GO:0160215 deacylase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0160215 deacylase activity describes the hydrolysis of an acyl group from a substrate, producing a free carboxylic acid and HX.
• The term covers a broad class of enzymes, including sirtuins (SIRT1-7) and other hydrolases that remove acetyl, succinyl, malonyl, glutaryl, and long-chain acyl marks [1, 4, 5].
• Deacylase activity is central to protein acylation homeostasis, influencing chromatin, metabolism, stress responses, and aging.
• Dysregulated deacylase activity is implicated in cancer, neurodegeneration, metabolic disorders, and inflammatory diseases [1, 4].
• Researchers study deacylase activity using continuous assays, bioluminescence assays, activity-based probes, and structural biology [2, 3, 6, 7, 8].
• CRISPR knockout, point-mutation, knock-in, and overexpression models enable causal dissection of deacylase function in disease [1, 4].
Description
Deacylase activity (GO:0160215) is a molecular function defined as the catalysis of the reaction R-CO-X + H2O = R-COOH + HX, i.e., the hydrolysis of an acyl group from a substrate molecule. This activity is essential for reversing protein acylation, a major post-translational modification that regulates enzyme activity, protein stability, and protein-protein interactions. The deacylase family includes NAD+-dependent sirtuins (SIRT1-7) and other hydrolases that remove acetyl, succinyl, malonyl, glutaryl, and long-chain fatty acyl groups from lysine residues [1, 4, 5]. Because acylation is dynamically controlled, deacylases act as key homeostatic regulators in transcription, metabolism, and stress responses. Researchers study deacylase activity to understand fundamental biology and to develop therapeutics for cancer, neurodegeneration, and metabolic diseases [1, 4]. For example, SIRT5 is a mitochondrial deacylase that removes succinyl, malonyl, and glutaryl groups, and its modulation is being explored for therapeutic benefit. SIRT6 exhibits deacylase activity with structural and enzymatic plasticity, influencing DNA repair and metabolism. The breadth of deacylase substrates and the diversity of enzymes involved make this GO term a focal point for chemical biology, structural biology, and functional genomics [2, 3, 6, 7, 8]. This article provides a research-grade overview of GO:0160215, covering its definition, mechanism, key genes, disease links, and experimental methods. All statements are based on published literature and the QuickGO definition [1-8].
deacylase activity At A Glance
| GO ID | GO:0160215 |
|---|---|
| GO term | deacylase activity |
| Ontology | molecular_function |
| Synonym | none |
| Definition | Catalysis of the reaction: R-CO-X + H2O = R-COOH + HX, hydrolysis of an acyl group or groups from a substrate molecule. |
| Major function | Removal of acyl groups from proteins and other substrates, reversing acylation and regulating substrate activity. |
| Enzyme classes | Includes NAD+-dependent sirtuins (SIRT1-7) and other hydrolases such as HDACs with deacylase activity. |
| Substrates | Acetyl, succinyl, malonyl, glutaryl, and long-chain acyl groups on lysine residues. |
| Biological context | Protein acylation homeostasis, chromatin regulation, metabolism, stress response, and aging. |
What Is GO:0160215?
GO:0160215 deacylase activity is a molecular function term describing the catalysis of the reaction R-CO-X + H2O = R-COOH + HX, which is the hydrolysis of an acyl group or groups from a substrate molecule. In simpler terms, it is the enzymatic removal of an acyl chemical group from a target molecule, often a protein, using water. This activity is distinct from other hydrolases because it specifically cleaves acyl groups, which can be acetyl, succinyl, malonyl, glutaryl, or longer fatty acyl chains [1, 4, 5].
Why Is deacylase activity Important in Cell Biology?
Deacylase activity is fundamentally important because it counteracts protein acylation, a pervasive post-translational modification that controls nearly every aspect of cell biology. By removing acyl groups, deacylases regulate the activity, localization, and stability of target proteins, thereby influencing gene expression, metabolic flux, and stress responses [1, 4]. Dysregulation of deacylase activity is linked to cancer, neurodegeneration, metabolic disorders, and inflammatory diseases, making these enzymes attractive drug targets [1, 4]. Moreover, deacylases such as sirtuins are central to aging and longevity research [1, 5].
• Regulates protein acylation homeostasis, a key post-translational modification.
• Controls chromatin structure and gene expression through histone deacylation [2, 4].
• Modulates metabolic pathways via mitochondrial deacylases like SIRT5.
• Influences DNA repair and genome stability through SIRT6 deacylase activity.
• Implicated in cancer, neurodegeneration, and metabolic diseases [1, 4].
• Target for therapeutic modulation with small molecules [1, 3].
• Essential for stress responses, including oxidative stress via peroxynitrite inhibition.
• Provides a mechanism for dynamic regulation of enzyme activity.
• Enables chemical biology tools for probing sirtuin function [6, 7, 8].
• Contributes to aging and longevity pathways [1, 5].
Molecular Mechanism of deacylase activity
Substrate recognition and binding
In simple terms: The enzyme first grabs the target molecule and its acyl group.
Deacylases recognize specific acyl-lysine marks on substrate proteins through a conserved catalytic pocket. For sirtuins, the pocket accommodates different acyl groups, with SIRT5 preferring succinyl, malonyl, and glutaryl groups, while SIRT1, SIRT2, SIRT3, and SIRT6 act on acetyl and longer acyl chains [1, 4, 5]. Structural studies reveal that the acyl chain length and charge determine binding affinity and catalytic efficiency. Activity-based probes have been developed to profile SIRT1 deacylase activity in complex proteomes.
Catalytic hydrolysis
In simple terms: Water is used to cut the acyl group off the substrate.
The catalytic mechanism involves hydrolysis of the acyl-enzyme intermediate or direct attack of water on the acyl-lysine bond, releasing a free carboxylic acid and the deacylated substrate. For sirtuins, the reaction is NAD+-dependent, consuming NAD+ and producing nicotinamide and 2'-O-acyl-ADP-ribose [1, 4]. The deacylase reaction is thus tightly linked to cellular energy status.
Cofactors and coenzymes
In simple terms: Some deacylases need a helper molecule called NAD+ to work.
NAD+ is an essential cofactor for sirtuin deacylases, and its availability regulates activity [1, 4]. Other deacylases may use metal ions or different cofactors. Peroxynitrite, a reactive nitrogen species, can inhibit sirtuin deacylase activity by modifying cysteine residues, linking oxidative stress to deacylase regulation.
Regulation of deacylase activity
In simple terms: The activity of these enzymes can be turned up or down by cellular signals.
Deacylase activity is regulated at multiple levels, including NAD+ availability, post-translational modifications, protein-protein interactions, and small-molecule inhibitors [1, 3]. For example, SIRT5 activity can be modulated by metabolic state and by synthetic ligands. Peroxynitrite inhibits sirtuin deacylase activity, providing a link to inflammation and oxidative stress. Continuous and bioluminescence assays have been developed to measure deacylase activity in real time, facilitating discovery of regulators [2, 7, 8].
Biological outcomes
In simple terms: Removing acyl groups changes how proteins work and what cells do.
Deacylation alters protein function, stability, and interactions, impacting transcription, metabolism, DNA repair, and stress responses. For instance, SIRT6 deacetylation of histones regulates chromatin and DNA repair. SIRT5-mediated desuccinylation modulates mitochondrial enzymes. These outcomes are context-dependent and are studied using knockout and knock-in models [1, 4].
Key Genes Involved in GO:0160215 deacylase activity
The following genes encode proteins with demonstrated deacylase activity (GO:0160215) or are directly involved in its regulation, as supported by published literature [1-8].
| Gene | Major Role | Research Relevance |
|---|---|---|
| SIRT1 | NAD+-dependent deacetylase and deacylase | Activity-based probes developed; roles in metabolism, aging, cancer. |
| SIRT2 | Deacetylase and deacylase | Regulates cell cycle and neurodegeneration; assay development [7, 8]. |
| SIRT3 | Mitochondrial deacetylase and deacylase | Metabolic regulation; bioluminescence assays [7, 8]. |
| SIRT4 | Mitochondrial deacylase | Metabolic and stress responses; assay development [7, 8]. |
| SIRT5 | Mitochondrial desuccinylase, demalonylase, glutarylase | Therapeutic target; activity modulation. |
| SIRT6 | Deacetylase and deacylase | Structural plasticity; DNA repair, aging. |
| SIRT7 | Deacetylase and deacylase | rRNA transcription, cancer; assay development [7, 8]. |
| HDAC1 | Histone deacetylase with deacylase activity | Chromatin regulation; continuous assays. |
| HDAC2 | Histone deacetylase with deacylase activity | Chromatin regulation; continuous assays. |
| HDAC3 | Histone deacetylase with deacylase activity | Chromatin regulation; continuous assays. |
| HDAC6 | Tubulin deacetylase | Cytoskeletal regulation; continuous assays. |
| HDAC8 | Histone deacetylase | Chromatin regulation; continuous assays. |
| SIRT5 (isoform) | Mitochondrial deacylase | Modulation by small molecules. |
| SIRT6 (isoform) | Deacylase | Enzymatic plasticity. |
| SIRT1 (isoform) | Deacylase | Probe development. |
| SIRT2 (isoform) | Deacylase | Bioluminescence assay [7, 8]. |
| SIRT3 (isoform) | Deacylase | Bioluminescence assay [7, 8]. |
How Is deacylase activity Regulated?
Deacylase activity is regulated by cellular NAD+ levels, oxidative stress, and small-molecule modulators [1, 3]. Peroxynitrite inhibits sirtuin deacylase activity, linking inflammation to deacylase dysfunction. Continuous and bioluminescence assays enable real-time monitoring of regulation [2, 7, 8]. Additionally, protein acylation itself is dynamically controlled by acyltransferases and deacylases, forming a reversible cycle.
deacylase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| SIRT1 | Cancer, neurodegeneration, aging | Knockout and overexpression cell models. |
| SIRT2 | Neurodegeneration, cancer | Point-mutation and knockout models [7, 8]. |
| SIRT5 | Cancer, metabolic disorders | Knockout and knock-in models. |
| SIRT6 | Cancer, aging, DNA repair defects | Knock-in and point-mutation models. |
| SIRT3 | Metabolic syndrome | Knockout and overexpression models [7, 8]. |
Cancer
Deacylases such as SIRT1, SIRT2, SIRT5, and SIRT6 are implicated in cancer through regulation of tumor suppressors, oncogenes, and metabolic pathways [1, 4, 5]. SIRT5 modulation is being explored as a therapeutic strategy. SIRT6 deacylase activity influences DNA repair and genomic stability, affecting cancer risk.
Neurodegeneration
SIRT2 deacylase activity is linked to neurodegeneration, and its inhibition is considered neuroprotective [7, 8]. SIRT1 deacylase activity also plays roles in neuronal survival and aging.
Metabolic disorders
Mitochondrial deacylases SIRT3, SIRT4, and SIRT5 regulate metabolic enzymes, and their dysfunction is associated with obesity, insulin resistance, and metabolic syndrome [1, 4, 7, 8].
Inflammatory and oxidative stress-related diseases
Peroxynitrite-mediated inhibition of sirtuin deacylase activity links oxidative stress to inflammatory diseases. Deacylase dysfunction may contribute to chronic inflammation and tissue damage.
From deacylase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of deacylase activity affect cell proliferation? | CRISPR knockout of SIRT5 in cancer cell lines. |
| How does a specific point mutation alter catalytic activity? | Point-mutation knock-in of SIRT6 catalytic residue. |
| What is the effect of deacylase overexpression on metabolism? | Overexpression of SIRT3 in metabolic cell models [7, 8]. |
| Can a tagged deacylase be used for interactome studies? | Tagged knock-in of SIRT1 with FLAG or HA. |
| Does deacylase activity modulate drug sensitivity? | Knockout and overexpression in isogenic cell lines [1, 4]. |
| What is the role of deacylase in oxidative stress? | Point-mutation of cysteine residues in SIRT2. |
How to Study the deacylase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Continuous deacylase assay | Real-time hydrolysis of acyl groups | High-throughput screening. |
| Bioluminescence assay | Sirtuin deacylase activity | Kinetic analysis and inhibitor testing [7, 8]. |
| Activity-based probes | Active deacylase enzymes | Proteome profiling. |
| Western blot with acyl-lysine antibodies | Levels of acylated proteins | Target engagement and cellular studies. |
| Mass spectrometry | Acyl modification sites | Global acylation analysis. |
| CRISPR knockout | Loss of deacylase function | Causal gene studies [1, 4]. |
| Site-directed mutagenesis | Effect of point mutations | Catalytic residue analysis. |
| Structural biology (X-ray/cryo-EM) | 3D structure of deacylase | Mechanism and drug design. |
Continuous deacylase activity assays
Continuous assays measure deacylase activity in real time by monitoring the release of product, such as free carboxylic acid or NAD+ consumption. These assays are suitable for high-throughput screening of inhibitors and activators.
Bioluminescence assays
Bioluminescence assays for lysine deacylase sirtuin activity provide sensitive detection of deacylation in cell lysates and live cells [7, 8]. They enable kinetic analysis and screening of modulators [7, 8].
Activity-based probes
Activity-based probes covalently label active deacylases, allowing profiling of enzyme activity in complex proteomes. These probes have been developed for SIRT1 and can be adapted for other deacylases.
Structural biology and enzymology
X-ray crystallography, cryo-EM, and kinetic studies reveal substrate binding, catalytic mechanism, and conformational changes. Such studies inform the design of selective inhibitors [1, 5].
How CRISPR Can Be Used to Study GO:0160215 deacylase activity
Knockout
CRISPR knockout of deacylase genes (e.g., SIRT5, SIRT6) eliminates enzyme activity, enabling studies of loss-of-function phenotypes in cancer, metabolism, and aging [1, 4, 5]. Knockout cell models are essential for validating target dependency and drug specificity.
Point Mutation
Point mutations in catalytic residues (e.g., SIRT6 H133Y) abolish deacylase activity while preserving protein structure, allowing separation of catalytic and non-catalytic functions. Such models are valuable for dissecting enzymatic versus scaffolding roles.
Knock-in
Knock-in of tagged or mutant deacylases (e.g., FLAG-SIRT1) enables affinity purification, imaging, and interactome studies. Knock-in of disease-associated variants helps assess their impact on deacylase activity [1, 4].
Overexpression
Overexpression of deacylases (e.g., SIRT3, SIRT5) in cell lines allows gain-of-function studies, including effects on metabolism, stress resistance, and drug response [1, 7, 8]. Overexpression models complement knockout studies for bidirectional analysis [1, 4].
How EDITGENE Supports deacylase activity Research
Researchers studying deacylase activity-related genes often need to determine whether a candidate gene is causally involved in a specific phenotype, and CRISPR-based models provide the most direct approach. EDITGENE offers a comprehensive suite of services to generate knockout, point-mutation, knock-in, and overexpression cell models, as well as CRISPR library screening and bioinformatics support.
Contact EDITGENE today to design your custom CRISPR model for deacylase activity research.
Frequently Asked Questions About deacylase activity
What is deacylase activity?
Deacylase activity (GO:0160215) is the catalysis of the reaction R-CO-X + H2O = R-COOH + HX, removing an acyl group from a substrate molecule.
What genes are involved in deacylase activity?
Key genes include SIRT1-7 and HDAC family members, which encode enzymes that remove acetyl, succinyl, malonyl, and other acyl groups [1, 4, 5].
What is the GO term for deacylase activity?
The Gene Ontology term is GO:0160215, under molecular_function.
How is deacylase activity measured?
It can be measured using continuous assays, bioluminescence assays, and activity-based probes [2, 6, 7, 8].
What diseases are linked to deacylase activity?
Dysregulated deacylase activity is linked to cancer, neurodegeneration, metabolic disorders, and inflammatory diseases [1, 3, 4, 5].
What is the role of SIRT5 deacylase activity?
SIRT5 is a mitochondrial deacylase that removes succinyl, malonyl, and glutaryl groups, and its modulation is explored for therapeutic benefit.
How does peroxynitrite affect deacylase activity?
Peroxynitrite inhibits sirtuin deacylase activity, linking oxidative stress to deacylase dysfunction.
What are the substrates of deacylase activity?
Substrates include acetyl, succinyl, malonyl, glutaryl, and long-chain acyl groups on lysine residues of proteins [1, 4, 5].
Can CRISPR be used to study deacylase activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are widely used to study deacylase function [1, 4, 5].
What is the difference between deacylase and deacetylase?
Deacylase activity is broader, encompassing removal of various acyl groups, while deacetylase specifically removes acetyl groups; many deacetylases also have deacylase activity [1, 4].
Conclusion
GO:0160215 deacylase activity is a fundamental molecular function that reverses protein acylation, impacting transcription, metabolism, stress responses, and disease. The diversity of deacylases, including sirtuins and HDACs, and their links to cancer, neurodegeneration, and metabolic disorders make them important research targets [1-8]. Advances in assays, probes, and CRISPR models continue to accelerate the field [2, 6, 7, 8]. EDITGENE provides comprehensive CRISPR services to support functional studies of deacylase activity, from knockout and point-mutation models to library screening and bioinformatics [1, 4, 5].
References
- 1. Fiorentino F et al.. 2022. Therapeutic Potential and Activity Modulation of the Protein Lysine Deacylase Sirtuin 5.. J Med Chem 65(14):9580-9606 PMID: 35802779
- 2. Zessin M et al.. 2023. Continuous Histone Deacylase Activity Assays.. Methods Mol Biol 2589:411-428 PMID: 36255640
- 3. Bohl K et al.. 2024. Inhibition of Sirtuin Deacylase Activity by Peroxynitrite.. Biochemistry 63(19):2463-2476 PMID: 39256054
- 4. Shang S et al.. 2022. Protein acylation: mechanisms, biological functions and therapeutic targets.. Signal Transduct Target Ther 7(1):396 PMID: 36577755
- 5. Wang ZA et al.. 2025. Structural and enzymatic plasticity of SIRT6 deacylase activity.. J Biol Chem 301(5):108446 PMID: 40147774
- 6. Goetz CJ et al.. 2020. Development of activity-based probes for the protein deacylase Sirt1.. Bioorg Chem 104:104232 PMID: 32911193
- 7. Van Scoyk AN et al.. 2024. Bioluminescence assay of lysine deacylase sirtuin activity.. Cell Chem Biol 31(11):2002-2014.e4 PMID: 39515338
- 8. Van Scoyk AN et al.. 2023. Bioluminescence Assay of Lysine Deacylase Sirtuin Activity.. bioRxiv PMID: 37645727
