GO:0016289 acyl-CoA hydrolase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0016289 acyl-CoA hydrolase activity catalyzes the hydrolysis of an acyl-CoA thioester into a free carboxylate, CoA, and a proton.
• This activity is carried out by acyl-CoA thioesterases (ACOTs) and related enzymes that regulate cellular acyl-CoA pools.
• ACOT enzymes are found in peroxisomes, mitochondria, and cytosol, and their activity is influenced by peroxisomal proliferators and dietary status.
• Dysregulation of acyl-CoA hydrolase activity contributes to cancer progression, metabolic disorders, and osteoarthritis.
• ACOT8 and ACOT12 are key examples linked to gemcitabine resistance in pancreatic cancer and suppression of YAP-mediated hepatocarcinogenesis, respectively.
• CRISPR knockout, point mutation, knock-in, and overexpression models are essential to dissect the causal roles of acyl-CoA hydrolase genes in disease.
Description
Acyl-CoA hydrolase activity (GO:0016289) is a molecular function that removes the CoA moiety from acyl-CoA molecules, releasing free fatty acids and CoA. This reaction is fundamental to lipid metabolism, as it controls the intracellular levels of acyl-CoA, which are central intermediates in fatty acid oxidation, lipid synthesis, and signaling. The enzymes responsible, known as acyl-CoA thioesterases (ACOTs), are widely distributed across tissues and organelles, including peroxisomes, mitochondria, and the cytosol. Their activity is dynamically regulated by nutritional and hormonal signals, such as peroxisomal proliferators and diabetes. Researchers study acyl-CoA hydrolase activity because it sits at the crossroads of energy homeostasis and disease. For example, ACOT12 limits glycerolipid biosynthesis to suppress YAP-mediated hepatocarcinogenesis, while ACOT8 promotes gemcitabine resistance in pancreatic ductal adenocarcinoma by modulating lipid metabolism and ferroptosis. In the brain, long-chain acyl-CoA hydrolase activity is critical for neuronal lipid metabolism. Moreover, altered acyl-CoA hydrolase activity has been observed in osteoarthritis and atherosclerosis, highlighting its broad pathophysiological relevance. Understanding the precise molecular mechanisms, regulatory networks, and disease associations of acyl-CoA hydrolase activity requires robust experimental models. This article integrates authoritative GO annotations with verified PubMed literature to provide a comprehensive overview of GO:0016289, its key genes, and the CRISPR-based strategies used to study it.
acyl-CoA hydrolase activity At A Glance
| GO ID | GO:0016289 |
|---|---|
| GO term | acyl-CoA hydrolase activity |
| Ontology | molecular_function |
| Synonym | CoA hydrolase activity |
| Definition | Catalysis of the reaction: an acyl-CoA + H2O = a carboxylate + CoA + H+. |
| Major function | Hydrolysis of acyl-CoA thioesters to free fatty acids and CoA |
| Representative enzymes | Acyl-CoA thioesterases (ACOTs), including ACOT8, ACOT12, and metallo-beta-lactamase domain-containing protein 2 (MBLAC2) |
| Subcellular locations | Peroxisomes, mitochondria, cytosol, and brain tissue |
| Regulation | Influenced by peroxisomal proliferators, dietary status, and diabetes |
What Is GO:0016289?
According to the Gene Ontology, acyl-CoA hydrolase activity (GO:0016289) is defined as the catalysis of the reaction: an acyl-CoA + H2O = a carboxylate + CoA + H+. In simpler terms, it is an enzymatic activity that cleaves the thioester bond between a fatty acid and coenzyme A, freeing the fatty acid and CoA. This activity is synonymous with CoA hydrolase activity and is classified under the molecular_function ontology aspect.
Why Is acyl-CoA hydrolase activity Important in Cell Biology?
Acyl-CoA hydrolase activity is essential for maintaining cellular lipid homeostasis and energy balance. By controlling the availability of acyl-CoA, these enzymes influence fatty acid oxidation, lipid biosynthesis, and cell signaling. Dysregulation of this activity has been linked to cancer, metabolic disorders, and osteoarthritis, making it a promising target for therapeutic intervention.
• Regulates intracellular acyl-CoA pools, which are central to fatty acid metabolism and energy production.
• Modulates glycerolipid biosynthesis and suppresses YAP-mediated hepatocarcinogenesis through ACOT12.
• Promotes gemcitabine resistance in pancreatic ductal adenocarcinoma via ACOT8-mediated lipid metabolism and antiferroptotic activity.
• Influences brain lipid metabolism through long-chain acyl-CoA hydrolase activity.
• Is induced by peroxisomal proliferators in liver peroxisomes and mitochondria.
• Altered by dietary status and diabetes in aortic tissue, linking it to atherosclerosis.
• Plays a role in osteoarthritis biology, as reviewed in 2022.
• MBLAC2 exhibits acyl-CoA hydrolase activity and is S-palmitoylated, suggesting a role in protein lipidation.
• Provides a mechanism for fine-tuning lipid signaling molecules and membrane composition.
• Represents a potential therapeutic target for metabolic diseases and cancer.
Molecular Mechanism of acyl-CoA hydrolase activity
Substrate Recognition and Binding
In simple terms: The enzyme grabs onto an acyl-CoA molecule, which is a fatty acid attached to coenzyme A.
Acyl-CoA hydrolases recognize their substrates through a conserved thioesterase domain that accommodates the acyl chain and the CoA moiety. The binding pocket typically includes a catalytic triad or a metallo-beta-lactamase fold, as seen in MBLAC2, which is S-palmitoylated and exhibits acyl-CoA hydrolase activity. The specificity for different acyl chain lengths varies among enzymes; for example, long-chain acyl-CoA hydrolase in the brain prefers long-chain acyl-CoAs.
Catalytic Hydrolysis
In simple terms: Water is used to break the bond between the fatty acid and CoA, releasing both parts.
The hydrolysis reaction proceeds via nucleophilic attack of a water molecule on the thioester carbonyl, leading to the release of a free carboxylate (fatty acid), CoA, and a proton. This reaction is energetically favorable and does not require ATP. The catalytic mechanism often involves a serine or cysteine residue that forms an acyl-enzyme intermediate, which is subsequently hydrolyzed.
Subcellular Localization and Compartmentalization
In simple terms: These enzymes work in different parts of the cell, such as peroxisomes and mitochondria.
Acyl-CoA hydrolase activity is found in multiple cellular compartments, including peroxisomes, mitochondria, and the cytosol. In rat brown adipose tissue, the activity is present in peroxisomes, while in liver, peroxisomal proliferators enhance activity in both peroxisomes and mitochondria. This compartmentalization allows for distinct regulation of acyl-CoA pools in different metabolic pathways.
Regulation by Nutritional and Hormonal Signals
In simple terms: What you eat and hormones like insulin can change how active these enzymes are.
Acyl-CoA hydrolase activity is responsive to dietary status and diabetes. In aortic tissue, dietary changes and diabetes influence acyl-CoA hydrolase activity. Additionally, peroxisomal proliferators such as clofibrate increase long-chain acyl-CoA hydrolase activity in liver peroxisomes and mitochondria. These regulatory mechanisms help adapt lipid metabolism to changing physiological conditions.
Role in Lipid Metabolism and Signaling
In simple terms: By breaking down acyl-CoAs, these enzymes control the building blocks for fats and signaling molecules.
The hydrolysis of acyl-CoAs by acyl-CoA hydrolases directly impacts glycerolipid biosynthesis, as shown for ACOT12, which limits this pathway to suppress YAP-mediated hepatocarcinogenesis. In pancreatic cancer, ACOT8 regulates lipid metabolism and antiferroptotic activity, contributing to gemcitabine resistance. Thus, acyl-CoA hydrolase activity is a key node in lipid signaling and metabolic reprogramming.
Key Genes Involved in GO:0016289 acyl-CoA hydrolase activity
The following genes encode enzymes with acyl-CoA hydrolase activity or are directly associated with this function, as supported by the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ACOT12 | Acyl-CoA thioesterase 12; hydrolyzes acyl-CoAs to limit glycerolipid biosynthesis | Suppresses YAP-mediated hepatocarcinogenesis; potential tumor suppressor in liver cancer |
| ACOT8 | Acyl-CoA thioesterase 8; regulates lipid metabolism and ferroptosis | Induces gemcitabine resistance in pancreatic ductal adenocarcinoma |
| MBLAC2 | Metallo-beta-lactamase domain-containing protein 2; exhibits acyl-CoA hydrolase activity | S-palmitoylated; may link acyl-CoA hydrolysis to protein lipidation |
| ACOT1 | Acyl-CoA thioesterase 1; cytosolic long-chain acyl-CoA hydrolase | Involved in lipid metabolism; not directly cited in verified list but related to ACOT family |
| ACOT2 | Acyl-CoA thioesterase 2; mitochondrial long-chain acyl-CoA hydrolase | Related to ACOT family; not directly cited in verified list |
| ACOT3 | Acyl-CoA thioesterase 3; peroxisomal acyl-CoA hydrolase | Related to ACOT family; not directly cited in verified list |
| ACOT4 | Acyl-CoA thioesterase 4; peroxisomal acyl-CoA hydrolase | Related to ACOT family; not directly cited in verified list |
| ACOT5 | Acyl-CoA thioesterase 5; peroxisomal acyl-CoA hydrolase | Related to ACOT family; not directly cited in verified list |
| ACOT6 | Acyl-CoA thioesterase 6; peroxisomal acyl-CoA hydrolase | Related to ACOT family; not directly cited in verified list |
| ACOT7 | Acyl-CoA thioesterase 7; brain-specific long-chain acyl-CoA hydrolase | Long-chain acyl-CoA hydrolase in the brain |
| ACOT9 | Acyl-CoA thioesterase 9; mitochondrial acyl-CoA hydrolase | Related to ACOT family; not directly cited in verified list |
| ACOT11 | Acyl-CoA thioesterase 11; also known as THEM1 | Related to ACOT family; not directly cited in verified list |
| ACOT13 | Acyl-CoA thioesterase 13; also known as THEM2 | Related to ACOT family; not directly cited in verified list |
| BTHD | Brown adipose tissue acyl-CoA hydrolase | Presence of acyl-CoA hydrolase in rat brown-adipose-tissue peroxisomes |
| Peroxisomal proliferator-induced enzyme | Long-chain acyl-CoA hydrolase induced by peroxisomal proliferators | Enhancement of activity in peroxisomes and mitochondria of rat liver |
| Aortic acyl-CoA hydrolase | Acyl-CoA hydrolase activity in aorta | Influenced by dietary status and diabetes |
| Osteoarthritis-related acyl-CoA hydrolase | Acyl-CoA hydrolase activity in cartilage | Reviewed in osteoarthritis year in review 2022 |
How Is acyl-CoA hydrolase activity Regulated?
Acyl-CoA hydrolase activity is regulated at multiple levels. Peroxisomal proliferators such as clofibrate enhance long-chain acyl-CoA hydrolase activity in rat liver peroxisomes and mitochondria. Dietary status and diabetes also modulate aortic acyl-CoA hydrolase activity, indicating nutritional and hormonal control. Additionally, post-translational modifications such as S-palmitoylation may regulate the activity or localization of enzymes like MBLAC2. In cancer, the expression and activity of ACOT12 and ACOT8 are linked to specific signaling pathways, including YAP-mediated transcription and ferroptosis regulation.
acyl-CoA hydrolase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ACOT12 | Hepatocellular carcinoma; suppression of YAP-mediated hepatocarcinogenesis | Knockout mouse or liver-specific overexpression; CRISPR KO in HepG2 cells |
| ACOT8 | Pancreatic ductal adenocarcinoma; gemcitabine resistance | CRISPR KO in pancreatic cancer cell lines; xenograft models |
| MBLAC2 | Protein lipidation and potential neurological disorders | Point mutation of S-palmitoylation site; knock-in mice |
| ACOT7 | Brain lipid metabolism; neurological disorders | Brain-specific knockout; overexpression in neuronal cultures |
| Aortic acyl-CoA hydrolase | Atherosclerosis; diabetes-associated vascular changes | Diet-induced diabetes models; aortic tissue analysis |
Acyl-CoA Hydrolase Activity in Cancer
ACOT12 suppresses YAP-mediated hepatocarcinogenesis by limiting glycerolipid biosynthesis, suggesting that loss of acyl-CoA hydrolase activity can promote liver cancer. In pancreatic ductal adenocarcinoma, ACOT8 induces gemcitabine resistance through regulation of lipid metabolism and antiferroptotic activity. These findings highlight the dual roles of acyl-CoA hydrolases in cancer, where they can act as tumor suppressors or promote chemoresistance depending on context.
Acyl-CoA Hydrolase Activity in Metabolic and Cardiovascular Disorders
Altered acyl-CoA hydrolase activity has been observed in atherosclerosis, with dietary status and diabetes influencing aortic acyl-CoA hydrolase activity. Peroxisomal proliferators enhance activity in liver peroxisomes and mitochondria, linking these enzymes to lipid-lowering responses. In brown adipose tissue, acyl-CoA hydrolase activity in peroxisomes may contribute to thermogenesis and energy expenditure.
Acyl-CoA Hydrolase Activity in Osteoarthritis and Brain Function
Osteoarthritis year in review 2022 highlights the importance of lipid metabolism in joint biology, where acyl-CoA hydrolase activity may influence cartilage homeostasis. In the brain, long-chain acyl-CoA hydrolase activity is critical for neuronal lipid metabolism, and its dysregulation could contribute to neurological disorders. MBLAC2, an acyl-CoA hydrolase, is S-palmitoylated, suggesting a role in protein trafficking and signaling in the nervous system.
From acyl-CoA hydrolase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of ACOT12 promote liver cancer? | ACOT12 knockout mouse or CRISPR KO in hepatocytes |
| Does ACOT8 mediate gemcitabine resistance? | ACOT8 knockout in pancreatic cancer cells followed by drug treatment |
| How does S-palmitoylation regulate MBLAC2 activity? | Point mutation of cysteine residue in MBLAC2; knock-in cells |
| What is the role of ACOT7 in brain lipid metabolism? | Brain-specific ACOT7 knockout mouse |
| Can overexpression of ACOT12 suppress tumor growth? | ACOT12 overexpression in liver cancer cell lines and xenografts |
| How does diet affect aortic acyl-CoA hydrolase activity? | Dietary intervention in diabetic animal models |
How to Study the acyl-CoA hydrolase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Acyl-CoA hydrolase activity assay | Enzymatic hydrolysis of acyl-CoA to CoA | Measuring activity in cell lysates or purified enzymes |
| Lipidomics (LC-MS) | Acyl-CoA and free fatty acid levels | Profiling metabolic changes after gene knockout |
| CRISPR-Cas9 knockout | Loss of gene function | Studying causal role of ACOT genes in cancer |
| CRISPR point mutation | Specific amino acid changes | Dissecting catalytic residues or regulatory sites |
| Knock-in reporter | Endogenous protein localization and dynamics | Tracking ACOT expression in live cells |
| Overexpression | Gain of function | Testing tumor suppressor or oncogenic potential |
| Co-immunoprecipitation | Protein-protein interactions | Identifying ACOT binding partners |
| Fluorescence microscopy | Subcellular localization | Visualizing ACOTs in peroxisomes or mitochondria |
Enzymatic Activity Assays
Acyl-CoA hydrolase activity can be measured using spectrophotometric or fluorometric assays that monitor the release of CoA from acyl-CoA substrates. These assays are typically performed on cell lysates or purified enzyme preparations and can be adapted for high-throughput screening.
Lipidomics and Metabolomics
Mass spectrometry-based lipidomics and metabolomics allow comprehensive profiling of acyl-CoA species and their hydrolysis products. This approach can reveal how genetic perturbations of acyl-CoA hydrolases alter cellular lipid pools.
CRISPR-Cas9 Genome Editing
CRISPR-Cas9 knockout, point mutation, and knock-in strategies enable precise manipulation of genes encoding acyl-CoA hydrolases. These models are essential for establishing causal relationships between enzyme activity and disease phenotypes.
Protein Interaction and Localization Studies
Co-immunoprecipitation, proximity ligation, and fluorescence microscopy can determine the subcellular localization and interaction partners of acyl-CoA hydrolases. For example, S-palmitoylation of MBLAC2 can be studied using click chemistry or acyl-biotin exchange assays.
How CRISPR Can Be Used to Study GO:0016289 acyl-CoA hydrolase activity
Knockout
CRISPR knockout of acyl-CoA hydrolase genes such as ACOT12 or ACOT8 allows researchers to assess their loss-of-function phenotypes. For example, ACOT12 knockout can increase glycerolipid biosynthesis and activate YAP, promoting hepatocarcinogenesis. Similarly, ACOT8 knockout sensitizes pancreatic cancer cells to gemcitabine by disrupting lipid metabolism and ferroptosis.
Point Mutation
Point mutations can be introduced into catalytic residues or regulatory sites of acyl-CoA hydrolases to dissect their molecular mechanisms. For instance, mutating the S-palmitoylation site of MBLAC2 can reveal how this modification affects its acyl-CoA hydrolase activity and localization.
Knock-in
Knock-in of epitope tags or fluorescent reporters at endogenous loci enables real-time tracking of acyl-CoA hydrolase expression and localization. This approach can be used to study ACOT7 in brain tissue or ACOT12 in liver cells under physiological conditions.
Overexpression
Overexpression of acyl-CoA hydrolases such as ACOT12 can suppress tumor growth by limiting lipid biosynthesis. Conversely, overexpression of ACOT8 may confer chemoresistance. These models help establish sufficiency in disease pathways.
How EDITGENE Supports acyl-CoA hydrolase activity Research
Researchers studying acyl-CoA hydrolase activity-related genes often need to determine whether a candidate gene is causally involved in lipid metabolism, cancer, or metabolic disease. EDITGENE provides end-to-end CRISPR solutions to generate precisely engineered cell models, enabling rigorous functional validation of GO:0016289-associated genes.
Contact EDITGENE today to design your custom CRISPR model for acyl-CoA hydrolase activity research.
Frequently Asked Questions About acyl-CoA hydrolase activity
What is acyl-CoA hydrolase activity?
Acyl-CoA hydrolase activity (GO:0016289) is the enzymatic catalysis of the reaction: an acyl-CoA + H2O = a carboxylate + CoA + H+. It breaks down acyl-CoA molecules into free fatty acids and coenzyme A.
What genes are involved in acyl-CoA hydrolase activity?
Key genes include ACOT12, ACOT8, ACOT7, and MBLAC2, which encode enzymes with acyl-CoA hydrolase activity.
What is the function of ACOT12?
ACOT12 suppresses YAP-mediated hepatocarcinogenesis by limiting glycerolipid biosynthesis.
How does ACOT8 contribute to cancer?
ACOT8 induces gemcitabine resistance in pancreatic ductal adenocarcinoma via regulation of lipid metabolism and antiferroptotic activity.
Where is acyl-CoA hydrolase activity found in the cell?
It is found in peroxisomes, mitochondria, and the cytosol, depending on the specific enzyme.
What diseases are associated with acyl-CoA hydrolase activity?
It is linked to hepatocellular carcinoma, pancreatic cancer, atherosclerosis, diabetes, and osteoarthritis.
How is acyl-CoA hydrolase activity regulated?
It is regulated by peroxisomal proliferators, dietary status, diabetes, and post-translational modifications such as S-palmitoylation.
What is the role of acyl-CoA hydrolase in the brain?
Long-chain acyl-CoA hydrolase activity in the brain is important for neuronal lipid metabolism.
Can CRISPR be used to study acyl-CoA hydrolase activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are powerful tools to dissect the function of acyl-CoA hydrolase genes.
What methods measure acyl-CoA hydrolase activity?
Enzymatic activity assays, lipidomics, and metabolomics are commonly used to measure acyl-CoA hydrolase activity and its effects on lipid metabolism.
Conclusion
Acyl-CoA hydrolase activity (GO:0016289) is a fundamental enzymatic function that regulates lipid metabolism and cellular energy balance. Its dysregulation is implicated in cancer, metabolic disorders, and osteoarthritis, making it a compelling target for therapeutic development. Advances in CRISPR genome editing now allow precise interrogation of the genes encoding these enzymes, paving the way for novel insights and treatments. EDITGENE provides comprehensive CRISPR services to support this research.
References
- 1. He H et al.. 2023. Acyl-CoA thioesterase 12 suppresses YAP-mediated hepatocarcinogenesis by limiting glycerolipid biosynthesis.. Cancer Lett 565:216210 PMID: 37150501
- 2. Malgapo MIP et al.. 2021. Metallo-β-lactamase domain-containing protein 2 is S-palmitoylated and exhibits acyl-CoA hydrolase activity.. J Biol Chem 296:100106 PMID: 33219126
- 3. Li BR et al.. 2025. Acyl-CoA thioesterase 8 induces gemcitabine resistance via regulation of lipid metabolism and antiferroptotic activity in pancreatic ductal adenocarcinoma.. Acta Pharmacol Sin 46(6):1742-1756 PMID: 39939803
- 4. Yamada J. 2005. Long-chain acyl-CoA hydrolase in the brain.. Amino Acids 28(3):273-8 PMID: 15731883
- 5. Alexson SE et al.. 1989. The presence of acyl-CoA hydrolase in rat brown-adipose-tissue peroxisomes.. Biochem J 262(1):41-6 PMID: 2573347
- 6. Han S. 2022. Osteoarthritis year in review 2022: biology.. Osteoarthritis Cartilage 30(12):1575-1582 PMID: 36150676
- 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. Hashimoto S et al.. 1977. Influence of dietary status and diabetes on aortic acyl-CoA hydrolase activity.. Atherosclerosis 26(3):289-96 PMID: 139897
