Tool for Quantification of Intracellular Hydrogen Peroxide Concentrations

This mini-reviewsystematizes information on methods for quantitative assessment of intracellular hydrogenperoxide concentration based on the use of a genetically encodedperoxide sensor HyPer. Two approaches are being considered: 1) calibrationof the biosensor using exogenous hydrogen peroxide, based on assessingthe rate of peroxide penetration into cells and intracellular peroxidaseactivity; 2) direct determination of the intracellular peroxide content, basedon measuring the level of oxidation of the biosensor, theoxidation reaction constant and the reduction reaction constant of HyPerin the cells. The use of these methods makes itpossible to solve a wide range of tasks in cellularredox biology—to determine the range of physiological anddamaging concentrations of hydrogen peroxide in cells, to evaluate theeffectiveness of the antioxidant defense system in various cellular compartmentsunder conditions of oxidative stress, to determine the contribution ofvarious enzymatic systems to the peroxidase activity of cells, andto characterize antioxidant defense systems in various biological contexts (inthe process of cellular senescence, differentiation, reprogramming, during the developmentof pathologies). The described methods can be adapted for othergenetically encoded hydrogen peroxide biosensors.