A Novel Strategy for Global Analysis of the Dynamic Thiol Redox Proteome

Nitroxidative stress in cells occurs mainly through the action of reactive nitrogen and oxygen species (RNOS) on protein thiol groups. Reactive nitrogen and oxygen species-mediated protein modifications are associated with pathophysiological states, but can also convey physiological signals. Identif...

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Bibliographic Details
Published in:Molecular & cellular proteomics 2012-09, Vol.11 (9), p.800-813
Main Authors: Martínez-Acedo, Pablo, Núñez, Estefanía, Gómez, Francisco J.Sánchez, Moreno, Margoth, Ramos, Elena, Izquierdo-Álvarez, Alicia, Miró-Casas, Elisabet, Mesa, Raquel, Rodriguez, Patricia, Martínez-Ruiz, Antonio, Dorado, David Garcia, Lamas, Santiago, Vázquez, Jesús
Format: Article
Language:English
Subjects:
Animals
Cell Hypoxia
Cells, Cultured
Endothelial Cells - metabolism
Humans
Isotope Labeling
Male
Mitochondria, Heart - metabolism
Myocardium - metabolism
Myocardium - pathology
Oxidation-Reduction
Oxidative Stress
Proteins - metabolism
Proteome - analysis
Proteomics
Rats
Rats, Sprague-Dawley
Reactive Nitrogen Species - analysis
Reactive Nitrogen Species - metabolism
Reactive Oxygen Species - analysis
Reactive Oxygen Species - metabolism
Reperfusion Injury - metabolism
Sulfhydryl Compounds - chemistry
Sulfhydryl Compounds - metabolism
Technological Innovation and Resources
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Summary:Nitroxidative stress in cells occurs mainly through the action of reactive nitrogen and oxygen species (RNOS) on protein thiol groups. Reactive nitrogen and oxygen species-mediated protein modifications are associated with pathophysiological states, but can also convey physiological signals. Identification of Cys residues that are modified by oxidative stimuli still poses technical challenges and these changes have never been statistically analyzed from a proteome-wide perspective. Here we show that GELSILOX, a method that combines a robust proteomics protocol with a new computational approach that analyzes variance at the peptide level, allows a simultaneous analysis of dynamic alterations in the redox state of Cys sites and of protein abundance. GELSILOX permits the characterization of the major endothelial redox targets of hydrogen peroxide in endothelial cells and reveals that hypoxia induces a significant increase in the status of oxidized thiols. GELSILOX also detected thiols that are redox-modified by ischemia-reperfusion in heart mitochondria and demonstrated that these alterations are abolished in ischemia-preconditioned animals.
ISSN:1535-9476
1535-9484
DOI:10.1074/mcp.M111.016469