Quantifying intracellular hydrogen peroxide perturbations in terms of concentration
| Autor: | Hadley D. Sikes, Beijing K. Huang |
|---|---|
| Přispěvatelé: | Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Chemical Engineering, Huang, Beijing K., Sikes, Hadley |
| Rok vydání: | 2014 |
| Předmět: |
Clinical Biochemistry
Biosensing Techniques Quantitative redox biology Biochemistry Redox Peroxide chemistry.chemical_compound Extracellular Humans Hydrogen peroxide lcsh:QH301-705.5 HyPer chemistry.chemical_classification lcsh:R5-920 Reactive oxygen species Peroxiredoxin activity Organic Chemistry Peroxiredoxin Kinetics lcsh:Biology (General) chemistry lcsh:Medicine (General) Intracellular HeLa Cells Research Paper |
| Zdroj: | Redox Biology, Vol 2, Iss C, Pp 955-962 (2014) Redox Biology Elsevier Open Access |
| ISSN: | 2213-2317 |
| DOI: | 10.1016/j.redox.2014.08.001 |
| Popis: | 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. National Science Foundation (U.S.). Graduate Research Fellowship (2009085738) Burroughs Wellcome Fund (Career Award at the Scientific Interface 1005762) |
| Databáze: | OpenAIRE |
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