GO:0120515 fatty acid-CoA ligase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0120515 fatty acid-CoA ligase activity catalyzes the ATP-dependent ligation of a fatty acid with coenzyme A to form fatty acyl-CoA, AMP, and diphosphate.
• Long-chain acyl-CoA synthetases (ACSL1, ACSL3, ACSL4, ACSL5, ACSL6) are the principal enzymes carrying this activity and are central to lipid metabolism, membrane remodeling, and ferroptosis [4,6].
• ACSL4 is a key driver of ferroptosis by enriching membranes with polyunsaturated fatty acyl-CoAs, and its activity is regulated by phosphorylation and metabolic signals [1,2,5].
• Loss of ACSL1 can confer ferroptosis resistance in clear cell renal carcinoma, illustrating context-dependent roles of fatty acid-CoA ligase activity in cancer.
• ACSM1 and related acyl-CoA synthetases modulate oxidative stress and cell death, linking this activity to redox biology and disease.
• CRISPR knockout, point mutation, knock-in, and overexpression models are essential to dissect the causal roles of fatty acid-CoA ligase genes in health and disease [4,6].
Description
Fatty acid-CoA ligase activity (GO:0120515) is a fundamental enzymatic activity that activates fatty acids for diverse metabolic fates. It catalyzes the reaction: a fatty acid + ATP + CoA = a fatty acyl-CoA + AMP + diphosphate. This activity is required for fatty acid trafficking into lipid synthesis, beta-oxidation, and membrane phospholipid remodeling [4,6]. Researchers study this term because it sits at the intersection of energy metabolism, ferroptosis, and cancer biology, with acyl-CoA synthetases emerging as therapeutic targets [4,6]. The importance of this activity is underscored by its evolutionary conservation and its involvement in human diseases ranging from metabolic disorders to cancer [4,6]. Understanding the molecular players and regulatory mechanisms of fatty acid-CoA ligase activity is therefore critical for both basic and translational research.
fatty acid-CoA ligase activity At A Glance
| GO ID | GO:0120515 |
|---|---|
| GO term | fatty acid-CoA ligase activity |
| Ontology | molecular_function |
| Synonym | none |
| Definition | Catalysis of the reaction: a fatty acid + ATP + CoA = a fatty acyl-CoA + AMP + diphosphate. |
| Major function | ATP-dependent activation of fatty acids by ligation to coenzyme A, forming fatty acyl-CoA. |
| Related enzymes | Long-chain acyl-CoA synthetases (ACSL1, ACSL3, ACSL4, ACSL5, ACSL6) and other acyl-CoA synthetases [4,6]. |
| Pathways | Fatty acid metabolism, lipid biosynthesis, beta-oxidation, ferroptosis [4,6]. |
| Disease relevance | Cancer, ferroptosis-related pathologies, metabolic disorders [1,2,3,4,5,6,7,8]. |
What Is GO:0120515?
According to the Gene Ontology, GO:0120515 fatty acid-CoA ligase activity is defined as the catalysis of the reaction: a fatty acid + ATP + CoA = a fatty acyl-CoA + AMP + diphosphate. In other words, it is the ATP-dependent activation of a fatty acid by conjugation to coenzyme A, producing a fatty acyl-CoA thioester, AMP, and pyrophosphate. This activity is a molecular function that enables fatty acids to participate in downstream metabolic pathways such as lipid synthesis, oxidation, and signaling.
Why Is fatty acid-CoA ligase activity Important in Cell Biology?
Fatty acid-CoA ligase activity is essential for cellular lipid homeostasis and energy metabolism. By converting fatty acids into fatty acyl-CoAs, it provides the activated substrates for phospholipid synthesis, triacylglycerol storage, and mitochondrial beta-oxidation [4,6]. This activity also determines the cellular sensitivity to ferroptosis, a form of regulated cell death driven by lipid peroxidation, with ACSL4 being a key contributor [1,2,5]. Dysregulation of fatty acid-CoA ligases is implicated in cancer, metabolic diseases, and degenerative conditions, making this activity a promising target for therapeutic intervention [3,4,6,7,8].
• Provides activated fatty acyl-CoAs for membrane phospholipid remodeling and energy production [4,6].
• ACSL4-mediated fatty acid-CoA ligase activity is required for ferroptosis execution [1,2,5].
• ACSL1 loss promotes ferroptosis resistance in clear cell renal carcinoma.
• ACSM1 downregulation enhances oxidative stress and ferroptotic death.
• Tumor-repopulating cells evade ferroptosis via PCK2-dependent phospholipid remodeling involving acyl-CoA synthetases.
• Fatty acid-CoA ligase activity is linked to intervertebral disc degeneration through ACSL4 lactylation.
• Acyl-CoA synthetases are potential therapeutic targets in cancer and metabolic disorders [4,6].
• The activity is regulated by phosphorylation, e.g., PKCβII phosphorylates ACSL4 to amplify lipid peroxidation.
• NF2-YAP signaling modulates ferroptosis sensitivity via regulation of acyl-CoA synthetases.
• CRISPR-based models enable functional dissection of individual acyl-CoA synthetase genes [4,6].
Molecular Mechanism of fatty acid-CoA ligase activity
Substrate binding and activation
In simple terms: The enzyme grabs a fatty acid and uses ATP to make it reactive.
Fatty acid-CoA ligases bind a fatty acid substrate and ATP, forming a fatty acyl-AMP intermediate with release of pyrophosphate. This two-step reaction activates the fatty acid for subsequent thioester bond formation with coenzyme A.
Catalytic mechanism and product formation
In simple terms: The activated fatty acid is linked to coenzyme A, producing fatty acyl-CoA.
In the second step, the fatty acyl-AMP intermediate reacts with CoA to form fatty acyl-CoA and AMP. This thioester product is the central metabolite for lipid synthesis and oxidation [4,6].
Enzyme families and isoforms
In simple terms: Different enzymes handle different fatty acid lengths and cellular locations.
The acyl-CoA synthetase family includes long-chain (ACSL1, ACSL3, ACSL4, ACSL5, ACSL6), very long-chain (ACSVL), and medium-chain (ACSM) enzymes. Each isoform exhibits distinct substrate preferences and tissue distribution, contributing to specialized metabolic functions [4,6].
Regulation by phosphorylation and signaling
In simple terms: Chemical tags and signaling pathways can turn the enzyme on or off.
ACSL4 activity is regulated by phosphorylation; PKCβII phosphorylates ACSL4 to amplify lipid peroxidation during ferroptosis. Additionally, NF2-YAP signaling influences the expression of acyl-CoA synthetases, thereby modulating ferroptosis sensitivity.
Role in ferroptosis and lipid peroxidation
In simple terms: Certain acyl-CoA products make membranes vulnerable to oxidative damage.
ACSL4 preferentially generates polyunsaturated fatty acyl-CoAs that are incorporated into phospholipids, which are prone to peroxidation and drive ferroptosis [1,2,5]. This links fatty acid-CoA ligase activity directly to cell death pathways.
Key Genes Involved in GO:0120515 fatty acid-CoA ligase activity
The following genes encode enzymes with fatty acid-CoA ligase activity or closely related acyl-CoA synthetases, and they are frequently studied in metabolism and disease research.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ACSL1 | Long-chain acyl-CoA synthetase; activates fatty acids for beta-oxidation and lipid synthesis | Loss confers ferroptosis resistance in clear cell renal carcinoma |
| ACSL3 | Long-chain acyl-CoA synthetase; contributes to lipid droplet formation | Implicated in cancer metabolism and ferroptosis [4,6] |
| ACSL4 | Long-chain acyl-CoA synthetase; generates PUFA-CoAs for phospholipid remodeling | Key driver of ferroptosis; regulated by phosphorylation [1,2,5] |
| ACSL5 | Long-chain acyl-CoA synthetase; involved in triacylglycerol synthesis | Associated with metabolic disorders and cancer [4,6] |
| ACSL6 | Long-chain acyl-CoA synthetase; highly expressed in brain | Linked to neurological functions and lipid metabolism [4,6] |
| ACSM1 | Medium-chain acyl-CoA synthetase | Downregulation promotes oxidative stress and ferroptotic death |
| ACSM2A | Medium-chain acyl-CoA synthetase | Expressed in kidney and liver; roles in metabolic pathways |
| ACSM2B | Medium-chain acyl-CoA synthetase | Associated with metabolic traits |
| ACSM3 | Medium-chain acyl-CoA synthetase | Potential tumor suppressor in some cancers |
| ACSM4 | Medium-chain acyl-CoA synthetase | Expressed in olfactory epithelium |
| ACSM5 | Medium-chain acyl-CoA synthetase | Implicated in lipid metabolism |
| SLC27A1 | Very long-chain acyl-CoA synthetase (FATP1) | Fatty acid transport and activation |
| SLC27A2 | Very long-chain acyl-CoA synthetase (FATP2) | Peroxisomal and ER fatty acid activation |
| SLC27A4 | Very long-chain acyl-CoA synthetase (FATP4) | Skin barrier and lipid metabolism |
| SLC27A5 | Very long-chain acyl-CoA synthetase (FATP5) | Bile acid metabolism |
| SLC27A6 | Very long-chain acyl-CoA synthetase (FATP6) | Heart-specific fatty acid uptake |
| THEM4 | Acyl-CoA thioesterase; modulates acyl-CoA pools | Regulates insulin signaling and lipid metabolism |
How Is fatty acid-CoA ligase activity Regulated?
Fatty acid-CoA ligase activity is regulated at multiple levels. Transcriptional control by lipogenic transcription factors such as SREBP-1c and ChREBP influences ACSL expression. Post-translational modifications, including phosphorylation by PKCβII, directly modulate ACSL4 activity and its ability to promote lipid peroxidation. Signaling pathways such as NF2-YAP regulate the expression of acyl-CoA synthetases, affecting ferroptosis sensitivity. Additionally, metabolic signals like lactate can induce ACSL4 expression and lactylation, linking glycolysis to ferroptosis during intervertebral disc degeneration. These layers of regulation ensure that fatty acid activation is tightly coupled to cellular metabolic state and stress responses.
fatty acid-CoA ligase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ACSL4 | Ferroptosis, intervertebral disc degeneration, cancer | ACSL4 knockout or point-mutant cell lines; ferroptosis induction assays [1,2,5] |
| ACSL1 | Clear cell renal carcinoma, ferroptosis resistance | ACSL1 knockout in ccRCC cell lines; lipid peroxidation measurements |
| ACSM1 | Oxidative stress, ferroptotic death | ACSM1 knockdown or knockout; ROS and lipid ROS detection |
| ACSL3 | Cancer metabolism, lipid droplet formation | ACSL3 overexpression or knockout; lipidomics [4,6] |
| ACSL5 | Metabolic disorders, cancer | ACSL5 knockout models; metabolic profiling [4,6] |
Cancer and ferroptosis
Fatty acid-CoA ligase activity is critically involved in ferroptosis, a form of cell death that can be exploited for cancer therapy. ACSL4 is a key determinant of ferroptosis sensitivity by generating polyunsaturated fatty acyl-CoAs that undergo peroxidation [1,2,5]. In clear cell renal carcinoma, loss of ACSL1 fuels ferroptosis resistance, suggesting that ACSL1 expression status may influence therapeutic outcomes. Tumor-repopulating cells evade ferroptosis via PCK2-dependent phospholipid remodeling, highlighting metabolic adaptations that alter acyl-CoA synthetase activity. These findings position fatty acid-CoA ligases as potential targets to modulate ferroptosis in cancer [4,6].
Intervertebral disc degeneration
Glycolysis-derived lactate induces ACSL4 expression and lactylation, which activates ferroptosis during intervertebral disc degeneration. This links fatty acid-CoA ligase activity to degenerative musculoskeletal diseases and suggests that targeting ACSL4 may be a therapeutic strategy.
Oxidative stress and metabolic disorders
ACSM1 downregulation promotes oxidative stress and ferroptotic death, indicating that medium-chain acyl-CoA synthetases also modulate redox balance. Dysregulation of fatty acid-CoA ligases has been associated with metabolic disorders such as obesity and insulin resistance, though the precise mechanisms remain under investigation [4,6].
From fatty acid-CoA ligase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of ACSL4 protect against ferroptosis? | ACSL4 knockout cell lines (e.g., CRISPR-Cas9) [1,2] |
| Does a specific phosphorylation site on ACSL4 regulate its pro-ferroptotic function? | ACSL4 point mutant (phospho-dead or phospho-mimetic) knock-in |
| Can ACSL1 expression status predict ferroptosis sensitivity in ccRCC? | ACSL1 knockout and overexpression in ccRCC cell lines |
| What is the impact of ACSM1 downregulation on oxidative stress? | ACSM1 knockout or knockdown; ROS assays |
| How does PCK2-dependent phospholipid remodeling affect ferroptosis? | PCK2 knockout or overexpression; lipidomics and ferroptosis assays |
| Does NF2-YAP signaling regulate acyl-CoA synthetase expression? | YAP knockout or overexpression; gene expression profiling |
How to Study the fatty acid-CoA ligase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| CRISPR knockout screening | Gene essentiality and ferroptosis sensitivity | Identify novel regulators of fatty acid-CoA ligase activity [4,6] |
| Lipidomics | Fatty acyl-CoA and phospholipid species | Quantify products of fatty acid-CoA ligase activity [1,4] |
| Phosphoproteomics | Phosphorylation sites on ACSL enzymes | Map regulatory modifications |
| Ferroptosis assays | Cell death and lipid peroxidation | Assess functional consequences of ACSL4 activity [1,2,5] |
| RNA-seq | Transcriptional changes in acyl-CoA synthetases | Evaluate expression regulation [4,6] |
| Western blot | Protein expression and modification | Validate knockout or overexpression [3,8] |
| Immunofluorescence | Subcellular localization | Determine organelle-specific functions |
| Co-immunoprecipitation | Protein-protein interactions | Identify regulatory partners [2,5] |
CRISPR-Cas9 knockout screens
Genome-wide CRISPR knockout screens can identify genes whose loss alters ferroptosis sensitivity or lipid metabolism, uncovering novel regulators of fatty acid-CoA ligase activity [4,6].
Lipidomics and metabolomics
Mass spectrometry-based lipidomics quantifies fatty acyl-CoA species and phospholipid composition, providing direct readouts of fatty acid-CoA ligase activity [1,4].
Phosphoproteomics
Phosphoproteomic profiling can reveal phosphorylation events on ACSL enzymes, such as PKCβII-mediated ACSL4 phosphorylation, linking signaling to activity.
Ferroptosis assays
Cell viability assays with ferroptosis inducers (e.g., RSL3, erastin) and lipid peroxidation sensors (e.g., C11-BODIPY) measure the functional impact of fatty acid-CoA ligase activity [1,2,5].
How CRISPR Can Be Used to Study GO:0120515 fatty acid-CoA ligase activity
Knockout
CRISPR-Cas9 knockout of ACSL4 or ACSL1 is used to determine their causal roles in ferroptosis and lipid metabolism. For example, ACSL4 knockout confers resistance to ferroptosis inducers, while ACSL1 loss promotes ferroptosis resistance in ccRCC [1,2,3].
Point Mutation
Point mutations can be introduced to study specific phosphorylation sites or catalytic residues. For instance, mutating the PKCβII phosphorylation site on ACSL4 can reveal its importance in amplifying lipid peroxidation.
Knock-in
Knock-in of tagged or mutant alleles allows precise tracking and functional analysis. A tagged ACSL4 knock-in can be used to study its localization and interactome in live cells.
Overexpression
Overexpression of ACSL4 or other acyl-CoA synthetases can sensitize cells to ferroptosis, providing gain-of-function evidence for their role in lipid peroxidation [1,5].
How EDITGENE Supports fatty acid-CoA ligase activity Research
Researchers studying fatty acid-CoA ligase activity-related genes often need to determine whether a candidate gene is causally involved in metabolic or disease phenotypes. This requires precise genetic models that can isolate the contribution of individual acyl-CoA synthetases.
Contact EDITGENE today to design your custom CRISPR model for fatty acid-CoA ligase activity research.
Frequently Asked Questions About fatty acid-CoA ligase activity
What is fatty acid-CoA ligase activity?
Fatty acid-CoA ligase activity (GO:0120515) is the ATP-dependent catalysis of fatty acid conjugation to coenzyme A, forming fatty acyl-CoA, AMP, and diphosphate.
What genes are involved in fatty acid-CoA ligase activity?
Key genes include ACSL1, ACSL3, ACSL4, ACSL5, ACSL6, and ACSM family members, as well as SLC27A (FATP) genes [4,6].
How is fatty acid-CoA ligase activity related to ferroptosis?
ACSL4 generates polyunsaturated fatty acyl-CoAs that are incorporated into phospholipids and undergo peroxidation, driving ferroptosis [1,2,5].
What diseases are associated with fatty acid-CoA ligase activity?
It is implicated in cancer, intervertebral disc degeneration, metabolic disorders, and oxidative stress-related conditions [1,2,3,4,6,7,8].
How can I study fatty acid-CoA ligase activity in the lab?
Common methods include CRISPR knockout, lipidomics, ferroptosis assays, and phosphoproteomics [1,2,4].
What is the role of ACSL4 in cancer?
ACSL4 promotes ferroptosis and is a key determinant of sensitivity to ferroptosis inducers, making it a potential therapeutic target [1,2,5].
Does ACSL1 loss affect ferroptosis?
Yes, loss of ACSL1 fuels ferroptosis resistance in clear cell renal carcinoma.
How is ACSL4 regulated?
ACSL4 is regulated by phosphorylation (e.g., by PKCβII) and by metabolic signals such as lactate, which induces its expression and lactylation [1,2].
What are the products of fatty acid-CoA ligase activity?
The products are fatty acyl-CoA, AMP, and diphosphate.
Can CRISPR be used to model fatty acid-CoA ligase activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression are powerful approaches to study these genes [4,6].
Conclusion
Fatty acid-CoA ligase activity (GO:0120515) is a central molecular function that activates fatty acids for diverse metabolic and signaling pathways. Its dysregulation is linked to ferroptosis, cancer, and degenerative diseases, with ACSL4 and ACSL1 serving as key examples [1,2,3,4,5,6,7,8]. Understanding the mechanisms and regulation of this activity requires precise genetic models and functional assays. EDITGENE offers comprehensive CRISPR services to support research on fatty acid-CoA ligase activity and its role in health and disease.
References
- 1. Sun K et al.. 2025. Glycolysis-Derived Lactate Induces ACSL4 Expression and Lactylation to Activate Ferroptosis during Intervertebral Disc Degeneration.. Adv Sci (Weinh) 12(21):e2416149 PMID: 40171826
- 2. Zhang HL et al.. 2022. PKCβII phosphorylates ACSL4 to amplify lipid peroxidation to induce ferroptosis.. Nat Cell Biol 24(1):88-98 PMID: 35027735
- 3. Wang S et al.. 2025. Loss of ACSL1 fuels ferroptosis resistance in clear cell renal carcinoma.. Cancer Biol Ther 26(1):2567815 PMID: 41054261
- 4. Deng X et al.. 2025. Long-chain acyl-CoA synthetases: biological functions, diseases and therapeutic targets.. Mol Biomed 6(1):117 PMID: 41288931
- 5. Wu J et al.. 2019. Intercellular interaction dictates cancer cell ferroptosis via NF2-YAP signalling.. Nature 572(7769):402-406 PMID: 31341276
- 6. Quan J et al.. 2021. ACSL family: The regulatory mechanisms and therapeutic implications in cancer.. Eur J Pharmacol 909:174397 PMID: 34332918
- 7. Li Z et al.. 2024. Tumor-repopulating cells evade ferroptosis via PCK2-dependent phospholipid remodeling.. Nat Chem Biol 20(10):1341-1352 PMID: 38720107
- 8. Zhang J et al.. 2024. RIPK4 promotes oxidative stress and ferroptotic death through the downregulation of ACSM1.. Proc Natl Acad Sci U S A 121(40):e2410628121 PMID: 39316049
