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Published on: June 21, 2021
A simple method to systematically study oxidatively modified proteins in biological samples and its applications
Byoung-Joon Song1, Soo-Kyung Suh, Kwan-Hoon Moon
1Laboratory of Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism, Bethesda, Maryland, USA.
Abstract:
Increased oxidative stress with elevated levels of reactive oxygen/nitrogen species (ROS/RNS) plays an important role in the pathophysiology of many disease states. Increased ROS/RNS can modulate the cellular macromolecules of DNA, lipids, and proteins, negatively affecting their normal functions. Numerous reports have described the properties and implications of oxidized DNA and lipids. However, oxidative modifications of proteins were not fully studied partially due to the requirement for specific reagents, the lack of methods to detect, purify, and identify oxidatively modified proteins, and the relatively late development of highly sensitive analytical instruments. This chapter describes the detailed procedure for systematically identifying oxidative-modified proteins in biological samples. Applications and other suggestions to this method are also described to understand the functional roles of oxidatively modified proteins in promoting endoplasmic reticulum (ER) stress and mitochondrial dysfunction, which ultimately contribute to organ damage.
Insights
Oxidative stress from reactive oxygen/nitrogen species (ROS/RNS) impacts disease. This study details a method to identify oxidized proteins, crucial for understanding their role in ER stress and organ damage.
Area of Science:
- Biochemistry
- Molecular Biology
- Pathophysiology
Background:
- Oxidative stress, characterized by elevated reactive oxygen/nitrogen species (ROS/RNS), is implicated in numerous diseases.
- While oxidized DNA and lipids are well-studied, oxidative modifications in proteins remain less understood due to methodological challenges.
- Challenges include the need for specific reagents, difficulties in detection and purification, and limitations of early analytical instruments.
Purpose of the Study:
- To present a detailed procedure for systematically identifying oxidatively modified proteins in biological samples.
- To highlight the functional significance of these modifications in disease pathogenesis.
- To provide insights into the role of oxidized proteins in endoplasmic reticulum (ER) stress and mitochondrial dysfunction.
Main Methods:
- Detailed procedural description for the systematic identification of oxidatively modified proteins.
- Utilizes advanced analytical techniques for sensitive detection and purification.
- Focuses on biological samples to ensure relevance to disease states.
Main Results:
- A comprehensive method for identifying oxidatively modified proteins is established.
- The method facilitates understanding the functional consequences of protein oxidation.
- Demonstrates the link between oxidized proteins and cellular dysfunction.
Conclusions:
- The developed method enables systematic identification of oxidatively modified proteins.
- Understanding protein oxidation is key to elucidating disease mechanisms.
- This approach aids in studying the contribution of oxidized proteins to ER stress, mitochondrial dysfunction, and organ damage.
