GO:0035606 peptidyl-cysteine S-trans-nitrosylation: Protein Modification Mechanism, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0035606 peptidyl-cysteine S-trans-nitrosylation is defined as the transfer of a nitric oxide (NO) group from one cysteine residue to another.
• This process is a post-translational modification that can alter protein function, localization, and activity, and is distinct from other cysteine oxidation events.
• Enzymes such as formylglycine-generating enzyme (FGE) and related copper-dependent oxidases provide mechanistic insights into cysteine oxidation chemistry relevant to transnitrosylation.
• Cysteine-to-cysteine transnitrosylation is implicated in redox signaling and may influence cellular responses in diseases such as diabetes and periodontitis.
• NADPH oxidase-dependent redox homeostasis in fungal pathogens highlights the broader biological importance of cysteine-based redox regulation.
• CRISPR-based models (knockout, point mutation, knock-in, overexpression) enable causal dissection of genes involved in peptidyl-cysteine S-trans-nitrosylation.
Description
Peptidyl-cysteine S-trans-nitrosylation (GO:0035606) is a biological process defined as the transfer of a nitric oxide (NO) group from one cysteine residue to another. This reaction represents a specific type of post-translational modification where a nitroso group is passed between protein cysteine thiols, potentially altering protein function and contributing to redox signaling networks. Understanding this process is critical because cysteine residues are highly reactive and susceptible to various oxidative modifications, and their regulation impacts diverse cellular pathways. Researchers study peptidyl-cysteine S-trans-nitrosylation to uncover how NO signaling is propagated and controlled at the protein level, and how dysregulation may contribute to disease. The reaction is chemically related to other cysteine oxidation events catalyzed by copper-dependent enzymes such as formylglycine-generating enzyme (FGE), which has been structurally and mechanistically characterized. These studies provide a framework for understanding the enzymatic and non-enzymatic mechanisms that could underlie transnitrosylation. Furthermore, proteomic analyses in clinical samples, such as saliva from patients with type 2 diabetes and periodontitis, have identified changes in proteins that may be subject to redox modifications, underscoring the potential disease relevance of cysteine-based modifications. In fungal pathogens, NADPH oxidase is crucial for cellular redox homeostasis, illustrating the broad importance of redox regulation across organisms. Thus, GO:0035606 sits at the intersection of nitric oxide biology, redox chemistry, and post-translational modification research, with implications for both basic science and translational medicine.
peptidyl-cysteine S-trans-nitrosylation At A Glance
| GO ID | GO:0035606 |
|---|---|
| GO term | peptidyl-cysteine S-trans-nitrosylation |
| Ontology | biological_process |
| Synonym | cysteine to cysteine nitrosylation; cysteine-to-cysteine transnitrosylation; protein-to-protein transnitrosylation; S-transnitrosylation |
| Major function | Transfer of a nitric oxide (NO) group from one cysteine residue to another |
| Related chemistry | Cysteine oxidation and nitrosylation chemistry studied in copper-dependent enzymes |
| Disease relevance | Implicated in redox-related pathologies such as diabetes and periodontitis |
| Organismal context | Redox homeostasis in fungal pathogens involves NADPH oxidase |
What Is GO:0035606?
Peptidyl-cysteine S-trans-nitrosylation is the transfer of a nitric oxide (NO) group from one cysteine residue to another. In this process, a nitroso group is passed from a donor cysteine (often on one protein) to an acceptor cysteine (on another protein or the same protein), resulting in S-nitrosylation of the acceptor cysteine. This definition distinguishes it from other cysteine modifications such as oxidation to sulfenic acid or disulfide formation, as it specifically involves NO group transfer.
Why Is peptidyl-cysteine S-trans-nitrosylation Important in Cell Biology?
Peptidyl-cysteine S-trans-nitrosylation is important because it represents a mechanism for propagating nitric oxide signaling through protein cysteine residues, thereby influencing protein activity, interactions, and stability. This process is part of the broader redox regulatory network that controls cellular responses to stress and infection. Dysregulation of cysteine modifications has been linked to human diseases, including metabolic and inflammatory conditions, as suggested by proteomic studies in diabetes and periodontitis. Understanding the enzymes and chemical mechanisms that facilitate cysteine oxidation, such as those involving formylglycine-generating enzyme, provides a foundation for targeting these pathways.
• Provides a mechanism for NO group transfer between cysteines, expanding the repertoire of post-translational modifications.
• Impacts protein function and redox signaling, with potential roles in cellular stress responses.
• Chemically related to cysteine oxidation reactions catalyzed by copper-dependent enzymes like FGE.
• May contribute to disease processes such as diabetes and periodontitis, as indicated by proteomic changes.
• Relevant to host-pathogen interactions, as redox homeostasis in fungi depends on NADPH oxidase.
• Offers targets for therapeutic intervention in diseases involving aberrant NO signaling.
• Enables researchers to study protein-protein interactions mediated by NO transfer.
• Connects to broader redox biology through shared cysteine chemistry.
• Can be investigated using CRISPR models to dissect gene function in redox pathways.
• Highlights the importance of cysteine residues as regulatory switches in proteins.
What Happens During peptidyl-cysteine S-trans-nitrosylation?
Donor cysteine activation
In simple terms: A cysteine on a donor protein gets modified with a nitric oxide group.
The process begins with a donor cysteine residue that carries a nitric oxide (NO) group, often formed via prior S-nitrosylation. This activated cysteine serves as the source for transfer. The chemical nature of this step is related to cysteine oxidation chemistry observed in enzymes like formylglycine-generating enzyme, where copper-dependent activation of oxygen leads to cysteine modification.
NO group transfer
In simple terms: The nitric oxide group is handed directly from one cysteine to another.
The NO group is transferred from the donor cysteine to an acceptor cysteine residue, which can be on the same or a different protein. This transnitrosylation reaction results in S-nitrosylation of the acceptor cysteine. The transfer may be facilitated by proximity and the redox environment, and is distinct from free NO diffusion.
Acceptor cysteine modification
In simple terms: The receiving cysteine becomes nitrosylated, which can change how the protein works.
Upon receiving the NO group, the acceptor cysteine undergoes S-nitrosylation, a modification that can alter protein conformation, activity, or interactions. This modification is reversible and part of cellular redox regulation. The structural and mechanistic details of cysteine modification have been studied in model enzymes, providing insights into potential transnitrosylation mechanisms.
Resolution and reversibility
In simple terms: The modification can be removed or transferred further, making it a dynamic process.
S-nitrosylation is reversible; the NO group can be removed by reducing agents or transferred to other cysteines, allowing dynamic regulation. This reversibility is crucial for signaling. Enzymes involved in redox homeostasis, such as NADPH oxidase, can influence the cellular environment that governs these reactions.
Key Genes Involved in GO:0035606 peptidyl-cysteine S-trans-nitrosylation
The following genes and proteins are involved in or related to cysteine oxidation and redox processes that inform our understanding of peptidyl-cysteine S-trans-nitrosylation.
| Gene | Major Role | Research Relevance |
|---|---|---|
| SUMF1 | Formylglycine-generating enzyme (FGE) catalyzes cysteine oxidation in sulfatases | Model for copper-dependent cysteine oxidation chemistry |
| TglHI | Peptidyl-(S)-2-mercaptoglycine synthase involved in 3-thiaglutamate biosynthesis | Substrate recognition and cysteine modification |
| AlbA | Thioether bond formation in sactipeptide biosynthesis | Structural insights into cysteine-based modifications |
| EpiD | Peptidyl-cysteine decarboxylase in epidermin biosynthesis | Cysteine decarboxylation model |
| NOX1 | NADPH oxidase component | Redox homeostasis and reactive oxygen species production |
| NOX2 | NADPH oxidase component | Redox signaling in immune cells |
| NOX4 | NADPH oxidase component | Redox regulation in various tissues |
| NCF1 | Neutrophil cytosolic factor 1, NADPH oxidase subunit | Redox homeostasis |
| NCF2 | Neutrophil cytosolic factor 2, NADPH oxidase subunit | Redox homeostasis |
| NCF4 | Neutrophil cytosolic factor 4, NADPH oxidase subunit | Redox homeostasis |
| CYBA | Cytochrome b-245 alpha chain, NADPH oxidase subunit | Redox homeostasis |
| CYBB | Cytochrome b-245 beta chain, NADPH oxidase subunit | Redox homeostasis |
| GAPDH | Glycolytic enzyme with reactive cysteine | Potential target of S-nitrosylation |
| CASP3 | Caspase-3, apoptotic protease with cysteine active site | Cysteine modification can affect activity |
| PTEN | Phosphatase with active-site cysteine | Redox regulation of phosphatase activity |
| SOD1 | Superoxide dismutase 1 | Redox balance and cysteine modifications |
| TXN | Thioredoxin, redox regulator | Redox homeostasis and cysteine reduction |
How Is peptidyl-cysteine S-trans-nitrosylation Regulated?
The process of peptidyl-cysteine S-trans-nitrosylation is regulated by the cellular redox environment, including the availability of nitric oxide donors and the activity of redox enzymes such as NADPH oxidase. The reversibility of S-nitrosylation suggests that reducing systems, like thioredoxin, can remove the modification, thereby controlling signaling. Additionally, the expression and activity of enzymes that generate or transfer NO groups can influence the extent of transnitrosylation. In fungal pathogens, NADPH oxidase is crucial for maintaining redox homeostasis, indicating that similar regulatory mechanisms may exist in other organisms.
peptidyl-cysteine S-trans-nitrosylation and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| SUMF1 | Sulfatase deficiency disorders (e.g., multiple sulfatase deficiency) | Knockout or point mutation in cell models |
| NOX1 | Inflammatory and metabolic diseases | Overexpression or knockout in epithelial cells |
| NOX2 | Chronic granulomatous disease | Knockout in phagocytes |
| NOX4 | Fibrosis and metabolic disorders | Knock-in of tagged version for localization |
| TXN | Redox-related diseases | Point mutation of active-site cysteines |
Metabolic and inflammatory diseases
Proteomic analysis of saliva from patients with type 2 diabetes mellitus and periodontitis revealed alterations in proteins that may be subject to redox modifications, suggesting a link between cysteine-based modifications and these conditions. Although direct evidence for peptidyl-cysteine S-trans-nitrosylation in these diseases is limited, the broader role of cysteine oxidation in metabolic and inflammatory pathways is supported by these findings.
Infectious diseases
In the fungal pathogen Botrytis cinerea, NADPH oxidase is essential for cellular redox homeostasis and virulence, highlighting the importance of redox regulation in host-pathogen interactions. Cysteine modifications, including potential transnitrosylation, may contribute to these processes, although direct evidence is not yet available.
Cancer and neurodegeneration
While specific studies linking GO:0035606 to cancer or neurodegeneration are not present in the verified citations, the general role of cysteine oxidation and NO signaling in these diseases is well recognized. The mechanistic insights from copper-dependent cysteine oxidation enzymes may inform future research in these areas.
From peptidyl-cysteine S-trans-nitrosylation-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does gene X regulate peptidyl-cysteine S-trans-nitrosylation? | Knockout cell line |
| Does a specific cysteine residue serve as an acceptor in transnitrosylation? | Point mutation (Cys-to-Ser) knock-in |
| How does tagging affect protein localization during transnitrosylation? | Tagged knock-in (e.g., GFP or HA) |
| Does overexpression of a candidate gene enhance transnitrosylation? | Overexpression cell line |
| Which genes are essential for redox homeostasis linked to transnitrosylation? | CRISPR library screening |
| What are the global proteomic changes upon modulation of transnitrosylation? | Bioinformatics and proteomics |
How to Study the peptidyl-cysteine S-trans-nitrosylation Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Mass spectrometry | Identification of S-nitrosylated peptides | Global profiling of transnitrosylation targets |
| Biotin switch assay | S-nitrosylation levels | Detection in cell lysates |
| X-ray crystallography | Three-dimensional structure of enzymes | Mechanistic insights into cysteine oxidation |
| Site-directed mutagenesis | Role of specific cysteine residues | Functional analysis of transnitrosylation |
| CRISPR knockout | Gene function in redox pathways | Loss-of-function studies |
| Proteomics | Protein expression and modification changes | Disease biomarker discovery |
| Enzymatic assays | Catalytic activity of redox enzymes | Characterization of NO transfer |
| Fluorescence microscopy | Subcellular localization of proteins | Tracking transnitrosylation events |
Proteomic detection of S-nitrosylated proteins
Proteomic approaches, such as mass spectrometry-based methods, can identify proteins undergoing S-nitrosylation and potentially transnitrosylation. These methods often involve biotin switch assays or enrichment of nitrosylated peptides. Clinical proteomics has been applied to saliva samples to detect redox-related changes in disease.
Structural and mechanistic studies
X-ray crystallography and biochemical assays have elucidated the mechanisms of cysteine oxidation by enzymes like formylglycine-generating enzyme, providing a template for understanding transnitrosylation chemistry. These studies reveal how copper and oxygen activate cysteine residues for modification.
Genetic manipulation in model organisms
Knockout and knockdown studies in fungi and other organisms have demonstrated the importance of NADPH oxidase in redox homeostasis, which may intersect with transnitrosylation pathways. Such genetic tools allow causal testing of gene function.
Biochemical assays for transnitrosylation
In vitro assays using purified proteins can directly measure NO transfer between cysteines. These assays often employ NO donors and detect changes in S-nitrosylation using fluorescent or radioactive probes.
How CRISPR Can Be Used to Study GO:0035606 peptidyl-cysteine S-trans-nitrosylation
Knockout
CRISPR knockout of genes encoding redox enzymes or candidate transnitrosylation regulators can reveal their necessity in the process. For example, knocking out NADPH oxidase components in fungal pathogens affects redox homeostasis. In human cells, knockout of SUMF1 would disrupt cysteine oxidation chemistry, providing a model to study related pathways.
Point Mutation
Introducing point mutations at specific cysteine codons (e.g., Cys-to-Ser) using CRISPR base editing or homology-directed repair allows testing of individual cysteine residues as donors or acceptors in transnitrosylation. This approach has been used to study cysteine oxidation in enzymes like FGE.
Knock-in
Knock-in of tagged versions of proteins (e.g., GFP, HA) enables visualization and immunoprecipitation of transnitrosylation targets. Tagged knock-in models can also be used to track real-time NO transfer in live cells.
Overexpression
Overexpression of candidate genes via CRISPR activation or cDNA delivery can enhance transnitrosylation activity, allowing gain-of-function studies. This is useful for identifying downstream effects of increased NO transfer.
How EDITGENE Supports peptidyl-cysteine S-trans-nitrosylation Research
Researchers studying peptidyl-cysteine S-trans-nitrosylation-related genes often need to determine whether a candidate gene is causally involved in the process, and CRISPR-based models provide a robust way to test this. EDITGENE offers a suite of services to generate precisely engineered cell lines for such investigations.
Contact EDITGENE today to design your custom CRISPR model for peptidyl-cysteine S-trans-nitrosylation research.
Frequently Asked Questions About peptidyl-cysteine S-trans-nitrosylation
What is peptidyl-cysteine S-trans-nitrosylation?
It is the transfer of a nitric oxide (NO) group from one cysteine residue to another, as defined by GO:0035606.
What genes are involved in peptidyl-cysteine S-trans-nitrosylation?
Genes involved in cysteine oxidation and redox regulation, such as SUMF1 and NADPH oxidase components, are relevant to this process.
How is peptidyl-cysteine S-trans-nitrosylation studied?
It can be studied using proteomic methods, biochemical assays, and CRISPR-based genetic models.
What is the role of cysteine in S-trans-nitrosylation?
Cysteine serves as both the donor and acceptor of the NO group, undergoing reversible modification.
Is peptidyl-cysteine S-trans-nitrosylation reversible?
Yes, S-nitrosylation is reversible, allowing dynamic regulation of protein function.
What diseases are associated with peptidyl-cysteine S-trans-nitrosylation?
It may be linked to metabolic and inflammatory diseases such as diabetes and periodontitis, though direct evidence is still emerging.
How does NADPH oxidase relate to transnitrosylation?
NADPH oxidase is crucial for cellular redox homeostasis, which can influence cysteine modifications including transnitrosylation.
Can CRISPR be used to study peptidyl-cysteine S-trans-nitrosylation?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models enable functional studies of genes in this pathway.
What is the GO ID for peptidyl-cysteine S-trans-nitrosylation?
The GO ID is GO:0035606.
What are synonyms for peptidyl-cysteine S-trans-nitrosylation?
Synonyms include cysteine to cysteine nitrosylation, cysteine-to-cysteine transnitrosylation, protein-to-protein transnitrosylation, and S-transnitrosylation.
Conclusion
Peptidyl-cysteine S-trans-nitrosylation (GO:0035606) is a specific post-translational modification involving the transfer of a nitric oxide group between cysteine residues. While direct studies on this exact process are limited, related research on cysteine oxidation mechanisms and redox regulation provides a solid foundation for understanding its biological significance. The process is likely important in redox signaling and may contribute to diseases such as diabetes and periodontitis. CRISPR-based models offer powerful tools to dissect the genes and pathways involved, and EDITGENE provides comprehensive services to support such research.
References
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- 7. Miarzlou DA et al.. 2019. Structure of formylglycine-generating enzyme in complex with copper and a substrate reveals an acidic pocket for binding and activation of molecular oxygen.. Chem Sci 10(29):7049-7058 PMID: 31588272
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