Protein-thiol oxidation, from single proteins to proteome-wide analyses

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Protein-thiol oxidation is crucial for cellular functions including catalysis, folding, trafficking, and sensing reactive oxygen and nitrogen species (ROS/RNS).
• Dysregulated protein-thiol oxidation can result from ROS/RNS toxicity, necessitating precise monitoring methods.
• Understanding the in vivo redox state of cysteine residues is vital for cellular health and disease research.

Purpose of the Study:

• To develop accurate and robust biochemical methods for monitoring the in vivo redox state of cysteine residues.
• To enable the discovery of novel proteins and pathways involved in cellular redox regulation.

Main Methods:

• Developed biochemical approaches to trap cysteine residues in their in vivo redox state under acidic conditions.
• Utilized differential labeling of reduced versus oxidized cysteine residues with thiol-specific reagents.
• Applied proteome-wide strategies to analyze cytoplasmic oxidized protein thiols.

Main Results:

• Successfully identified eukaryotic peroxide receptors and novel ROS-scavenging enzymes.
• Characterized the repertoire of cytoplasmic oxidized protein thiols.
• Established the roles of thioredoxin and glutathione pathways in eukaryotic cytoplasmic thiol-redox control.

Conclusions:

• The developed methods provide a powerful tool for studying protein-thiol oxidation in vivo.
• These techniques have advanced the understanding of cellular redox sensing, signaling, and regulation.
• The findings highlight the importance of redox control pathways in maintaining cellular homeostasis.