Cysteine, a sulfur-containing amino acid, plays a pivotal role in cellular redox signaling. The burgeoning field of Cysteine-Redoxome Proteomics focuses on deciphering the intricate network of redox modifications involving cysteine residues within proteins.
Cysteine-redoxome proteomics analysis is important because it sheds light on the complex and dynamic cellular mechanisms involved in redox control. In many physiological and pathological situations, reactive oxygen species (ROS) and reactive nitrogen species (RNS) play a key role in redox signaling. Because cysteine residues in proteins are particularly susceptible to redox alterations, researchers can better understand the functional effects of these modifications by examining the Cysteine-redoxome proteomics. Deciphering the intricacies of redox signaling in health and illness necessitates the analysis of cystine-redoxome proteomics. It advances not just our knowledge of basic biological functions but also has applications in personalized medicine and treatment development.
Analytical Methods of Cysteine-Redoxome Proteomics
- Mass Spectrometry. Mass spectrometry (MS) stands at the forefront of analytical methods employed in cysteine-redoxome proteomics. Its ability to precisely identify and quantify redox modifications on cysteine residues has revolutionized our understanding of cellular redox dynamics. Techniques such as tandem mass spectrometry (MS/MS) enable the characterization of specific redox states, offering a detailed view of this landscape.
- Redox-Sensitive Probes. Intriguing advancements involve the use of redox-sensitive probes that selectively react with modified cysteine residues. Fluorescent tags coupled with these probes facilitate real-time imaging of redox changes within live cells. This approach not only provides spatial information but also allows for the temporal tracking of redox events, unraveling the dynamic nature of cellular redox signaling.
- Bioinformatics. The complexity of cysteine-redoxome proteomics generates vast datasets that necessitate sophisticated bioinformatic tools for interpretation. Integrative approaches, including machine learning algorithms, aid in the identification of redox-sensitive proteins and the prediction of potential redox modification sites. Bioinformatics not only accelerates data analysis but also guides subsequent experimental design.
Application of Cysteine-Redoxome Proteomics
- Neurodegenerative Diseases. Cysteine-redoxome proteomics has emerged as a powerful tool in unraveling the molecular underpinnings of neurodegenerative diseases. By profiling redox modifications in key proteins associated with conditions like Alzheimer's and Parkinson's, researchers gain insights into the role of oxidative stress in disease progression. This knowledge opens avenues for targeted therapeutic interventions.
- Cancer Biology. The dysregulation of redox signaling is a hallmark of cancer cells. Cysteine-redoxome proteomics contributes to our understanding of how altered redox states drive oncogenic processes. Identifying redox-sensitive proteins in cancer cells offers potential targets for the development of novel anticancer therapies, capitalizing on the vulnerabilities induced by redox imbalance.
- Drug Development. Harnessing the information garnered from cysteine-redoxome proteomics, drug development endeavors now incorporate a redox-sensitive perspective. Designing therapeutics that respond to the redox status of specific proteins allows for a more nuanced and targeted approach. This strategy holds promise in mitigating off-target effects and enhancing the efficacy of pharmaceutical interventions.
- Redoxome Dynamics in Aging. As cells age, redox homeostasis undergoes significant alterations. Cysteine-redoxome proteomics contributes to the understanding of how redox modifications evolve with aging, shedding light on the molecular mechanisms that underlie age-related pathologies. Unraveling the redoxome dynamics in aging provides a foundation for developing interventions to promote healthy aging.
- Environmental Stress Responses. Cells encounter various environmental stresses that impact their redox status. Cysteine-redoxome proteomics enables researchers to delineate the redox changes induced by stressors such as oxidative, nitrosative, and reductive stress. Understanding how cells adapt to environmental challenges at the redox level informs strategies for enhancing stress resilience in various biological systems.
- Systems Biology and Network Analysis. Comprehensive comprehension of biological systems is enhanced by the analysis of the proteomics of the cysteine-redoxome. A systems biology approach is made possible by integration with other omics data, including transcriptomics, metabolomics, and genomes. Building networks and pathways that provide a more realistic depiction of biological processes and their regulation is made possible by this all-encompassing viewpoint.
Our Service Advantages
Cysteine-redoxome proteomics stands as a transformative discipline in the realm of cellular redox signaling. Creative Proteomics provides advanced analytical methods to help researchers advance the understanding of the cysteine redoxome. As we continue to unravel the complexities of the cysteine redoxome, the potential for novel therapeutic targets and redox-responsive interventions becomes increasingly promising.
Global profiling of distinct cysteine redox forms reveals wide-ranging redox regulation in C.elegans
Journal: Nat Commun
Published: 2021
Background
Redox modification of cysteine plays an important role as a part of protein post-translational modification. In this thesis, the model organism Cryptomeria japonica was used as a carrier to explore the multiple redox forms of cysteine, revealing that cysteine redox can regulate various biological processes and pathways in the nematode, and finding that proteins upstream and downstream of the p38 MAPK pathway can mediate Cys-dependent antioxidant response and natural immune regulation.
Results
The authors applied the technique of quantification of protein sulfhydryl reactivity to wild-type L4 stage nematodes. Treatment with high (100 uM) and low (10 uM) doses of IPM probes detected reactivity at a total of 5258 cysteine sites, greatly extending the coverage of the cysteine proteome. The analysis suggests that there may be significant differences in the reactivity of cysteines in the same protein, and that the reactivity of cysteines can be used as an effective predictor of their function (Figure 1).
Figure 1
The authors used a redox proteomics approach based on "click chemistry" probes to accurately quantify the major cysteine redox forms in complex proteomes. The authors treated wild-type nematodes with 5 mM H2O2 for 5 min and labeled them with different probes. The results showed that 5453 Cys-SH sites were localized on 2864 proteins, 1521 Cys-SOH sites on 1049 proteins and 82 Cys-SO2H sites on 72 proteins. And more than 1500 protein Cys sites were found to be significantly changed after exogenous low concentration H2O2 treatment (Figure 2).
Figure 2
The authors found redox-modified proteins localized in different tissues and organs, which were subjected to GO and KEGG analysis. Bioinformatics analysis revealed that these oxidation-sensitive proteins are involved in numerous biological pathways. The focus was on whether SEK-1 and PMK-1 play a redox-regulatory role in p38 MAPK-mediated stress response and pathogen resistance (Figure 3). The authors constructed a point mutant nematode model using Cas9/CRISPR gene editing and found that both C173 of PMK-1 and C213 of SEK-1 are involved in oxidative stress-mediated activation of the p38MAPK pathway, and that this is critical for nematode resistance to pathogens (Figure 4).
Figure 3
Figure 4
Conclusion
Cellular redox states and cysteine modifications are common during organismal aging, and cysteine redox profiles and redox state driver proteins may also be potential biomakers under physiological and pathological conditions.The authors' dataset of quantitative, site-centered mapping of intrinsic cysteine responsiveness and peroxide-sensitive cysteine redox forms greatly expands the the scope and biological role of cysteine oxidation in nematodes and provide a molecular basis for deciphering the complex redox signaling network in model organisms.
Reference
- Meng J, Fu L, Liu K, et al,. Global profiling of distinct cysteine redox forms reveals wide-ranging redox regulation in C. elegans. Nat Commun. 2021 Mar 3;12(1):1415.
