Journal
REDOX BIOLOGY
Volume 2, Issue -, Pages 955-962Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.redox.2014.08.001
Keywords
Hydrogen peroxide; Peroxiredoxin; Kinetics; HyPer; Quantitative redox biology
Categories
Funding
- NSF Graduate Research Fellowship [2009085738]
- Burroughs Wellcome Fund Career Award at the Scientific Interface [1005762]
- Joseph R. Mares Professorship in Chemical Engineering
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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 horn 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. (C) 2014 The Authors. Published by Elsevier B.V.
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