Nitrosylation Analysis Service
Nitrosylation is a reversible post-translational modification that regulates protein activity, localization, and signaling, playing vital roles in both physiological and pathological processes. MtoZ Biolabs provides comprehensive Nitrosylation Analysis Services tailored to different research needs, encompassing both targeted protein nitrosylation analysis and nitrosylation proteomics. By integrating advanced enrichment techniques, high-resolution LC-MS/MS, and bioinformatics interpretation, we offer accurate identification, quantification, and biological insight into protein nitrosylation dynamics.
1. Target Protein Nitrosylation Analysis
This service focuses on the precise detection and quantification of nitrosylation on a specific protein of interest. Through selective enrichment strategies such as the biotin-switch assay or resin-assisted capture combined with high-resolution mass spectrometry, we accurately identify modification sites and assess nitrosylation levels under various biological conditions. This targeted approach enables detailed investigation of nitrosylation’s regulatory role in the function and signaling of a given protein.
2. Nitrosylation Proteomics
For large-scale studies, MtoZ Biolabs offers proteome-wide nitrosylation profiling. Utilizing optimized enrichment protocols and advanced LC-MS/MS platforms, this service systematically characterizes nitrosylation events across thousands of proteins. The resulting datasets reveal global modification patterns, identify affected pathways, and support comparative analyses between disease and control samples. Our comprehensive bioinformatics analysis provides functional annotation, pathway mapping, and quantitative comparisons, offering a panoramic view of nitrosylation regulation in complex biological systems.
What is Nitrosylation?
Nitrosylation is one of the key post-translational modifications mediated by nitric oxide (NO) signaling. Endogenously produced by nitric oxide synthases (NOS), NO can directly react with cysteine side chains of proteins, forming S-nitrosylation (SNO). This modification is reversible and can be propagated through transnitrosylation, in which the nitrosyl group is transferred between cysteine residues of different proteins. It can also be reversed by the cellular redox system, particularly through S-nitrosoglutathione reductase (GSNO-R), which regulates the turnover of S-nitrosoglutathione (GSNO) in cooperation with glutathione and glutaredoxins. Beyond cysteine modification, NO-derived reactive intermediates can also induce irreversible modifications, such as 3-nitrotyrosine, formed by the interaction of NO with superoxide and peroxynitrite.
Spiers, J. G. et al. Channels (Austin). 2021.
Figure 1. Post-Translational Modifications Mediated by NO Signaling
Early discoveries of nitrosylated hemoglobin and caspase proteins highlighted their roles in oxygen transport and apoptosis regulation. Subsequent studies have revealed nitrosylation to be widespread across different protein families, ranging from metabolic enzymes to transcription factors and signaling receptors. Functionally, nitrosylation regulates vascular tone, neurotransmission, immune defense, and cellular adaptation to oxidative environments. Dysregulated nitrosylation has been implicated in cardiovascular diseases, cancer, neurodegenerative disorders such as Alzheimer's and Parkinson's disease, and chronic inflammatory conditions. Precise nitrosylation analysis is essential for mapping modification sites, quantifying changes under different biological states, and uncovering disease-related mechanisms that can guide biomarker discovery and therapeutic development.
Analysis Workflow
Sample Submission Suggestions
🔸Storage and Shipping: Samples should be flash-frozen in liquid nitrogen and shipped on dry ice.
🔸Replicates: At least 3 biological replicates are recommended for statistical reliability.
*Note: If you have special sample types or require additional guidance, please contact us for personalized support before sample preparation.
Service Advantages
✔️State-of-the-Art Mass Spectrometry ensuring high sensitivity and accuracy.
✔️Specialized Enrichment Techniques for confident identification of labile S-nitrosylated peptides.
✔️Comprehensive Options from targeted assays to global proteomics analysis.
✔️Experienced Scientific Team with expertise in PTM analysis and bioinformatics.
✔️High Data Quality with stringent quality control throughout the workflow.
✔️Customizable Solutions tailored to specific research objectives.
✔️Rapid Turnaround enabling efficient progress from sample submission to results.
Applications
Nitrosylation has emerged as a central regulator of diverse biological processes, and comprehensive Nitrosylation Analysis Service provides valuable insights across multiple research areas.
1. Redox Regulation
Nitrosylation is a critical component of redox signaling networks, allowing cells to sense and adapt to changes in oxidative environments. By analyzing nitrosylation dynamics, researchers can better understand how proteins integrate redox signals to regulate cellular homeostasis and stress responses.
2. Signal Transduction
As a reversible modification, nitrosylation modulates the activity of enzymes, receptors, and transcription factors, influencing pathways involved in vascular function, neuronal communication, and immune regulation. Accurate mapping of nitrosylation events supports the dissection of complex signaling cascades and the identification of novel regulatory checkpoints.
3. Disease Mechanisms
Aberrant nitrosylation is implicated in a wide spectrum of diseases, including cardiovascular disorders, cancer, and neurodegenerative conditions. Nitrosylation analysis enables the identification of disease-associated protein modifications, supports the search for reliable biomarkers, and aids in the discovery of potential therapeutic targets. By providing quantitative and site-specific information, this service contributes to translational studies bridging fundamental biology and clinical research.
Deliverables
1. Comprehensive Experimental Details
2. Materials, Instruments, and Methods
3. The Detailed Information of Nitrosylation Analysis
4. Mass Spectrometry Image
5. Bioinformatics Analysis
6. Raw Data
If you are interested in our Nitrosylation Analysis Service, please feel free to contact us. Our technical specialists are available to provide a free business assessment.