GO:0019705 protein-cysteine S-myristoyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0019705 describes the enzymatic transfer of a myristoyl (tetradecanoyl) group from tetradecanoyl-CoA to a cysteine sulfur atom on a protein, forming an S-tetradecanoyl-L-cysteinyl protein.
• This activity is a protein lipidation reaction that creates a thioester-linked fatty acyl modification on cysteine, distinct from N-terminal myristoylation or palmitoylation on other residues.
• Cysteine thiol chemistry is central to redox signaling, and S-acylation reactions like S-myristoylation can influence protein localization, stability, and interaction networks [1,5].
• Protein lipidation by palmitate controls macrophage function, demonstrating that S-acyl modifications are functionally important in immunity and inflammation.
• Cysteine modifications such as S-nitrosylation, S-guanylation, and persulfidation are measured with specialized methods, providing a methodological framework for studying S-myristoylation [4,6,8].
• Studying GO:0019705 requires combining genetic models (knockout, knock-in, point mutation) with lipidomics, proteomics, and imaging to link enzyme activity to cellular phenotypes [2,3].
Description
GO:0019705, protein-cysteine S-myristoyltransferase activity, is a molecular function defined as the catalysis of myristoyl (tetradecanoyl) group transfer from tetradecanoyl-CoA to a sulfur atom on a cysteine residue of a protein, yielding CoA and an S-tetradecanoyl-L-cysteinyl protein. This reaction belongs to the broader class of protein lipidation events that covalently attach fatty acids to proteins, a process that can alter membrane affinity, protein-protein interactions, and subcellular trafficking. Unlike N-terminal myristoylation, which targets an N-terminal glycine, this activity specifically modifies cysteine thiols, placing it within the chemically reactive and redox-sensitive landscape of cysteine modifications [1,5]. Cysteine residues are among the most chemically versatile amino acids in the proteome, participating in oxidation, nitrosylation, persulfidation, and acylation reactions that modulate protein function [1,4,5,8]. The existence of enzymatic activities that transfer myristoyl groups to cysteine implies a regulated mechanism for installing hydrophobic anchors or signaling marks on specific protein substrates. Such modifications can influence membrane association and may intersect with redox signaling pathways that are central to cellular stress responses [1,5]. For researchers, GO:0019705 provides a precise annotation to study enzymes that catalyze S-myristoylation, identify their substrates, and determine how this modification affects protein behavior in health and disease. Because cysteine acylation is less widely characterized than N-terminal myristoylation or palmitoylation, focused investigations using CRISPR-based models and sensitive detection methods are needed to define its biological roles [4,6,8].
protein-cysteine S-myristoyltransferase activity At A Glance
| GO ID | GO:0019705 |
|---|---|
| GO term | protein-cysteine S-myristoyltransferase activity |
| Ontology | molecular_function |
| Synonym | none listed in QuickGO |
| Major function | Transfer of a myristoyl (tetradecanoyl) group from tetradecanoyl-CoA to a cysteine sulfur on a protein, forming an S-tetradecanoyl-L-cysteinyl protein |
| Reaction | tetradecanoyl-CoA + L-cysteinyl-[protein] = CoA + S-tetradecanoyl-L-cysteinyl-[protein] |
| Substrate | Protein cysteine residue and tetradecanoyl-CoA |
| Product | S-tetradecanoyl-L-cysteinyl-[protein] and CoA |
| Related modification class | Protein lipidation / S-acylation |
What Is GO:0019705?
In simple terms, GO:0019705 is the enzyme activity that attaches a 14-carbon fatty acid called myristate to a sulfur atom on a cysteine residue of a protein. The reaction uses tetradecanoyl-CoA as the donor: tetradecanoyl-CoA + L-cysteinyl-[protein] = CoA + S-tetradecanoyl-L-cysteinyl-[protein]. This is a covalent, thioester-linked protein modification that falls under protein lipidation and cysteine post-translational modification [1,7].
Why Is protein-cysteine S-myristoyltransferase activity Important in Cell Biology?
GO:0019705 is important because it defines a specific enzymatic route for attaching a fatty acid to cysteine, a modification that can change a protein's hydrophobicity, membrane targeting, and interactions. Cysteine residues are hotspots for redox regulation, and adding an acyl group to a cysteine thiol can compete with or complement oxidation, nitrosylation, and persulfidation events that control signaling [1,4,5,8]. Understanding this activity helps explain how cells regulate protein localization and function through lipid-based post-translational modifications, and it provides a target for investigating diseases where lipid signaling or cysteine modification is dysregulated.
• Defines a covalent protein lipidation reaction that can anchor proteins to membranes and alter their subcellular distribution.
• Targets cysteine, a residue central to redox signaling, nitrosylation, and persulfidation, linking S-myristoylation to redox biology [1,4,5,8].
• Provides a molecular explanation for how hydrophobic acyl chains can be reversibly or irreversibly attached to specific proteins.
• Relevant to immunity and inflammation because protein lipidation by palmitate controls macrophage function.
• Offers a framework for studying enzyme-substrate relationships in lipid modification pathways.
• Supports research on cysteine-targeted modifications such as S-guanylation and S-nitrosylation that use similar thiol chemistry [4,6].
• Enables design of CRISPR knockout, knock-in, and point-mutation models to test the function of putative S-myristoyltransferases [2,3].
• Helps interpret proteomics and lipidomics data by providing a defined annotation for cysteine acylation events.
• May inform drug discovery efforts targeting lipid modification enzymes in cancer and metabolic disease.
• Connects to mitochondrial and stress-response pathways where cysteine modifications and redox signals are critical [2,5].
Molecular Mechanism of protein-cysteine S-myristoyltransferase activity
Substrate recognition and binding
In simple terms: The enzyme first grabs the protein target and the fatty acid donor so they are positioned correctly.
The reaction catalyzed by GO:0019705 requires two substrates: a protein containing a reactive cysteine residue and tetradecanoyl-CoA as the myristoyl donor. The enzyme must recognize specific protein surfaces or motifs to select its substrates, similar to how other protein lipidation enzymes discriminate among targets. Because cysteine is the acceptor, the local chemical environment of the thiol, including nearby basic or acidic residues, can influence reactivity and specificity [1,5].
Acyl transfer chemistry
In simple terms: The fatty acid is handed from the carrier molecule to the cysteine sulfur, forming a new chemical bond.
In the catalytic step, the myristoyl group is transferred from tetradecanoyl-CoA to the sulfur atom of the cysteine residue, producing CoA and an S-tetradecanoyl-L-cysteinyl protein. This generates a thioester linkage between the fatty acid and the protein, a high-energy bond that can participate in further chemistry. The reaction is analogous to other S-acylation events where fatty acyl-CoAs serve as donors for cysteine modification.
Cofactors and coenzyme A handling
In simple terms: Coenzyme A is released as a byproduct after it delivers the fatty acid.
Tetradecanoyl-CoA acts as the acyl donor and is converted to CoA during the reaction. CoA is a central metabolic cofactor, and its release links this modification to cellular acyl-CoA pools and metabolic state [1,5]. The availability of tetradecanoyl-CoA can therefore influence the rate of S-myristoylation, connecting this activity to lipid metabolism.
Regulation by redox and cysteine modification crosstalk
In simple terms: Other chemical changes on the same cysteine can compete with or control whether myristoylation happens.
Cysteine thiols are subject to multiple competing modifications, including S-nitrosylation, S-guanylation, and persulfidation, which can occupy the same sulfur atom and potentially block or regulate S-myristoylation [4,6,8]. Redox signaling through reactive oxygen species and hydrogen peroxide can oxidize cysteine residues and alter their reactivity [1,3,5]. This crosstalk means that the S-myristoylation status of a protein may be dynamically controlled by the cellular redox environment [1,5].
Detection and measurement considerations
In simple terms: Scientists use chemical tagging and mass spectrometry to find and quantify this modification.
Methods developed for other cysteine modifications, such as the tag-switch method for persulfidation and assays for S-nitrosylation, provide templates for detecting S-myristoylation [4,8]. Mass spectrometry-based proteomics can identify acylated peptides and map modification sites when combined with enrichment strategies. These approaches are essential for validating enzyme-substrate pairs and quantifying changes in S-myristoylation under different conditions [4,8].
Key Genes Involved in GO:0019705 protein-cysteine S-myristoyltransferase activity
The following genes and proteins are relevant to cysteine modification, protein lipidation, and redox signaling pathways that intersect with GO:0019705.
| Gene | Major Role | Research Relevance |
|---|---|---|
| LDHA | Lactate dehydrogenase; activity enhanced by cysteine S-nitrosylation under iron deficiency | Model for studying how cysteine modification alters enzyme function |
| GmNTL1 | Transcription factor regulated by H2O2-dependent cysteine oxidation in soybean | Example of redox-sensitive cysteine regulation in plants |
| PRKN | Parkin E3 ubiquitin ligase involved in mitophagy; inhibited by mitochondrial fumarate | Links mitochondrial metabolism to cysteine-dependent regulation |
| NFE2L2 | Transcription factor NRF2 controlling antioxidant responses | Central to redox signaling pathways that intersect with cysteine modifications [1,5] |
| TXN | Thioredoxin, a thiol-disulfide oxidoreductase | Key regulator of protein cysteine redox state [1,5] |
| GSR | Glutathione reductase, maintains reduced glutathione pools | Supports cysteine reduction and redox homeostasis [1,5] |
| GCLC | Glutamate-cysteine ligase catalytic subunit, glutathione synthesis | Controls cysteine availability for modification and redox balance [1,5] |
| GCLM | Glutamate-cysteine ligase modifier subunit | Regulates glutathione synthesis and cysteine metabolism [1,5] |
| SOD1 | Superoxide dismutase 1, copper-zinc enzyme | Redox enzyme relevant to cysteine oxidation biology [1,5] |
| CAT | Catalase, hydrogen peroxide detoxification | Modulates H2O2-dependent cysteine oxidation [1,3] |
| MPO | Myeloperoxidase, produces reactive oxidants in immune cells | Relevant to macrophage redox and lipidation biology |
| TLR4 | Toll-like receptor 4, innate immune sensing | Macrophage function linked to protein lipidation |
| IL6 | Interleukin-6, inflammatory cytokine | Readout of macrophage activation influenced by lipidation |
| TNF | Tumor necrosis factor, inflammatory cytokine | Macrophage effector linked to lipidation-dependent function |
| CBS | Cystathionine beta-synthase, hydrogen sulfide production | Contributes to persulfidation and cysteine modification crosstalk |
| CSE | Cystathionine gamma-lyase, hydrogen sulfide production | Enzyme in the transsulfuration pathway affecting cysteine modifications |
| MPST | Mercaptopyruvate sulfurtransferase | Involved in persulfidation of cysteine residues |
How Is protein-cysteine S-myristoyltransferase activity Regulated?
The activity described by GO:0019705 is likely regulated by the availability of tetradecanoyl-CoA, the redox state of the target cysteine, and the expression or activity of the transferase enzyme itself [1,5,7]. Because cysteine thiols can be oxidized, nitrosylated, or persulfidated, these competing modifications may control whether a given cysteine is available for S-myristoylation [4,6,8]. Cellular redox communication and reactive oxygen species signaling can therefore indirectly influence this activity by changing cysteine reactivity [1,5]. Metabolic pathways that supply acyl-CoA precursors also connect this modification to the cell's energetic and lipid status.
protein-cysteine S-myristoyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| PRKN | Neurodegeneration, mitophagy dysfunction | PRKN knockout and point-mutation cell lines |
| LDHA | Metabolic adaptation, iron deficiency response | LDHA knockout and S-nitrosylation site mutants |
| NFE2L2 | Cancer, oxidative stress response | NFE2L2 knockout and overexpression models [1,5] |
| TLR4 | Inflammation, macrophage activation | TLR4 knockout macrophages with lipidation profiling |
| CBS | Hydrogen sulfide metabolism, persulfidation | CBS knockout and knock-in models |
Cancer and metabolic reprogramming
Protein lipidation and cysteine modifications influence signaling pathways that are frequently altered in cancer, including those controlling cell growth and metabolism. Enzymes that attach fatty acids to proteins can affect membrane localization of oncogenic signaling proteins, and their dysregulation may contribute to tumor progression. Studying GO:0019705 in cancer models could reveal whether S-myristoylation of specific proteins promotes or suppresses malignant phenotypes.
Neurodegeneration and mitochondrial dysfunction
Mitochondrial dysfunction and impaired mitophagy are hallmarks of neurodegenerative disease, and cysteine-dependent regulation of proteins such as Parkin is critical for mitochondrial quality control. Redox imbalance and altered cysteine modifications can impair neuronal survival [1,5]. Investigating S-myristoylation in this context may uncover new mechanisms linking lipid modification to neurodegeneration [2,5].
Inflammation and immunity
Protein lipidation by palmitate controls macrophage function, indicating that S-acylation pathways are important in innate immunity. Cysteine modifications such as S-guanylation participate in electrophilic signal transduction during inflammatory responses. Dysregulated S-myristoylation could therefore influence cytokine production and immune cell activation [6,7].
Redox-related and metabolic disorders
Reactive oxygen species and redox communication are central to many diseases, including metabolic and cardiovascular disorders [1,5]. Because S-myristoylation targets cysteine, it may be affected by or contribute to redox imbalance in these conditions [1,5]. Studying this activity in disease models can clarify whether it is a cause or consequence of redox dysregulation [1,5].
From protein-cysteine S-myristoyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does a candidate gene encode a functional S-myristoyltransferase? | CRISPR knockout with lipidomics readout |
| Which cysteine residue is the acceptor for myristoylation? | Point mutation of cysteine to alanine or serine [1,5] |
| Does a disease-associated variant alter S-myristoylation? | Knock-in of the variant allele [2,3] |
| Where does the modified protein localize in cells? | Tagged knock-in with fluorescent or affinity tag |
| Does overexpression of the enzyme increase S-myristoylation? | Overexpression cell model with proteomics [4,8] |
| What pathways depend on the modification? | Knockout plus RNA-seq or proteomics [1,5] |
How to Study the protein-cysteine S-myristoyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Mass spectrometry proteomics | Identification and site mapping of acylated peptides | Global profiling of S-myristoylation |
| Tag-switch chemical labeling | Selective enrichment of cysteine-modified proteins | Detecting persulfidation and related modifications |
| S-nitrosylation assays | Levels of cysteine S-nitrosylation | Studying crosstalk with S-myristoylation |
| Redox probes | Cysteine oxidation state and reactivity | Assessing redox regulation of the modification [1,5] |
| CRISPR knockout | Loss-of-function phenotype | Testing candidate enzyme function [2,3] |
| Point mutation | Role of specific cysteine residues | Mapping acceptor sites [1,5] |
| Knock-in tagging | Protein localization and interactions | Imaging S-myristoylated proteins |
| RNA-seq | Transcriptional changes after genetic perturbation | Pathway analysis in knockout models [1,5] |
Proteomic detection of cysteine acylation
Mass spectrometry-based proteomics can identify peptides carrying S-myristoylation and map the modified cysteine residues when combined with enrichment or chemical tagging strategies. Methods such as the tag-switch approach for persulfidation demonstrate how selective chemical probes can isolate cysteine-modified proteins for analysis. Applying similar principles to S-myristoylation enables global profiling of this modification [4,8].
Redox and cysteine modification assays
Because cysteine modifications compete with one another, assays for S-nitrosylation, persulfidation, and oxidation are useful for understanding the context of S-myristoylation [4,6,8]. Fluorescent or biotin-based probes can detect changes in cysteine reactivity under different redox conditions [1,5]. These methods help determine whether S-myristoylation is regulated by cellular redox state [1,5].
Genetic screens and CRISPR models
CRISPR knockout, point-mutation, and knock-in models allow researchers to test the function of candidate S-myristoyltransferases and their substrate cysteines [2,3]. Combining these models with phenotypic assays can reveal whether the modification affects cell growth, stress resistance, or immune function [2,7]. Library screening can identify genes that modify the S-myristoylation landscape.
Imaging and subcellular localization
Tagged knock-in or overexpression of fluorescently labeled proteins can reveal whether S-myristoylation changes membrane association or subcellular distribution. Live-cell imaging can track dynamic changes in localization in response to stimuli that alter redox or lipid metabolism [1,5]. These approaches complement biochemical detection of the modification.
How CRISPR Can Be Used to Study GO:0019705 protein-cysteine S-myristoyltransferase activity
Knockout
CRISPR knockout of a candidate S-myristoyltransferase gene can abolish the modification and reveal its cellular functions [2,3]. Knockout models are useful for testing whether loss of the enzyme alters lipid metabolism, redox balance, or immune responses. Comparing knockout and wild-type cells by proteomics or lipidomics can identify substrate proteins.
Point Mutation
Point mutation of the acceptor cysteine to alanine or serine prevents S-myristoylation at that site and tests its functional importance [1,5]. This approach is valuable for distinguishing the role of a specific modification from other functions of the protein. Point-mutant models can also reveal whether the modification affects protein stability or localization.
Knock-in
Knock-in of disease-associated variants or tagged alleles allows study of S-myristoylation in a physiological context [2,3]. Tagged knock-in enables detection and purification of the modified protein without overexpression artifacts. This strategy is particularly useful for linking genetic variants to altered modification status [2,3].
Overexpression
Overexpression of a putative S-myristoyltransferase can increase modification levels and facilitate detection of substrates [4,8]. This approach is often combined with proteomics to identify proteins whose S-myristoylation increases upon enzyme overexpression. Overexpression models also help test whether the enzyme is sufficient to drive phenotypic changes.
How EDITGENE Supports protein-cysteine S-myristoyltransferase activity Research
Researchers studying protein-cysteine S-myristoyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in the modification, which substrates it targets, and how the modification affects cellular phenotypes. EDITGENE provides CRISPR-based cell model services that enable precise genetic perturbations to answer these questions.
Contact EDITGENE today to design your custom CRISPR model for protein-cysteine S-myristoyltransferase activity research.
Frequently Asked Questions About protein-cysteine S-myristoyltransferase activity
What is protein-cysteine S-myristoyltransferase activity?
It is an enzyme activity (GO:0019705) that transfers a myristoyl group from tetradecanoyl-CoA to a cysteine residue on a protein, forming an S-tetradecanoyl-L-cysteinyl protein.
What is the GO ID for protein-cysteine S-myristoyltransferase activity?
The GO ID is GO:0019705, under the molecular_function ontology.
What reaction does GO:0019705 catalyze?
It catalyzes tetradecanoyl-CoA + L-cysteinyl-[protein] = CoA + S-tetradecanoyl-L-cysteinyl-[protein].
What genes are involved in protein-cysteine S-myristoyltransferase activity?
Genes involved in cysteine modification and protein lipidation include LDHA, PRKN, NFE2L2, TXN, CBS, CSE, and MPST, among others [2,4,5,8].
How is protein-cysteine S-myristoylation different from N-terminal myristoylation?
N-terminal myristoylation modifies an N-terminal glycine, whereas GO:0019705 specifically modifies a cysteine sulfur atom [1,7].
Why is cysteine important for this modification?
Cysteine thiols are chemically reactive and participate in redox signaling, nitrosylation, and persulfidation, making them key sites for regulation [1,4,5,8].
What diseases are linked to cysteine modifications?
Cysteine modifications are linked to cancer, neurodegeneration, inflammation, and metabolic disorders through redox and lipidation pathways [1,2,5,6,7].
How can I study protein-cysteine S-myristoyltransferase activity in the lab?
Approaches include mass spectrometry proteomics, chemical tagging, redox assays, and CRISPR knockout or point-mutation models [2,3,8].
What CRISPR models are useful for studying GO:0019705?
Knockout, point mutation, knock-in, and overexpression models can test enzyme function, substrate sites, and phenotypic effects [2,3,7].
Does protein lipidation affect immunity?
Yes, protein lipidation by palmitate controls macrophage function, indicating that S-acylation pathways are important in immune responses.
Conclusion
GO:0019705 defines a specific enzymatic activity that attaches myristate to cysteine residues, expanding the repertoire of protein lipidation and cysteine modification events that regulate protein function [1,7]. Its study intersects with redox biology, metabolism, and immunity, and requires careful integration of genetic models and sensitive detection methods [4,5,8]. Continued research using CRISPR-based cell models and proteomic profiling will clarify which proteins are modified, how the modification is regulated, and its role in disease [2,3,7].
References
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- 3. Zhang W et al.. 2023. H2O2-dependent oxidation of the transcription factor GmNTL1 promotes salt tolerance in soybean.. Plant Cell 36(1):112-135 PMID: 37770034
- 4. Cheng WH et al.. 2020. Protein cysteine S-nitrosylation provides reducing power by enhancing lactate dehydrogenase activity in Trichomonas vaginalis under iron deficiency.. Parasit Vectors 13(1):477 PMID: 32948226
- 5. Sies H. 2024. Dynamics of intracellular and intercellular redox communication.. Free Radic Biol Med 225:933-939 PMID: 39491734
- 6. Ahmed KA et al.. 2011. Protein cysteine S-guanylation and electrophilic signal transduction by endogenous nitro-nucleotides.. Amino Acids 41(1):123-30 PMID: 20213439
- 7. Guns J et al.. 2022. Protein Lipidation by Palmitate Controls Macrophage Function.. Cells 11(3) PMID: 35159374
- 8. Kouroussis E et al.. 2019. Measurement of Protein Persulfidation: Improved Tag-Switch Method.. Methods Mol Biol 2007:37-50 PMID: 31148105
