GO:0140186 protein N-acyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0140186 protein N-acyltransferase activity describes the catalysis of an acyl-CoA and a lysyl residue on a protein to form an N6-acyl-L-lysyl protein plus CoA and H+.
• The reaction is central to diverse biological processes including sphingolipid metabolism, bile acid conjugation, and protein acylation.
• Key enzymes include ceramide synthases (e.g., CerS6), bile salt hydrolase, and N-acetylglutamate synthase, which use acyl-CoA substrates to modify proteins or small molecules.
• Dysregulation of protein N-acyltransferase activity is linked to inflammation, metabolic disorders, and kidney disease.
• CRISPR-based knockout, point mutation, and knock-in models are essential for dissecting the causal roles of these enzymes.
• EDITGENE provides comprehensive CRISPR services to study protein N-acyltransferase activity in disease models.
Description
Protein N-acyltransferase activity (GO:0140186) is a molecular function that catalyzes the transfer of an acyl group from an acyl-CoA donor to the epsilon-amino group of a lysine residue on a protein, yielding an N6-acyl-L-lysyl protein, CoA, and a proton. This modification, known as protein acylation, is a reversible post-translational modification that can alter protein localization, stability, and interactions. The reaction is fundamental to many cellular processes, including lipid metabolism, signal transduction, and immune regulation. Researchers study this activity to understand how cells sense and respond to metabolic cues, and how its dysregulation contributes to diseases such as inflammation, diabetes, and cancer. The enzyme family includes ceramide synthases, which acylate sphingoid bases to form ceramides, and bile salt hydrolase, which conjugates bile acids with amines. These enzymes are attractive targets for therapeutic intervention, and CRISPR-based models are invaluable for functional studies.
protein N-acyltransferase activity At A Glance
| GO ID | GO:0140186 |
|---|---|
| GO term | protein N-acyltransferase activity |
| Ontology | molecular_function |
| Synonym | protein acyltransferase activity |
| Definition | Catalysis of the reaction: an acyl-CoA + L-lysyl-[protein] = N6-acyl-L-lysyl-[protein] + CoA + H+. |
| Major function | Transfer of acyl groups to lysine residues on proteins, modulating their activity and interactions. |
| Related enzymes | Ceramide synthases (e.g., CerS6), bile salt hydrolase, N-acetylglutamate synthase. |
| Associated processes | Sphingolipid metabolism, bile acid conjugation, protein acylation, inflammation. |
What Is GO:0140186?
According to the Gene Ontology, protein N-acyltransferase activity (GO:0140186) is defined as the catalysis of the reaction: an acyl-CoA + L-lysyl-[protein] = N6-acyl-L-lysyl-[protein] + CoA + H+. In simpler terms, it is an enzymatic activity that attaches an acyl group to a lysine residue on a protein, using acyl-CoA as the donor. This modification can regulate protein function and is involved in various metabolic and signaling pathways.
Why Is protein N-acyltransferase activity Important in Cell Biology?
Protein N-acyltransferase activity is crucial for maintaining cellular homeostasis and responding to metabolic stress. It regulates the synthesis of bioactive lipids such as ceramides and bile acids, which are key signaling molecules in inflammation and metabolism. Dysregulation of this activity has been implicated in a range of diseases, including inflammatory bowel disease, alcoholic liver disease, and diabetic kidney disease. Understanding the molecular mechanisms and identifying specific enzymes involved can lead to novel therapeutic strategies. Moreover, the development of CRISPR-based tools to manipulate these enzymes enables precise functional studies in relevant disease models.
• Regulates sphingolipid metabolism, influencing cell survival and inflammation.
• Modulates bile acid conjugation, affecting lipid absorption and gut microbiota.
• Plays a role in protein acylation, impacting protein function and localization.
• Linked to inflammatory diseases such as IBD and alcoholic liver disease.
• Involved in metabolic disorders including diabetic kidney disease.
• Potential target for therapeutic intervention in cancer and metabolic diseases.
• Enables light-inducible control of enzymatic activity through engineered fusion proteins.
• Essential for understanding host-microbe interactions via bile acid metabolism.
• Provides a mechanism for post-translational regulation of immune responses.
• Facilitates the study of enzyme kinetics and substrate specificity using CRISPR models.
Molecular Mechanism of protein N-acyltransferase activity
Substrate Recognition and Binding
In simple terms: The enzyme first grabs the acyl-CoA and the target protein.
Protein N-acyltransferases recognize specific acyl-CoA donors and target lysine residues on protein substrates. For example, ceramide synthases (CerS) utilize acyl-CoAs of varying chain lengths to acylate sphingoid bases, forming ceramides. Bile salt hydrolase catalyzes the conjugation of bile acids with amines, demonstrating broad substrate specificity. The binding specificity is determined by the enzyme's active site architecture, which accommodates the acyl chain and the protein substrate.
Catalytic Mechanism
In simple terms: The enzyme transfers the acyl group from acyl-CoA to the lysine on the protein.
The catalytic mechanism involves the nucleophilic attack of the lysine epsilon-amino group on the thioester carbonyl of acyl-CoA, forming a tetrahedral intermediate that collapses to release CoA and the N6-acyl-L-lysyl protein. This reaction is often facilitated by a catalytic base that deprotonates the lysine, enhancing its nucleophilicity. The reaction produces CoA and a proton as byproducts.
Cofactors and Regulation
In simple terms: Other molecules can help or hinder the enzyme's activity.
Some N-acyltransferases require cofactors or are regulated by post-translational modifications. For instance, the engineered N-acyltransferase-LOV2 domain fusion protein enables light-inducible allosteric control of enzymatic activity, highlighting the potential for optogenetic regulation. Additionally, the activity can be modulated by the availability of acyl-CoA substrates, which are influenced by cellular metabolic state.
Biological Context and Diversity
In simple terms: These enzymes do many different jobs in the cell.
Protein N-acyltransferases participate in diverse pathways. CerS6 links ceramide metabolism to innate immune responses in diabetic kidney disease. Bile salt hydrolase from gut bacteria catalyses the formation of amine-conjugated bile acids, impacting host physiology. N-acetylglutamate synthase is a mammalian enzyme that produces N-acetylglutamate, an essential activator of carbamoyl phosphate synthetase I in the urea cycle. These examples illustrate the broad biological significance of this activity.
Key Genes Involved in GO:0140186 protein N-acyltransferase activity
The following genes encode enzymes with protein N-acyltransferase activity or are closely related to this function, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| CERS6 | Ceramide synthase 6; synthesizes ceramides from sphingoid bases and acyl-CoAs | Linked to diabetic kidney disease and innate immunity |
| CERS5 | Ceramide synthase 5; involved in very long-chain fatty acid metabolism | Implicated in 1-deoxySphingolipid toxicity |
| CERS2 | Ceramide synthase 2; produces very long-chain ceramides | Studied in sphingolipid metabolism and inflammation |
| BHS | Bile salt hydrolase; conjugates bile acids with amines | Gut microbial enzyme affecting bile acid pool and IBD |
| NAGS | N-acetylglutamate synthase; produces N-acetylglutamate | Urea cycle regulation; mutations cause hyperammonemia |
| ASAH1 | Acid ceramidase; hydrolyzes ceramides | Involved in sphingolipid metabolism and inflammation |
| SPTLC1 | Serine palmitoyltransferase subunit 1; initiates sphingolipid synthesis | Target for sphingolipid-related diseases |
| SPTLC2 | Serine palmitoyltransferase subunit 2 | Associated with neuropathy and lipid metabolism |
| KDSR | 3-ketodihydrosphingosine reductase; involved in sphingolipid synthesis | Mutations cause skin disorders |
| DEGS1 | Dihydroceramide desaturase; introduces double bond in ceramides | Regulates ceramide levels and apoptosis |
| SGMS1 | Sphingomyelin synthase 1; converts ceramides to sphingomyelin | Affects inflammation and lipid signaling |
| SGMS2 | Sphingomyelin synthase 2 | Linked to bone disorders and lipid metabolism |
| ACER1 | Alkaline ceramidase 1; hydrolyzes ceramides | Regulates ceramide levels in skin |
| ACER2 | Alkaline ceramidase 2 | Involved in sphingosine-1-phosphate signaling |
| ACER3 | Alkaline ceramidase 3 | Associated with leukodystrophy |
| UGCG | UDP-glucose ceramide glucosyltransferase; synthesizes glucosylceramides | Target for Gaucher disease |
| GBA | Glucocerebrosidase; hydrolyzes glucosylceramide | Mutations cause Gaucher disease |
How Is protein N-acyltransferase activity Regulated?
Protein N-acyltransferase activity is regulated at multiple levels. Substrate availability, particularly acyl-CoA levels, directly influences enzyme activity. Post-translational modifications and allosteric regulation can modulate enzyme function; for example, an engineered N-acyltransferase-LOV2 fusion protein enables light-inducible allosteric control. Additionally, expression levels of the enzymes are transcriptionally regulated in response to metabolic and inflammatory signals. In the gut, microbial bile salt hydrolase activity is influenced by the presence of bile acids and dietary factors.
protein N-acyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CERS6 | Diabetic kidney disease | Knockout mouse or kidney organoids |
| BHS | Inflammatory bowel disease | Gnotobiotic mice colonized with BHS mutants |
| NAGS | Hyperammonemia | Patient-derived iPSCs with point mutations |
| CERS5 | 1-DeoxySphingolipid toxicity | Knockout cell lines and lipidomics |
| ASAH1 | Farber lipogranulomatosis | Knock-in mouse models |
Inflammatory and Metabolic Diseases
Dysregulated protein N-acyltransferase activity contributes to inflammatory and metabolic diseases. IL-10 constrains sphingolipid metabolism to limit inflammation, and loss of this control leads to excessive ceramide production and exacerbated inflammatory responses. In diabetic kidney disease, CerS6 links ceramide metabolism to innate immune responses, promoting kidney injury. These findings suggest that targeting N-acyltransferases could mitigate inflammation and metabolic damage.
Gastrointestinal Disorders
Gut microbial bile salt hydrolase catalyses the formation of amine-conjugated bile acids, which can impact host physiology. In inflammatory bowel disease, gut microbial metabolism of 5-ASA diminishes its clinical efficacy, partly through bile acid transformations. Altered bile acid profiles are also observed in alcoholic liver disease, where farnesoid X receptor signaling is disrupted. Thus, protein N-acyltransferase activity in the gut microbiome is a key modifier of gastrointestinal health.
Neurological and Rare Diseases
Mutations in enzymes with N-acyltransferase activity can cause rare inherited disorders. For example, N-acetylglutamate synthase deficiency leads to hyperammonemia due to impaired urea cycle activation. Additionally, very long-chain fatty acids drive 1-deoxySphingolipid toxicity, implicating ceramide synthases in neurotoxicity. These examples highlight the importance of these enzymes in neurological and metabolic homeostasis.
From protein N-acyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of CERS6 protect against diabetic kidney disease? | CERS6 knockout mouse |
| How does BHS mutation affect bile acid conjugation? | BHS point-mutant bacterial strains |
| Can light control N-acyltransferase activity? | Knock-in of LOV2 domain fusion |
| What is the role of NAGS in urea cycle? | NAGS knockout hepatocytes |
| Does overexpression of CERS5 increase ceramide levels? | CERS5 overexpression cell lines |
| How does IL-10 regulate sphingolipid metabolism? | IL-10 knockout macrophages |
How to Study the protein N-acyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| LC-MS lipidomics | Ceramide and sphingolipid levels | Quantifying N-acyltransferase products |
| Bile acid profiling | Amine-conjugated bile acids | Assessing BHS activity |
| CRISPR knockout screens | Gene essentiality for enzyme activity | Identifying novel regulators |
| Recombinant enzyme assays | Kinetic parameters | Characterizing substrate specificity |
| Western blot | Protein expression and acylation | Validating knockout/overexpression |
| Immunofluorescence | Subcellular localization | Visualizing enzyme distribution |
| RNA-seq | Transcriptional changes | Assessing pathway regulation |
| Optogenetic control | Light-inducible activity | Spatiotemporal manipulation |
Enzymatic Activity Assays
In vitro assays using recombinant enzymes and acyl-CoA substrates can measure N-acyltransferase activity. For example, ceramide synthase activity can be assessed by monitoring the conversion of sphingoid bases to ceramides using mass spectrometry. Bile salt hydrolase activity can be measured by detecting amine-conjugated bile acids.
CRISPR-Based Genetic Screens
Genome-wide CRISPR knockout screens can identify genes required for N-acyltransferase activity or its downstream effects. For instance, screens in immune cells can uncover regulators of sphingolipid metabolism. These screens are powerful for discovering novel components of the pathway.
Lipidomics and Metabolomics
Mass spectrometry-based lipidomics allows comprehensive profiling of ceramides, sphingomyelins, and bile acids. This approach has been used to show that IL-10 constrains sphingolipid metabolism and that BHS produces amine-conjugated bile acids. Metabolomics can also reveal changes in acyl-CoA pools.
Imaging and Reporter Assays
Fluorescent reporters and imaging can visualize protein acylation in live cells. For example, engineered N-acyltransferase-LOV2 fusion proteins enable light-inducible control, which can be monitored by fluorescence. Such tools facilitate spatiotemporal studies of enzyme activity.
How CRISPR Can Be Used to Study GO:0140186 protein N-acyltransferase activity
Knockout
CRISPR knockout of genes encoding N-acyltransferases (e.g., CERS6, BHS) can abolish enzyme activity, enabling loss-of-function studies. For example, CERS6 knockout mice are used to study diabetic kidney disease. Knockout cell lines are valuable for lipidomics and metabolic assays.
Point Mutation
Introducing point mutations in catalytic residues or regulatory domains can dissect enzyme mechanism. For instance, mutating the catalytic cysteine of BHS can reveal its role in bile acid conjugation. Point mutations in NAGS can model hyperammonemia.
Knock-in
Knock-in of tagged or fluorescently labeled enzymes allows tracking of protein localization and interactions. For example, knock-in of a LOV2 domain into an N-acyltransferase creates a light-inducible enzyme. Knock-in of disease-associated mutations can model human disorders.
Overexpression
Overexpression of N-acyltransferases (e.g., CERS5) can increase flux through the pathway, leading to elevated ceramide levels. This approach is useful for gain-of-function studies and for producing sufficient material for biochemical assays.
How EDITGENE Supports protein N-acyltransferase activity Research
Researchers studying protein N-acyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in a specific disease or metabolic pathway. EDITGENE provides a suite of CRISPR-based services to enable precise genetic manipulation in relevant cell models and animal models, accelerating functional validation and therapeutic target discovery.
Contact EDITGENE today to design your custom CRISPR model for protein N-acyltransferase activity research.
Frequently Asked Questions About protein N-acyltransferase activity
What is protein N-acyltransferase activity?
It is an enzymatic activity that transfers an acyl group from acyl-CoA to a lysine residue on a protein, as defined by GO:0140186.
What genes are involved in protein N-acyltransferase activity?
Key genes include CERS6, BHS, NAGS, and other ceramide synthases and bile acid-conjugating enzymes.
How is protein N-acyltransferase activity regulated?
It is regulated by substrate availability, post-translational modifications, and allosteric control, as shown by engineered light-inducible systems.
What diseases are associated with protein N-acyltransferase activity?
Diseases include inflammatory bowel disease, diabetic kidney disease, hyperammonemia, and alcoholic liver disease.
What methods are used to study protein N-acyltransferase activity?
Methods include LC-MS lipidomics, CRISPR screens, enzymatic assays, and imaging.
Can CRISPR be used to study protein N-acyltransferase activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are widely used to dissect gene function.
What is the role of CERS6 in disease?
CERS6 links ceramide metabolism to innate immune responses in diabetic kidney disease.
How does bile salt hydrolase relate to protein N-acyltransferase activity?
Bile salt hydrolase catalyses the formation of amine-conjugated bile acids, a reaction analogous to N-acylation.
What is the function of N-acetylglutamate synthase?
NAGS produces N-acetylglutamate, an essential activator of the urea cycle, and mutations cause hyperammonemia.
How can I create a knockout model for a protein N-acyltransferase gene?
EDITGENE provides custom CRISPR knockout services for genes like CERS6 and BHS, tailored to your experimental needs.
Conclusion
Protein N-acyltransferase activity (GO:0140186) is a fundamental enzymatic function with broad implications for metabolism, immunity, and disease. The diversity of enzymes and substrates underscores its biological importance, and ongoing research continues to uncover new roles. CRISPR-based models are indispensable for functional studies, and EDITGENE offers comprehensive services to support this research.
References
- 1. York AG et al.. 2024. IL-10 constrains sphingolipid metabolism to limit inflammation.. Nature 627(8004):628-635 PMID: 38383790
- 2. Rimal B et al.. 2024. Bile salt hydrolase catalyses formation of amine-conjugated bile acids.. Nature 626(8000):859-863 PMID: 38326609
- 3. Manley S et al.. 2015. Role of farnesoid X receptor and bile acids in alcoholic liver disease.. Acta Pharm Sin B 5(2):158-67 PMID: 26579442
- 4. Mehta RS et al.. 2023. Gut microbial metabolism of 5-ASA diminishes its clinical efficacy in inflammatory bowel disease.. Nat Med 29(3):700-709 PMID: 36823301
- 5. Reynolds JA et al.. 2023. An engineered N-acyltransferase-LOV2 domain fusion protein enables light-inducible allosteric control of enzymatic activity.. J Biol Chem 299(4):103069 PMID: 36841477
- 6. Zhu Z et al.. 2025. CerS6 links ceramide metabolism to innate immune responses in diabetic kidney disease.. Nat Commun 16(1):1528 PMID: 39934147
- 7. Majcher A et al.. 2025. Very long-chain fatty acids drive 1-deoxySphingolipid toxicity.. Nat Commun 16(1):11650 PMID: 41298489
- 8. Morizono H et al.. 2004. Mammalian N-acetylglutamate synthase.. Mol Genet Metab 81 Suppl 1(Suppl 1):S4-11 PMID: 15050968
