GO:0052816 long-chain fatty acyl-CoA hydrolase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0052816 describes the hydrolysis of long-chain fatty acyl-CoA thioesters (13-22 carbons) into free fatty acids and CoA, a reaction that controls cellular acyl-CoA pools.
• Long-chain acyl-CoA hydrolases are distinct from short-chain and mitochondrial thioesterases and are highly expressed in brain cytosol and liver peroxisomes/mitochondria.
• The reaction regulates lipid signaling, membrane remodeling, and metabolic gene expression by altering the availability of long-chain acyl-CoAs.
• Peroxisome proliferators and metabolic states strongly induce long-chain acyl-CoA hydrolase activity in rodent liver.
• Dysregulation of acyl-CoA metabolism is linked to neurodegeneration, metabolic disorders, and cancer, making this activity a potential therapeutic target.
• CRISPR knockout, point mutation, knock-in, and overexpression models are essential to dissect the causal roles of specific hydrolase genes.
Description
Long-chain fatty acyl-CoA hydrolase activity (GO:0052816) is a molecular function that catalyzes the hydrolysis of long-chain fatty acyl-CoA thioesters into free fatty acids and coenzyme A. This activity is critical for maintaining the balance of acyl-CoA pools, which serve as central intermediates in fatty acid oxidation, lipid synthesis, and protein acylation. The enzyme is widely distributed across tissues, with particularly high activity in the brain and liver, where it modulates lipid signaling and energy homeostasis. Researchers study this activity to understand how cells regulate fatty acid trafficking and how its dysregulation contributes to metabolic and neurological diseases. The reaction is also important in peroxisomal and mitochondrial fatty acid metabolism, where it influences substrate availability for beta-oxidation.
long-chain fatty acyl-CoA hydrolase activity At A Glance
| GO ID | GO:0052816 |
|---|---|
| GO term | long-chain fatty acyl-CoA hydrolase activity |
| Ontology | molecular_function |
| Synonym | long-chain-acyl-CoA hydrolase activity; long-chain acyl coenzyme A hydrolase activity; long-chain acyl-thioester hydrolase activity; long-chain hydrolase activity |
| Definition | Catalysis of the reaction: a long-chain fatty acyl-CoA + H2O = a long-chain fatty acid + CoA + H+. A long-chain fatty acid has an aliphatic tail containing 13 to 22 carbons. |
| Major function | Hydrolysis of long-chain fatty acyl-CoA thioesters to free fatty acids and CoA |
| Substrate specificity | Long-chain fatty acyl-CoAs with aliphatic tails of 13 to 22 carbons |
| Tissue distribution | High activity in brain cytosol and liver peroxisomes/mitochondria |
| Regulation | Induced by peroxisome proliferators in rodent liver |
What Is GO:0052816?
According to the Gene Ontology, GO:0052816 is defined as the catalysis of the reaction: a long-chain fatty acyl-CoA + H2O = a long-chain fatty acid + CoA + H+. A long-chain fatty acid has an aliphatic tail containing 13 to 22 carbons. This activity is also known as long-chain-acyl-CoA hydrolase activity, long-chain acyl coenzyme A hydrolase activity, long-chain acyl-thioester hydrolase activity, and long-chain hydrolase activity.
Why Is long-chain fatty acyl-CoA hydrolase activity Important in Cell Biology?
Long-chain fatty acyl-CoA hydrolase activity is essential for lipid homeostasis because it directly controls the cellular levels of long-chain acyl-CoAs, which are key metabolic intermediates and signaling molecules. By hydrolyzing these thioesters, the enzyme generates free fatty acids that can be used for energy production, membrane synthesis, or signaling, and it also liberates CoA to support other metabolic reactions. Dysregulation of this activity has been implicated in neurological disorders, metabolic syndrome, and cancer, making it a subject of intense research.
• Regulates cellular long-chain acyl-CoA pools, which are central to fatty acid oxidation and lipid synthesis.
• Provides free fatty acids for membrane remodeling and signaling.
• Recycles coenzyme A, a critical cofactor for numerous metabolic enzymes.
• Highly expressed in brain, suggesting roles in neuronal lipid metabolism and signaling.
• Induced by peroxisome proliferators, linking it to peroxisomal proliferation and lipid-lowering responses.
• Potential involvement in metabolic disorders such as obesity and diabetes.
• May influence cancer cell metabolism by altering fatty acid availability.
• Target for understanding neurodegeneration due to brain-specific isoforms.
• Important for microbial and biotechnological production of fatty acid-derived compounds.
• Serves as a model for studying thioesterase mechanisms and substrate specificity.
What Happens During long-chain fatty acyl-CoA hydrolase activity?
Substrate binding and recognition
In simple terms: The enzyme grabs a long-chain fatty acyl-CoA molecule.
The hydrolase binds long-chain fatty acyl-CoA substrates with aliphatic tails of 13 to 22 carbons through a hydrophobic binding pocket that accommodates the acyl chain. This binding is highly specific, discriminating against short-chain and medium-chain acyl-CoAs.
Catalytic hydrolysis
In simple terms: Water breaks the bond between the fatty acid and CoA.
A water molecule attacks the thioester bond, releasing free coenzyme A and a long-chain fatty acid. This reaction is catalyzed by a serine-histidine-aspartate catalytic triad typical of many hydrolases. The reaction is reversible in vitro but favors hydrolysis under physiological conditions.
Product release and cellular fate
In simple terms: The products are released to be used elsewhere in the cell.
The free fatty acid can be directed to beta-oxidation, membrane lipid synthesis, or signaling pathways, while CoA is recycled for other acyl-CoA synthetases. In brain, the released fatty acids may participate in neuronal signaling and membrane maintenance.
Tissue-specific isoforms and localization
In simple terms: Different versions of the enzyme work in different parts of the cell.
Cytosolic and peroxisomal/mitochondrial isoforms exist, with the brain cytosolic form being purified and characterized. Peroxisomal proliferators induce the activity in liver peroxisomes and mitochondria, suggesting distinct regulatory mechanisms.
Key Genes Involved in GO:0052816 long-chain fatty acyl-CoA hydrolase activity
The following genes and proteins are experimentally linked to long-chain fatty acyl-CoA hydrolase activity or its regulation.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ACOT1 | Cytosolic long-chain acyl-CoA hydrolase | Model for studying acyl-CoA pool regulation |
| ACOT2 | Mitochondrial long-chain acyl-CoA hydrolase | Linked to fatty acid oxidation and energy metabolism |
| ACOT7 | Brain-specific long-chain acyl-CoA hydrolase | Implicated in neuronal lipid signaling |
| ACOT9 | Peroxisomal long-chain acyl-CoA hydrolase | Induced by peroxisome proliferators |
| ACOT11 | Thioesterase highly expressed in adipose tissue | Potential role in obesity and insulin resistance |
| ACOT12 | Cytosolic acetyl-CoA hydrolase | Related to acyl-CoA metabolism |
| ACOT13 | Mitochondrial thioesterase | Involved in lipid homeostasis |
| ACSL1 | Long-chain acyl-CoA synthetase | Generates substrates for hydrolases |
| ACSL3 | Long-chain acyl-CoA synthetase | Regulates acyl-CoA pools |
| ACSL4 | Long-chain acyl-CoA synthetase | Linked to ferroptosis and lipid signaling |
| CPT1A | Carnitine palmitoyltransferase 1A | Controls mitochondrial fatty acid import |
| CPT2 | Carnitine palmitoyltransferase 2 | Defects cause acyl-CoA accumulation |
| PPARA | Peroxisome proliferator-activated receptor alpha | Mediates induction of hydrolase activity |
| PPARGC1A | PGC-1alpha coactivator | Regulates mitochondrial biogenesis and lipid metabolism |
| SREBF1 | Sterol regulatory element-binding transcription factor 1 | Controls lipogenic gene expression |
| MLYCD | Malonyl-CoA decarboxylase | Regulates malonyl-CoA levels |
| DGAT1 | Diacylglycerol O-acyltransferase 1 | Competes for acyl-CoA pools |
How Is long-chain fatty acyl-CoA hydrolase activity Regulated?
Long-chain fatty acyl-CoA hydrolase activity is regulated at multiple levels. Peroxisome proliferators such as clofibrate strongly induce the activity in rat liver peroxisomes and mitochondria, likely through PPARalpha-mediated transcriptional activation. In brain, the cytosolic hydrolase is constitutively expressed but may be modulated by developmental and metabolic cues. Additionally, the activity is influenced by substrate availability and the nutritional state, as acyl-CoA levels fluctuate with fasting and feeding. Post-translational modifications and protein-protein interactions may also regulate enzyme activity, though specific mechanisms remain to be fully elucidated.
long-chain fatty acyl-CoA hydrolase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ACOT7 | Neurodegeneration, brain lipid signaling | Neuron-specific knockout mouse |
| ACOT1 | Metabolic syndrome, fatty liver | Liver-specific overexpression |
| ACOT2 | Mitochondrial dysfunction, insulin resistance | Knockout in skeletal muscle cells |
| ACOT11 | Obesity, insulin resistance | Adipose-specific knockout |
| ACSL4 | Ferroptosis, cancer | Point mutation of catalytic residue |
Neurodegeneration and brain lipid metabolism
The brain-specific long-chain acyl-CoA hydrolase ACOT7 is highly expressed in neurons, where it regulates acyl-CoA pools critical for membrane phospholipid synthesis and signaling. Dysregulation of this activity has been proposed to contribute to neurodegenerative conditions such as Alzheimer's disease and Parkinson's disease, although direct causal evidence is still emerging.
Metabolic disorders and obesity
Altered long-chain acyl-CoA hydrolase activity can affect fatty acid oxidation and lipogenesis, influencing energy balance. In obesity and type 2 diabetes, acyl-CoA metabolism is perturbed, and hydrolases may contribute to ectopic lipid accumulation and insulin resistance.
Cancer metabolism
Cancer cells often reprogram lipid metabolism to support rapid proliferation. Long-chain acyl-CoA hydrolases can modulate the availability of fatty acids for membrane synthesis and signaling, and their expression is altered in some cancers. Targeting these enzymes may offer therapeutic opportunities, but further studies are needed.
Peroxisomal disorders
Peroxisomal long-chain acyl-CoA hydrolase activity is induced by peroxisome proliferators and is part of the peroxisomal beta-oxidation machinery. Defects in peroxisomal fatty acid oxidation lead to diseases such as X-linked adrenoleukodystrophy, where acyl-CoA accumulation is toxic.
From long-chain fatty acyl-CoA hydrolase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of ACOT7 alter brain acyl-CoA levels? | ACOT7 knockout mouse or neuronal cell line |
| How does ACOT1 overexpression affect lipid droplets? | Hepatocyte overexpression with tagged ACOT1 |
| What is the catalytic mechanism of ACOT2? | Point mutation of active-site serine |
| Does ACOT11 regulate insulin sensitivity? | Adipose-specific knock-in of human variant |
| Can ACOT9 be induced by PPARalpha agonists? | Reporter knock-in of ACOT9 promoter |
| Does ACOT7 interact with other lipid enzymes? | Knock-in of FLAG-tagged ACOT7 |
How to Study the long-chain fatty acyl-CoA hydrolase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Spectrophotometric thioesterase assay | Release of CoA from acyl-CoA | Enzyme kinetics and inhibitor testing |
| LC-MS lipidomics | Acyl-CoA and fatty acid levels | Metabolic profiling in cells/tissues |
| CRISPR knockout screen | Gene essentiality and lipid stress response | Discovery of novel regulators |
| Western blot | Protein expression and tagging | Validation of knockout/overexpression |
| Immunofluorescence | Subcellular localization | Organelle targeting studies |
| qRT-PCR | mRNA expression levels | Transcriptional regulation by PPARalpha |
| Co-immunoprecipitation | Protein-protein interactions | Identifying binding partners |
| Seahorse assay | Mitochondrial respiration | Functional impact of hydrolase activity |
Enzymatic activity assays
Long-chain acyl-CoA hydrolase activity is typically measured using spectrophotometric or fluorometric assays that monitor the release of CoA from acyl-CoA substrates. These assays use substrates such as palmitoyl-CoA and are performed with tissue homogenates or purified enzymes.
Lipidomics and acyl-CoA profiling
Mass spectrometry-based lipidomics can quantify long-chain acyl-CoA species and free fatty acids in cells or tissues, providing a readout of hydrolase activity in vivo. This approach is valuable for linking enzyme function to metabolic phenotypes.
CRISPR-based genetic screens
Genome-wide CRISPR knockout screens can identify genes that modulate acyl-CoA levels or sensitivity to lipid stress, revealing novel regulators of hydrolase activity. Such screens are powerful for discovering synthetic lethal interactions in cancer cells.
Imaging and subcellular localization
Fluorescent tagging of hydrolases (e.g., GFP fusion) allows visualization of subcellular localization in live cells, confirming peroxisomal, mitochondrial, or cytosolic distribution. Co-localization with organelle markers validates compartmentalization.
How CRISPR Can Be Used to Study GO:0052816 long-chain fatty acyl-CoA hydrolase activity
Knockout
CRISPR knockout of long-chain acyl-CoA hydrolase genes (e.g., ACOT7, ACOT1) enables researchers to assess their contribution to acyl-CoA homeostasis, lipid signaling, and cellular phenotypes. Knockout cell lines can be used in lipidomics and metabolic assays to reveal compensatory mechanisms.
Point Mutation
Introducing point mutations in the catalytic serine or histidine residues of hydrolases abolishes enzymatic activity, allowing separation of catalytic function from scaffolding roles. Such mutants are valuable for dissecting substrate specificity and mechanism.
Knock-in
Knock-in of tagged versions (e.g., FLAG, GFP) of hydrolases facilitates localization, interaction, and activity studies in a physiological context. Knock-in of disease-associated variants can model human mutations in isogenic cell lines.
Overexpression
Overexpression of wild-type or mutant hydrolases in cell lines or animal models can drive lipid remodeling and reveal downstream effects on signaling and metabolism. This approach is useful for testing whether increased hydrolase activity protects against lipid-induced toxicity.
How EDITGENE Supports long-chain fatty acyl-CoA hydrolase activity Research
Researchers studying long-chain fatty acyl-CoA hydrolase activity-related genes often need to determine whether a candidate gene is causally involved in lipid metabolism, signaling, or disease. EDITGENE provides comprehensive CRISPR services to generate precisely engineered cell models, enabling functional validation of hydrolase genes and their variants.
Contact EDITGENE today to design your custom CRISPR model for long-chain fatty acyl-CoA hydrolase activity research.
Frequently Asked Questions About long-chain fatty acyl-CoA hydrolase activity
What is long-chain fatty acyl-CoA hydrolase activity?
It is a molecular function (GO:0052816) that catalyzes the hydrolysis of long-chain fatty acyl-CoA thioesters into free fatty acids and coenzyme A.
What genes are involved in long-chain fatty acyl-CoA hydrolase activity?
Genes such as ACOT1, ACOT2, ACOT7, ACOT9, and ACOT11 encode enzymes with this activity.
What is the difference between long-chain and short-chain acyl-CoA hydrolases?
Long-chain hydrolases act on fatty acyl-CoAs with 13-22 carbons, while short-chain hydrolases prefer shorter chains.
Where is long-chain fatty acyl-CoA hydrolase activity found in the cell?
It is found in the cytosol, peroxisomes, and mitochondria, depending on the isoform.
How is long-chain fatty acyl-CoA hydrolase activity regulated?
It can be induced by peroxisome proliferators and is influenced by nutritional state and substrate availability.
What diseases are associated with long-chain fatty acyl-CoA hydrolase dysfunction?
Neurodegeneration, metabolic disorders, and cancer have been linked to altered acyl-CoA metabolism.
How can I measure long-chain fatty acyl-CoA hydrolase activity in the lab?
Common methods include spectrophotometric thioesterase assays and mass spectrometry-based lipidomics.
What are the substrates of long-chain fatty acyl-CoA hydrolase?
Long-chain fatty acyl-CoAs such as palmitoyl-CoA and oleoyl-CoA are typical substrates.
Can CRISPR be used to study long-chain fatty acyl-CoA hydrolase genes?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression are powerful approaches to dissect gene function.
What is the role of long-chain acyl-CoA hydrolase in brain?
Brain-specific isoforms like ACOT7 regulate neuronal acyl-CoA pools and may influence signaling and membrane homeostasis.
Conclusion
Long-chain fatty acyl-CoA hydrolase activity (GO:0052816) is a fundamental enzymatic function that controls the balance of long-chain acyl-CoAs, key metabolites in energy production, lipid synthesis, and signaling. Its tissue-specific isoforms and subcellular localizations enable fine-tuned regulation of lipid metabolism in brain, liver, and other tissues. Dysregulation of this activity is implicated in neurodegeneration, metabolic disorders, and cancer, making it an attractive target for therapeutic intervention. Continued research using CRISPR-based models and advanced lipidomics will further illuminate its mechanistic roles and disease relevance.
References
- 1. Xue Q et al.. 2023. Copper metabolism in cell death and autophagy.. Autophagy 19(8):2175-2195 PMID: 37055935
- 2. Robichaud S et al.. 2021. Identification of novel lipid droplet factors that regulate lipophagy and cholesterol efflux in macrophage foam cells.. Autophagy 17(11):3671-3689 PMID: 33590792
- 3. Yamada J. 2005. Long-chain acyl-CoA hydrolase in the brain.. Amino Acids 28(3):273-8 PMID: 15731883
- 4. Choi YJ et al.. 2013. Microbial production of short-chain alkanes.. Nature 502(7472):571-4 PMID: 24077097
- 5. Black PN et al.. 2000. Long-chain acyl-CoA-dependent regulation of gene expression in bacteria, yeast and mammals.. J Nutr 130(2S Suppl):305S-309S PMID: 10721893
- 6. Bremer J. 1983. Carnitine--metabolism and functions.. Physiol Rev 63(4):1420-80 PMID: 6361812
- 7. Berge RK et al.. 1984. Enhancement of long-chain acyl-CoA hydrolase activity in peroxisomes and mitochondria of rat liver by peroxisomal proliferators.. Eur J Biochem 141(3):637-44 PMID: 6146524
- 8. Yamada J et al.. 1996. Long-chain acyl-CoA hydrolase from rat brain cytosol: purification, characterization, and immunohistochemical localization.. Arch Biochem Biophys 326(1):106-14 PMID: 8579357
