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Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
Published on: June 21, 2021
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Improved integrative analysis of the thiol redox proteome using filter-aided sample preparation
Elena Bonzon-Kulichenko1, Emilio Camafeita1, Juan Antonio López1
1Cardiovascular Proteomics Laboratory, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Madrid, Spain; Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares (CIBERCV), Madrid, Spain.
Journal of Proteomics
|December 25, 2019
Summary
FASILOX, a new method, analyzes cysteine oxidation in proteins. It revealed men have a weaker antioxidant response than women, particularly in mitochondrial function, impacting obesity and diabetes.
Area of Science:
- Biochemistry
- Cellular signaling
- Metabolomics
Background:
- Protein cysteine oxidation is crucial in cell signaling and disease.
- Previous GELSILOX method allowed large-scale redox state analysis.
- Obesity and diabetes involve altered cellular redox states.
Purpose of the Study:
- Introduce FASILOX, an improved method for analyzing cysteine oxidation.
- Enhance peptide recovery, sensitivity, and workflow efficiency.
- Investigate sex differences in cysteine oxidation in obesity and diabetes.
Main Methods:
- Developed FASILOX, an advanced in-gel proteomic technique.
- Applied FASILOX to mitochondria from obese, diabetic patients' adipose tissue.
- Analyzed dynamic alterations in cysteine redox states and protein abundance.
Main Results:
- FASILOX demonstrated increased peptide recovery and sensitivity.
- A sexually dimorphic pattern of cysteine oxidation was observed.
- Oxidative alterations primarily affected mitochondrial oxidative phosphorylation complexes.
Conclusions:
- FASILOX offers a streamlined and sensitive approach for redox proteomics.
- Men exhibit a less efficient antioxidant response compared to women.
- Cysteine oxidation differences may contribute to pathophysiology in obesity and diabetes.
