GO:0019107 myristoyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0019107 (myristoyltransferase activity) is a molecular function defined as the catalysis of transfer of a myristoyl group (CH3-[CH2]12-CO-) to an acceptor molecule.
• The best-characterized enzyme carrying this activity is N-myristoyltransferase (NMT), which covalently attaches myristate to the N-terminal glycine of target proteins.
• NMT activity is elevated in several cancers, including colorectal tumors, and its inhibition can promote degradation of oncoproteins such as the androgen receptor in prostate cancer.
• NMT is essential in protozoan parasites and is a validated drug target in Leishmania, with thienopyrimidine inhibitors showing on-target activity in intracellular amastigotes.
• Beyond glycine myristoylation, NMT can also catalyze lysine myristoylation, expanding its regulatory repertoire in cancer, immunity, and infection.
• Activity and inhibition of NMT can be profiled proteome-wide using sortase A-based chemical proteomics, enabling target engagement studies.
Description
Myristoyltransferase activity (GO:0019107) is a molecular function that catalyzes the transfer of a myristoyl group (a 14-carbon saturated fatty acid, CH3-[CH2]12-CO-) from myristoyl-CoA to an acceptor molecule, typically the N-terminal glycine of a protein. This co-translational or post-translational modification, known as N-myristoylation, increases protein hydrophobicity and promotes membrane association, which is critical for signaling, trafficking, and protein stability. The enzyme responsible, N-myristoyltransferase (NMT), is conserved from fungi to humans and is essential for viability in many organisms. Because myristoylation regulates key oncogenic and immune pathways, NMT has emerged as a promising therapeutic target in cancer and infectious diseases. Understanding GO:0019107 therefore provides a mechanistic entry point for studying protein lipidation, membrane targeting, and drug discovery.
myristoyltransferase activity At A Glance
| GO ID | GO:0019107 |
|---|---|
| GO term | myristoyltransferase activity |
| Ontology | molecular_function |
| Synonym | (none) |
| Major function | Transfer of a myristoyl (CH3-[CH2]12-CO-) group to an acceptor molecule, typically N-terminal glycine of proteins |
| Enzyme class | Acyltransferase (EC 2.3.1.-) |
| Representative enzyme | N-myristoyltransferase (NMT1, NMT2 in humans) |
| Cofactor/substrate | Myristoyl-CoA as acyl donor |
| Subcellular context | Cytosol and membrane-associated fractions |
What Is GO:0019107?
In our own words, GO:0019107 describes the enzymatic activity that moves a myristoyl group from a donor molecule (usually myristoyl-CoA) onto an acceptor substrate. The acceptor is most commonly the alpha-amino group of an N-terminal glycine residue on a protein, forming an amide bond. This activity is the defining catalytic function of N-myristoyltransferases (NMTs) and is distinct from other acyltransferases that use different fatty acyl chains.
Why Is myristoyltransferase activity Important in Cell Biology?
Myristoyltransferase activity is important because it controls a reversible lipid modification that dictates protein localization, stability, and interaction networks. Dysregulated NMT activity is observed in human cancers, including colorectal tumors, and inhibition of NMT can destabilize oncoproteins such as the androgen receptor in prostate cancer. In infectious disease, NMT is essential for the survival of protozoan parasites like Leishmania, making it a validated drug target. Furthermore, NMT can modify lysine residues, broadening its impact on immunity and cancer biology. Thus, GO:0019107 sits at the crossroads of cell signaling, oncology, and anti-infective therapy.
• Regulates membrane targeting of signaling proteins such as Src-family kinases and G proteins.
• Elevated NMT activity is found in rat and human colonic tumors, linking it to colorectal cancer.
• Inhibition of NMT promotes androgen receptor degradation, offering a therapeutic strategy in prostate cancer.
• NMT is essential in Leishmania and other parasites, and inhibitors show on-target activity in intracellular amastigotes.
• NMT can act as both a glycine and lysine myristoyltransferase, expanding its regulatory roles in cancer and immunity.
• Reduced membrane-associated NMT activity is observed in obese Zucker rat liver, connecting it to metabolic regulation.
• Chemical proteomics tools enable whole-proteome profiling of NMT activity and inhibition.
• NMT is a conserved enzyme with structural and mechanistic insights available from multiple organisms.
• Myristoylation is a co-translational modification that can be studied with metabolic labeling and click chemistry.
• Targeting myristoyltransferase activity is a promising approach for drug discovery in oncology and infectious disease.
What Happens During myristoyltransferase activity?
Substrate recognition and binding
In simple terms: The enzyme grabs its two building blocks: a fatty acid carrier and a target protein.
N-myristoyltransferase (NMT) binds myristoyl-CoA and a protein substrate that typically exposes an N-terminal glycine after removal of the initiator methionine. The enzyme recognizes a consensus sequence (e.g., MGXXXS/T) in the acceptor protein, positioning the alpha-amino group for nucleophilic attack.
Catalytic transfer of myristate
In simple terms: The fatty acid is handed over to the protein, forming a strong chemical bond.
The catalytic mechanism involves nucleophilic attack of the protein's N-terminal glycine alpha-amino group on the thioester carbonyl of myristoyl-CoA, forming a stable amide bond and releasing coenzyme A. This reaction is catalyzed by a conserved catalytic base and occurs in a sequential ordered mechanism.
Membrane association and trafficking
In simple terms: The added fat acts like a sticky anchor that pulls the protein to cell membranes.
Following myristoylation, the hydrophobic myristate moiety inserts into lipid bilayers, promoting membrane association of the modified protein. This facilitates protein-protein interactions and signaling at membranes, as seen for Src-family kinases and ADP-ribosylation factors.
Lysine myristoylation as an alternative
In simple terms: The enzyme can also attach myristate to a different amino acid, lysine, in some proteins.
Recent studies have shown that NMT can catalyze myristoylation of lysine residues in addition to N-terminal glycine, expanding the repertoire of substrates. This lysine myristoylation has been implicated in cancer, immunity, and host-pathogen interactions.
Key Genes Involved in GO:0019107 myristoyltransferase activity
The following genes and proteins are central to myristoyltransferase activity, either as catalytic enzymes, substrates, or regulatory components.
| Gene | Major Role | Research Relevance |
|---|---|---|
| NMT1 | Catalyzes N-myristoylation of proteins; main human NMT isoform | Target in cancer and infectious disease; knockout affects viability |
| NMT2 | Second human NMT isoform with overlapping substrate specificity | Potential redundancy with NMT1; studied in cancer cell lines |
| SRC | N-myristoylated tyrosine kinase; membrane-associated signaling | Myristoylation required for Src function; model for studying NMT substrates |
| YES1 | N-myristoylated Src-family kinase | Implicated in cancer; myristoylation affects localization |
| FYN | N-myristoylated kinase involved in T-cell signaling | Used to study myristoylation-dependent membrane targeting |
| LYN | N-myristoylated kinase in immune cells | Model substrate for NMT activity assays |
| GNAI1 | N-myristoylated G protein alpha subunit | Myristoylation required for membrane anchoring and signaling |
| ARF1 | N-myristoylated small GTPase | Regulates vesicular trafficking; myristoylation essential for function |
| ARF6 | N-myristoylated small GTPase | Controls endocytic recycling; myristoylation studied in cell models |
| Hck | N-myristoylated Src-family kinase in myeloid cells | Myristoylation influences localization and activity |
| BID | N-myristoylated BH3-only protein | Myristoylation triggers mitochondrial membrane targeting in apoptosis |
| GAP43 | N-myristoylated neuronal protein | Myristoylation regulates neurite outgrowth and plasticity |
| MARCKS | N-myristoylated actin-binding protein | Myristoylation affects membrane-cytoskeleton interactions |
| NOS3 | Endothelial nitric oxide synthase; N-myristoylated | Myristoylation targets eNOS to caveolae; studied in cardiovascular models |
| Lyn | N-myristoylated kinase | Model for studying NMT inhibitor effects |
| NMT (Leishmania) | Essential NMT in Leishmania major | Drug target; inhibitors show on-target activity |
| NMT (fungal) | Essential NMT in fungi | Antifungal target; conserved mechanism |
| AR | Androgen receptor; its stability is affected by NMT inhibition | NMT inhibition promotes AR degradation in prostate cancer |
How Is myristoyltransferase activity Regulated?
Myristoyltransferase activity is regulated at multiple levels. Expression of NMT1 and NMT2 can be modulated by growth factors and oncogenic signaling, and NMT activity is elevated in colorectal tumors. In obese Zucker rat liver, membrane-associated NMT activity is reduced, suggesting metabolic regulation. Additionally, NMT activity can be inhibited by small molecules, leading to degradation of substrates like the androgen receptor in prostate cancer. The enzyme's subcellular localization and access to substrates also influence its activity, as myristoylation occurs co-translationally on ribosomes and post-translationally on membranes.
myristoyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| NMT1 | Colorectal cancer | NMT1 knockout or knockdown in HCT116 cells; xenograft models |
| NMT1 | Prostate cancer | NMT1 inhibition in LNCaP cells; AR degradation assays |
| NMT (Leishmania) | Leishmaniasis | Leishmania major amastigote infection models; inhibitor treatment |
| NMT2 | Cancer (potential) | NMT2 knockout in cancer cell lines; substrate profiling |
| NMT1 | Metabolic disorders | Zucker rat liver models; NMT activity assays |
Cancer
Increased N-myristoyltransferase activity has been observed in rat and human colonic tumors, suggesting a role in colorectal carcinogenesis. In prostate cancer, inhibition of NMT activity promotes androgen receptor degradation, highlighting NMT as a therapeutic target. NMT also myristoylates lysine residues on proteins involved in cancer and immunity, further linking it to oncogenic pathways.
Infectious disease
N-myristoyltransferase is essential for the survival of protozoan parasites such as Leishmania. Thienopyrimidine inhibitors of Leishmania NMT show on-target activity in intracellular amastigotes, validating the enzyme as a drug target. NMT is also a potential target in fungal infections due to its essential role in fungal viability.
Metabolic disorders
Reduced membrane-associated N-myristoyltransferase activity has been reported in obese (fa/fa) Zucker rat liver, indicating a possible connection between myristoylation and metabolic dysregulation. This suggests that NMT activity may be relevant to obesity-related liver pathology.
From myristoyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does NMT1 loss affect cancer cell viability? | NMT1 knockout in cancer cell lines (e.g., HCT116, LNCaP) |
| Does a point mutation in NMT catalytic site abolish activity? | Point-mutation knock-in of catalytic residues in NMT1 |
| Can NMT inhibitors induce AR degradation? | Knock-in of tagged AR in prostate cancer cells; inhibitor treatment |
| What is the subcellular localization of myristoylated proteins? | Tagged knock-in of NMT substrates (e.g., GFP-Src) |
| Can NMT overexpression drive transformation? | Overexpression of NMT1 in immortalized cells |
| What is the proteome-wide impact of NMT inhibition? | Sortase A-based chemical proteomics in cells |
How to Study the myristoyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Radioactive NMT assay | Transfer of [3H]myristoyl-CoA to peptide | In vitro enzyme kinetics and inhibitor screening |
| Fluorescent NMT assay | Myristoylation of fluorescent peptides | High-throughput screening of NMT inhibitors |
| Sortase A proteomics | Proteome-wide myristoylation and inhibition | Target engagement and off-target profiling |
| Click chemistry with alkynyl-myristate | Labeling of myristoylated proteins in cells | Visualization and identification of substrates |
| Western blot for AR | Androgen receptor protein levels | Assessing NMT inhibition effects in prostate cancer |
| CRISPR knockout screens | Gene essentiality and synthetic lethality | Identifying NMT pathway interactions |
| Immunofluorescence | Subcellular localization of myristoylated proteins | Membrane targeting studies |
| Mass spectrometry | Identification of myristoylated peptides | Mapping NMT substrate specificity |
Enzymatic activity assays
Myristoyltransferase activity can be measured using radioactive or fluorescent myristoyl-CoA transfer assays with peptide substrates. These assays quantify the transfer of myristate to acceptor peptides and are used to screen inhibitors.
Chemical proteomics and sortase A labeling
Whole-proteome profiling of NMT activity and inhibition can be achieved using sortase A-based labeling, which tags myristoylated proteins for enrichment and mass spectrometry. This method enables target engagement studies in cells.
Metabolic labeling and click chemistry
Myristoylation can be studied by feeding cells with alkynyl-myristic acid analogs, followed by click chemistry conjugation to fluorescent or affinity tags. This allows visualization and identification of myristoylated proteins.
CRISPR-based genetic screens
CRISPR knockout screens can identify genes that modulate sensitivity to NMT inhibitors or that are synthetic lethal with NMT loss. Such screens help uncover pathways that interact with myristoyltransferase activity.
How CRISPR Can Be Used to Study GO:0019107 myristoyltransferase activity
Knockout
CRISPR knockout of NMT1 or NMT2 can be used to study loss of myristoyltransferase activity, revealing effects on cell viability, protein localization, and signaling. For example, NMT1 knockout in cancer cell lines can reduce proliferation and induce apoptosis.
Point Mutation
Point mutations in the catalytic domain of NMT can be introduced to dissect the enzymatic mechanism. For instance, mutating the catalytic base or substrate-binding residues can abolish myristoyltransferase activity and serve as negative controls.
Knock-in
Knock-in of tagged NMT substrates (e.g., GFP-Src) allows real-time tracking of myristoylation-dependent membrane targeting. Tagged knock-in models can also be used to study the effect of NMT inhibitors on substrate localization.
Overexpression
Overexpression of NMT1 or NMT2 can mimic elevated myristoyltransferase activity observed in cancers. Such models are useful for studying oncogenic transformation and for testing NMT inhibitors in a background of high enzyme levels.
How EDITGENE Supports myristoyltransferase activity Research
Researchers studying myristoyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in a specific phenotype, such as cancer cell proliferation or parasite survival. This requires precise genetic models that can knockout, mutate, tag, or overexpress the gene of interest. EDITGENE provides a comprehensive suite of CRISPR services to accelerate such studies.
Contact EDITGENE today to design your custom CRISPR model for myristoyltransferase activity research.
Frequently Asked Questions About myristoyltransferase activity
What is myristoyltransferase activity?
Myristoyltransferase activity (GO:0019107) is the catalysis of transfer of a myristoyl group (CH3-[CH2]12-CO-) to an acceptor molecule, typically the N-terminal glycine of a protein.
What genes are involved in myristoyltransferase activity?
The main genes are NMT1 and NMT2, which encode N-myristoyltransferases. Substrate genes include SRC, YES1, FYN, LYN, GNAI1, ARF1, and ARF6.
What is the function of N-myristoyltransferase?
N-myristoyltransferase catalyzes the covalent attachment of myristate to proteins, promoting membrane association and regulating signaling, trafficking, and stability.
How is myristoyltransferase activity measured?
It can be measured using radioactive or fluorescent myristoyl-CoA transfer assays, chemical proteomics with sortase A, or metabolic labeling with click chemistry.
Is myristoyltransferase activity involved in cancer?
Yes, increased NMT activity is observed in colorectal tumors, and NMT inhibition promotes androgen receptor degradation in prostate cancer.
What diseases are linked to myristoyltransferase activity?
It is linked to cancers such as colorectal and prostate cancer, infectious diseases like leishmaniasis, and metabolic disorders such as obesity-related liver changes.
Can CRISPR be used to study myristoyltransferase activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models allow precise dissection of NMT function in cells and animal models.
What are NMT inhibitors?
NMT inhibitors are small molecules that block myristoyltransferase activity; examples include thienopyrimidine inhibitors active against Leishmania NMT.
Does NMT only modify glycine residues?
No, recent evidence shows NMT can also catalyze lysine myristoylation, expanding its substrate repertoire in cancer and immunity.
How does myristoylation affect protein localization?
Myristoylation adds a hydrophobic anchor that inserts into membranes, directing proteins to specific cellular compartments such as the plasma membrane.
Conclusion
Myristoyltransferase activity (GO:0019107) is a fundamental enzymatic function that controls protein lipidation and membrane targeting. Its dysregulation is implicated in cancer, infectious disease, and metabolic disorders, making it a high-value target for therapeutic development. Understanding the molecular mechanism, key genes, and regulatory networks of myristoyltransferase activity provides a foundation for innovative research and drug discovery.
References
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- 2. Rajala RV et al.. 2000. N-myristoyltransferase.. Mol Cell Biochem 204(1-2):135-55 PMID: 10718634
- 3. Magnuson BA et al.. 1995. Increased N-myristoyltransferase activity observed in rat and human colonic tumors.. J Natl Cancer Inst 87(21):1630-5 PMID: 7563206
- 4. Boutin JA. 1997. Myristoylation.. Cell Signal 9(1):15-35 PMID: 9067626
- 5. Bell AS et al.. 2020. Novel Thienopyrimidine Inhibitors of Leishmania N-Myristoyltransferase with On-Target Activity in Intracellular Amastigotes.. J Med Chem 63(14):7740-7765 PMID: 32575985
- 6. Kosciuk T et al.. 2020. N-Myristoyltransferase as a Glycine and Lysine Myristoyltransferase in Cancer, Immunity, and Infections.. ACS Chem Biol 15(7):1747-1758 PMID: 32453941
- 7. King MJ et al.. 1993. Membrane-associated N-myristoyltransferase activity is reduced in obese (fa/fa) Zucker rat liver.. Biochem Biophys Res Commun 196(2):665-70 PMID: 8240341
- 8. Goya Grocin A et al.. 2019. Whole Proteome Profiling of N-Myristoyltransferase Activity and Inhibition Using Sortase A.. Mol Cell Proteomics 18(1):115-126 PMID: 30341083
