Redox Proteomics Service

Redox proteomics is a field of study focused on the redox state and dynamic changes of proteins, aiming to analyze redox modifications (such as nitration, carbonylation, and thiol modifications) and their regulatory roles in protein function, signaling pathways, and diseases. Oxidative modifications induced by reactive oxygen species (ROS) and reactive nitrogen species (RNS) can be both the result of oxidative stress and critical elements of signal regulation. By quantitatively analyzing the oxidative states of proteins, redox proteomics provides new insights into protein activity, protein-protein interactions, and their spatiotemporal regulation in relation to oxidative stress, laying the foundation for exploring molecular mechanisms of diseases and potential therapeutic targets.

 

Redox proteomics research involves complex analytical workflows that combine various techniques and instruments to achieve comprehensive detection of oxidative modifications. Immunochemical methods (such as ELISA, Western Blot, and immunohistochemistry) are suitable for high-throughput qualitative or semi-quantitative analysis of specific modifications but are limited in sensitivity. Chromatographic techniques (such as HPLC-MS and GC-MS) are widely used for the high-selectivity detection of nitration and carbonylation modifications, with HPLC-MS being the gold standard for biological sample analysis. Mass spectrometry (such as LC-MS/MS) serves as the core tool for identifying and quantifying oxidative modifications. By integrating TMT labeling, Switch Assay, or RAC techniques, redox proteomics enables high-throughput and precise analysis of modification sites. These techniques collectively establish a comprehensive research platform capable of uncovering dynamic changes in oxidative modifications, analyzing the remodeling of redox networks, and identifying disease-related oxidative biomarkers, providing powerful tools for disease diagnosis and therapy.

 

Table 1. Main Applied Methods in the Study of Redox Proteomics



Cadenas-Garrido, P. et al. Antioxidants (Basel). 2024.

 

Services at MtoZ Biolabs

MtoZ Biolabs provides cutting-edge redox proteomics service designed to accurately detect and quantify protein redox modifications using high-sensitivity mass spectrometry platforms like LC-MS/MS. Covering modifications such as nitration, carbonylation, and thiol oxidation, our redox proteomics service reveals dynamic changes in protein oxidation states, offering insights into their roles in cellular signaling and disease progression. By combining high-throughput analysis with tailored solutions, we empower researchers to uncover oxidative stress mechanisms and identify biomarkers, delivering valuable support for advancing disease research and therapeutic development. If you are interested in our service, please contact us freely.

 



Day, NJ. et al. Antioxidants (Basel). 2021.

Figure 1. Workflow of Different Quantitative Redox Proteomics Approaches

Service Advantages

1. High-Throughput and Precise Detection 

MtoZ Biolabs utilizes advanced mass spectrometry platforms (such as Orbitrap and TIMS-TOF), combined with TMT labeling and RAC techniques, to achieve high-throughput and precise identification and quantification of oxidative modification sites, including carbonylation, nitration, and sulfonation, enabling comprehensive analysis of protein oxidation states.

 

2. Dynamic Analysis  

MtoZ Biolabs' redox proteomics service leverages quantitative redox proteomics techniques to dynamically track real-time changes in oxidative modifications, aiding in the understanding of oxidative stress, redox network remodeling, and their roles in disease progression.

 

3. Customized Research Solutions  

Through our redox proteomics service, MtoZ Biolabs helps clients delve into the critical roles of oxidative modifications in diseases, providing customized research solutions to identify disease-related oxidative biomarkers and potential therapeutic targets, supporting precision medicine research.

 

Case Study

Case 1: This study uses chemical proteomics to identify new targets of cysteine sulfinic acid reductase (CSAD), revealing the enzyme's critical role in regulating the cellular redox state. Our redox proteomics service can analyze the redox modifications of the enzyme and its targets, helping to explore their roles in cellular functions.

 



Akter, S. et al. Nat Chem Biol. 2018.

Figure 2. Characterizing S-Sulfinylation Sites Using Redox Proteomics and Chemoproteomics Approaches

 

Case 2: The study highlights how redox regulation varies across tissues during aging, identifying key proteins and pathways affected by oxidative modifications. Using advanced redox profiling, the research reveals age-associated shifts in redox-sensitive cysteine residues and their impact on cellular functions. Redox proteomics service enables the precise quantification of such oxidative modifications, mapping tissue-specific redox changes and identifying potential biomarkers of aging. This service provides powerful tools to study the molecular mechanisms of aging and oxidative stress.

 



Xiao, H. et al. Cell. 2020.

Figure 3. Tissue-Specific and Age-Dependent Cysteine Oxidation Changes Analyzed by Quantitative Redox Proteomics

 

Applications

1. Gaining New Insights into Disease through Redox Proteomics

Antioxidant defense maintains redox homeostasis and regulates baseline levels of reactive oxygen and nitrogen species (ROS and RNS). Oxidative stress (OS), characterized by insufficient antioxidant defense or elevated ROS/RNS, leads to biomolecular modifications, primarily affecting proteins. Proteomics provides dynamic insights into protein expression, modifications, localization, and interactions, revealing key oxidative processes like carbonylation, glycation, and lipid peroxidation. Reactive enals (e.g., 4-hydroxy-2-nonenal) modify cysteine, lysine, and histidine residues through Michael addition, while tyrosine undergoes nitration and cysteine nitrosylation. Oxidative and nitrosative stress, especially in the brain, is linked to neurodegenerative diseases due to high metabolic demand. Redox proteomics methods for detecting, identifying, and quantifying oxidative modifications are crucial for understanding these mechanisms. Redox proteomics service leverages advanced LC-MS/MS to identify oxidative modifications, enabling precise analysis of protein oxidation states, critical in studying disease progression and oxidative stress mechanisms.

 



Cadenas-Garrido, P. et al. Antioxidants (Basel). 2024.

Figure 4. Redox Proteomics: Impact on Human Health and New Insights

 

FAQ

Q1: In redox proteomics research, how can protein modifications caused by physiological redox signaling regulation be distinguished from pathological modifications induced by oxidative stress?

Answer: In redox proteomics research, distinguishing between physiological redox regulation and pathological oxidative stress-related protein modifications requires dynamic analysis and high-precision quantification techniques. Physiological redox regulation is typically reversible, such as cysteine oxidation, while pathological oxidative stress often leads to irreversible modifications like protein carbonylation or lipid peroxidation. Using redox proteomics service with high-resolution mass spectrometry, specific modifications (e.g., nitrated tyrosine or 4-hydroxy-2-nonenal-modified cysteine) can be quantitatively analyzed, and isotopic labeling methods can compare differences between normal and stress conditions. Additionally, integrating functional cellular studies to validate whether modifications are physiological or pathological is crucial. The redox proteomics service provides a highly sensitive platform to uncover the specific roles of redox modifications in pathological mechanisms such as neurodegenerative diseases.

 

Deliverables

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. Relevant Liquid Chromatography and Mass Spectrometry Parameters

4. The Detailed Information of Redox Proteomics

5. Mass Spectrometry Image

6. Raw Data