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Quantifying intracellular hydrogen peroxide perturbations in terms of concentration

Author(s)
Huang, Beijing K.; Sikes, Hadley
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Abstract
Molecular level, mechanistic understanding of the roles of reactive oxygen species (ROS) in a variety of pathological conditions is hindered by the difficulties associated with determining the concentration of various ROS species. Here, we present an approach that converts fold-change in the signal from an intracellular sensor of hydrogen peroxide into changes in absolute concentration. The method uses extracellular additions of peroxide and an improved biochemical measurement of the gradient between extracellular and intracellular peroxide concentrations to calibrate the intracellular sensor. By measuring peroxiredoxin activity, we found that this gradient is 650-fold rather than the 7–10-fold that is widely cited. The resulting calibration is important for understanding the mass-action kinetics of complex networks of redox reactions, and it enables meaningful characterization and comparison of outputs from endogenous peroxide generating tools and therapeutics across studies.
Date issued
2014-08
URI
http://hdl.handle.net/1721.1/101232
Department
Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Chemical Engineering
Journal
Redox Biology
Publisher
Elsevier
Citation
Huang, Beijing K., and Hadley D. Sikes. “Quantifying Intracellular Hydrogen Peroxide Perturbations in Terms of Concentration.” Redox Biology 2 (2014): 955–962.
Version: Final published version
ISSN
22132317

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