Related Experiment Video
Updated: Jun 22, 2026
11:49
Quantitative Proteomics Workflow using Multiple Reaction Monitoring Based Detection of Proteins from Human Brain Tissue
Published on: August 28, 2021
Proteomics identification of oxidatively modified proteins in brain
Rukhsana Sultana1, Marzia Perluigi, D Allan Butterfield
1Department of Chemistry, Sanders-Brown Center on Aging, University of Kentucky, Lexington, KY 40506-0055, USA.
Methods in Molecular Biology (Clifton, N.J.)
|June 23, 2009
Summary
This study introduces redox proteomics to identify protein oxidation in neurodegenerative diseases. The method detects carbonylated and HNE-bound proteins, crucial for understanding oxidative stress.
Area of Science:
- Biochemistry
- Neuroscience
- Proteomics
Background:
- Reactive oxygen species (ROS) contribute to neurodegenerative diseases via oxidative damage.
- Protein oxidation disrupts cellular signaling, structure, and metabolism, exacerbating oxidative stress.
Purpose of the Study:
- To describe the protocol for redox proteomics.
- To focus on identifying carbonylated and 4-hydroxy-2-trans-nonenal (HNE)-bound proteins.
Main Methods:
- Redox proteomics protocol detailed for identifying protein carbonyls and HNE adducts.
- Protein carbonyls derivatized with DNPH, followed by 2D separation and Western blot detection.
- HNE-bound proteins detected similarly, without derivatization.
Main Results:
- The described redox proteomics approach enables identification of specific protein modifications.
- Carbonylated and HNE-bound proteins are successfully detected using 2D Western blot.
- Mass spectrometry and database interrogation used for final protein identification.
Conclusions:
- Redox proteomics is a valuable tool for studying post-translational modifications in neurodegenerative diseases.
- The protocol facilitates the identification of key oxidative damage markers.
- Understanding these modifications aids in elucidating disease pathophysiology.
