GO:0061698 protein deglutarylation: Mechanism, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0061698 protein deglutarylation is the enzymatic removal of a glutaryl group (CO-CH2-CH2-CH2-CO) from a lysine residue in a peptide or protein.
• SIRT5 is the principal mammalian deglutarylase, a mitochondrial sirtuin that reverses lysine glutarylation and other acyl modifications.
• Lysine glutarylation is a dynamic post-translational modification that regulates metabolic enzymes, including glutaryl-CoA dehydrogenase (GCDH) and carbamoyl phosphate synthetase 1 (CPS1).
• Deglutarylation controls glutaminolysis, lysine oxidation, and mitochondrial metabolism, linking the process to colorectal carcinogenesis and liver disease.
• ABHD11 catalyzes lipoyl deglutarylation, a distinct deglutarylation reaction that regulates mitochondrial and T cell metabolism.
• Dysregulated deglutarylation contributes to cancer, metabolic disorders, and viral pathogenesis, making SIRT5 and related enzymes therapeutic targets.
Description
Protein deglutarylation (GO:0061698) is the biological process that removes a glutaryl group (CO-CH2-CH2-CH2-CO) from a modified lysine residue in a peptide or protein. This reaction reverses lysine glutarylation, a recently discovered post-translational modification (PTM) that adds a five-carbon dicarboxyl acyl group to protein lysines. Because glutarylation alters protein charge, conformation, and catalytic activity, its removal by deglutarylation is a key regulatory event in cellular metabolism. The process is conserved from bacteria to humans and is mediated primarily by sirtuin 5 (SIRT5), an NAD+-dependent deacylase localized to mitochondria. Researchers study protein deglutarylation because it sits at the intersection of mitochondrial metabolism, epigenetics, and disease. SIRT5-catalyzed deglutarylation activates enzymes such as glutaryl-CoA dehydrogenase (GCDH) and carbamoyl phosphate synthetase 1 (CPS1), thereby promoting lysine oxidation and ammonia detoxification. In cancer, SIRT5-mediated deglutarylation enhances glutaminolysis and supports colorectal carcinogenesis. In liver disease, SIRT5-dependent post-translational modifications, including deglutarylation, are emerging as therapeutic targets. Additionally, ABHD11-mediated lipoyl deglutarylation regulates mitochondrial and T cell metabolism, expanding the biological scope of this process beyond SIRT5. This article provides a research-grade overview of GO:0061698, covering its definition, mechanism, key genes, disease relevance, and experimental methods. All statements are based on published literature and the QuickGO definition, with inline citations to verified PubMed references.
protein deglutarylation At A Glance
| GO ID | GO:0061698 |
|---|---|
| GO term | protein deglutarylation |
| Ontology | biological_process |
| Synonym | none |
| Major function | Removal of a glutaryl group from a lysine residue in a protein, reversing lysine glutarylation |
| Primary enzyme | SIRT5 (sirtuin 5), an NAD+-dependent mitochondrial deacylase |
| Additional enzyme | ABHD11, which catalyzes lipoyl deglutarylation |
| Key substrates | GCDH, CPS1, and other mitochondrial metabolic enzymes |
| Associated diseases | Colorectal cancer, liver disease, metabolic disorders |
What Is GO:0061698?
Protein deglutarylation (GO:0061698) is defined by QuickGO as the removal of a glutaryl group (CO-CH2-CH2-CH2-CO) from a residue in a peptide or protein. In practice, this means an enzyme cleaves the amide bond between the epsilon-amino group of a lysine residue and the glutaryl moiety, restoring the unmodified lysine. The reaction is a type of protein deacylation and is functionally opposed to lysine glutarylation, a PTM that adds the same glutaryl group. Deglutarylation is distinct from deglutamylation, demalonylation, and desuccinylation, although SIRT5 can catalyze several of these related deacylation reactions.
Why Is protein deglutarylation Important in Cell Biology?
Protein deglutarylation is important because it controls the activity of central metabolic enzymes and thereby influences mitochondrial function, glutaminolysis, and lysine oxidation. By removing glutaryl groups from lysines, SIRT5 and other deglutarylases reset the modification state of target proteins, allowing them to adopt active conformations or interact with partners. This regulatory layer is critical for cellular adaptation to metabolic stress and has been linked to cancer progression, liver disease, and viral infection. Understanding deglutarylation also provides a mechanistic basis for developing SIRT5 inhibitors and activators as therapeutics.
• Reverses lysine glutarylation, a PTM that alters protein charge and function.
• Activates GCDH by deglutarylation, promoting lysine oxidation in mitochondria.
• Supports glutaminolysis and colorectal carcinogenesis through SIRT5-dependent deglutarylation.
• Regulates mitochondrial metabolism and T cell function via ABHD11-mediated lipoyl deglutarylation.
• Contributes to liver disease pathogenesis and is a potential therapeutic target.
• Modulates viral replication, as shown for porcine deltacoronavirus M protein.
• Provides a mechanism for metabolic reprogramming in cancer cells.
• Is a target for small-molecule modulators of SIRT5.
• Links protein PTMs to cellular energy status via NAD+ dependence.
• Offers biomarkers and drug targets for metabolic and neoplastic diseases.
What Happens During protein deglutarylation?
Substrate recognition and binding
In simple terms: The enzyme finds and grabs the glutarylated protein.
Deglutarylases such as SIRT5 recognize target proteins bearing a glutaryl-lysine modification. SIRT5 binds NAD+ and the glutaryl-lysine side chain within its catalytic pocket, positioning the substrate for catalysis. Substrate specificity is determined by the enzyme's active site architecture and by protein-protein interactions that recruit SIRT5 to specific targets such as GCDH and CPS1.
NAD+-dependent catalysis
In simple terms: The enzyme uses NAD+ to cut the glutaryl group off the lysine.
SIRT5 catalyzes deglutarylation through an NAD+-dependent mechanism. The enzyme cleaves NAD+ and transfers the acetyl-like glutaryl group from the lysine to ADP-ribose, generating O-glutaryl-ADP-ribose and releasing the unmodified lysine. This reaction consumes NAD+ and thus couples deglutarylation to cellular energy status.
Product release and lysine restoration
In simple terms: The lysine is restored and the glutaryl group is released as a byproduct.
After catalysis, the deglutarylated protein is released with a free epsilon-amino group on the target lysine, restoring its positive charge and potential for other modifications. The glutaryl-ADP-ribose byproduct is released and can be further metabolized. This step is essential for reactivating enzymes such as GCDH, which requires a free lysine for catalytic activity.
Regulation by metabolic state
In simple terms: The process speeds up or slows down depending on the cell's energy and NAD+ levels.
Because SIRT5 requires NAD+, deglutarylation is sensitive to the cellular NAD+/NADH ratio and overall metabolic state. Conditions that alter NAD+ availability, such as caloric restriction or metabolic stress, can modulate deglutarylation rates. Additionally, SIRT5 expression levels and post-translational modifications of the enzyme itself regulate its activity.
Alternative deglutarylation by ABHD11
In simple terms: Another enzyme, ABHD11, can also remove glutaryl groups from a different target.
ABHD11 catalyzes lipoyl deglutarylation, removing a glutaryl group from the lipoyl moiety of mitochondrial enzymes. This reaction regulates mitochondrial and T cell metabolism independently of SIRT5. The existence of ABHD11-mediated deglutarylation demonstrates that GO:0061698 encompasses multiple enzymatic routes and substrate classes.
Key Genes Involved in GO:0061698 protein deglutarylation
The following genes and proteins are central to protein deglutarylation, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| SIRT5 | NAD+-dependent deglutarylase; removes glutaryl groups from lysine residues | Main enzyme for studying deglutarylation; target for inhibitors |
| GCDH | Glutaryl-CoA dehydrogenase; activated by SIRT5-mediated deglutarylation | Model substrate for lysine oxidation and deglutarylation assays |
| CPS1 | Carbamoyl phosphate synthetase 1; regulated by glutarylation/deglutarylation | Links deglutarylation to ammonia detoxification and urea cycle |
| ABHD11 | Lipoyl deglutarylase; regulates mitochondrial and T cell metabolism | Alternative deglutarylase for non-SIRT5 pathways |
| GLS | Glutaminase; supports glutaminolysis in cancer | SIRT5-deglutarylation axis promotes glutaminolysis in colorectal cancer |
| IDH2 | Isocitrate dehydrogenase 2; mitochondrial metabolic enzyme | Potential target of glutarylation/deglutarylation |
| SDHA | Succinate dehydrogenase complex flavoprotein subunit A | Mitochondrial enzyme subject to lysine acylation |
| PDHA1 | Pyruvate dehydrogenase E1 subunit alpha 1 | Metabolic enzyme with lysine glutarylation sites |
| ACAT1 | Acetyl-CoA acetyltransferase 1 | Mitochondrial enzyme regulated by acylation |
| HADHA | Hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha | Fatty acid oxidation enzyme with glutarylation sites |
| OTC | Ornithine transcarbamylase | Urea cycle enzyme potentially regulated by deglutarylation |
| SIRT3 | Mitochondrial deacetylase with overlapping substrate specificity | Comparative studies with SIRT5 |
| SIRT4 | Mitochondrial sirtuin with distinct acyl preferences | Context for SIRT5 specificity |
| NAMPT | Nicotinamide phosphoribosyltransferase; regulates NAD+ supply | Modulates SIRT5 activity by controlling NAD+ levels |
| PDCoV M protein | Viral membrane protein desuccinylated by SIRT5 | Example of SIRT5-mediated deacylation in viral infection |
| FOXO3a | Transcription factor linked to SIRT5-regulated metabolism | Potential downstream effector in cancer |
| GLUD1 | Glutamate dehydrogenase 1; glutaminolysis enzyme | Metabolic node in SIRT5-dependent cancer growth |
How Is protein deglutarylation Regulated?
Protein deglutarylation is regulated at multiple levels. The primary enzyme SIRT5 requires NAD+ as a co-substrate, so its activity is directly tied to the cellular NAD+/NADH ratio and metabolic status. SIRT5 expression can be transcriptionally regulated, and its catalytic activity is influenced by post-translational modifications and protein-protein interactions. In cancer, SIRT5-mediated deglutarylation is enhanced to support glutaminolysis, suggesting that oncogenic signaling pathways upregulate this process. In liver disease, SIRT5-dependent post-translational modifications, including deglutarylation, are modulated by metabolic stress and may be targeted therapeutically. Additionally, ABHD11 provides an alternative route for deglutarylation that is regulated independently of SIRT5.
protein deglutarylation and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| SIRT5 | Colorectal cancer; liver disease | SIRT5 knockout HCT116 xenograft; liver-specific SIRT5 KO mice |
| GCDH | Glutaric acidemia type I; lysine oxidation disorder | Gcdh-/- mouse model; SIRT5/GCDH double KO |
| CPS1 | Urea cycle disorder; hyperammonemia | CPS1 glutarylation site mutants in hepatocytes |
| ABHD11 | Mitochondrial metabolism; T cell function | ABHD11 knockout Jurkat or primary T cells |
| GLS | Cancer glutaminolysis | SIRT5 KO cancer cells with glutamine restriction |
Colorectal cancer
SIRT5 contributes to colorectal carcinogenesis by enhancing glutaminolysis in a deglutarylation-dependent manner. Elevated SIRT5 activity removes glutaryl groups from metabolic enzymes, promoting the use of glutamine as a carbon source and supporting tumor growth. This makes the deglutarylation pathway a potential target for colorectal cancer therapy.
Liver disease
SIRT5-mediated post-translational modifications, including deglutarylation, are emerging as a promising therapeutic approach to attenuate liver diseases. Dysregulated deglutarylation may affect mitochondrial metabolism and ammonia detoxification, contributing to hepatic dysfunction. Targeting SIRT5 or its substrates could offer new strategies for liver disease treatment.
Metabolic disorders and lysine oxidation
Deglutarylation of glutaryl-CoA dehydrogenase by SIRT5 promotes lysine oxidation in mice. Impaired deglutarylation could lead to accumulation of glutarylated proteins and metabolic imbalance, linking the process to inherited metabolic disorders such as glutaric acidemia type I. Understanding this pathway may inform therapeutic approaches for metabolic diseases.
Viral infection
SIRT5-mediated deacylation, including desuccinylation, regulates the porcine deltacoronavirus M protein and drives pexophagy to enhance viral proliferation. This demonstrates that deglutarylation-related enzymes can influence viral pathogenesis, although the specific role of deglutarylation in this context requires further study.
From protein deglutarylation-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does SIRT5 deglutarylate a specific substrate? | SIRT5 knockout cell line + substrate overexpression |
| What is the effect of a glutarylation site mutation? | Point-mutant knock-in of lysine-to-arginine or glutamine in target gene |
| How does deglutarylation affect metabolic flux? | SIRT5 KO or knock-in cells with 13C-glutamine tracing |
| Can a drug modulate deglutarylation? | SIRT5 inhibitor/activator treatment in wild-type and KO cells |
| What is the role of ABHD11 in deglutarylation? | ABHD11 knockout or overexpression in T cells |
| Does deglutarylation regulate viral replication? | SIRT5 KO cells infected with PDCoV |
How to Study the protein deglutarylation Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Glutaryl-lysine immunoprecipitation + LC-MS/MS | Identification and quantification of glutarylated proteins | Mapping deglutarylation targets in cells |
| In vitro deglutarylation assay | Enzymatic activity of SIRT5 or ABHD11 | Kinetic studies and inhibitor screening |
| 13C-glutamine tracing | Glutaminolysis flux | Cancer metabolism studies |
| 13C-lysine oxidation assay | Lysine oxidation rate | GCDH deglutarylation studies |
| Western blot with anti-glutaryl-lysine | Global glutarylation levels | SIRT5 KO validation |
| Mitochondrial respiration assay | OXPHOS function | ABHD11 and SIRT5 studies |
| T cell proliferation assay | Immune cell function | ABHD11 deglutarylation studies |
| Viral plaque assay | Viral replication | SIRT5-PDCoV interaction studies |
Proteomics and PTM enrichment
Glutaryl-lysine enrichment coupled with mass spectrometry is the primary method to identify deglutarylation targets and quantify site occupancy. Immunoprecipitation with anti-glutaryl-lysine antibodies followed by LC-MS/MS can map glutarylation sites on proteins such as GCDH and CPS1. Comparative proteomics between SIRT5 wild-type and knockout cells reveals substrates whose glutarylation increases upon loss of deglutarylation.
Enzymatic assays
In vitro deglutarylation assays use recombinant SIRT5, NAD+, and glutarylated peptide or protein substrates. Fluorescence-based or mass spectrometry-based readouts measure the release of glutaryl-ADP-ribose or the appearance of unmodified lysine. These assays are used to determine kinetic parameters and to screen for inhibitors.
Metabolic flux analysis
Stable isotope tracing with 13C-glutamine or 13C-lysine measures how deglutarylation affects glutaminolysis and lysine oxidation. SIRT5 knockout or overexpression models are used to link deglutarylation activity to metabolic flux and ATP production.
Cell-based imaging and functional assays
Fluorescently tagged SIRT5 or substrate proteins can be imaged to assess localization and deglutarylation-dependent changes. T cell proliferation and mitochondrial function assays are used to study ABHD11-mediated deglutarylation. Viral replication assays can test the impact of SIRT5 on PDCoV proliferation.
How CRISPR Can Be Used to Study GO:0061698 protein deglutarylation
Knockout
CRISPR knockout of SIRT5 or ABHD11 is used to abolish deglutarylation activity and assess downstream effects on glutarylation levels, metabolism, and disease phenotypes. SIRT5 knockout cell lines and mice show increased lysine glutarylation and altered glutaminolysis. Knockout of GCDH in combination with SIRT5 knockout can reveal substrate-specific effects.
Point Mutation
Point mutations at catalytic residues of SIRT5 (e.g., H158Y) or at glutarylation acceptor lysines in substrates (e.g., GCDH K335) can dissect the mechanism of deglutarylation. CRISPR-mediated knock-in of these point mutations allows study of deglutarylation in a physiological context without altering protein levels.
Knock-in
Knock-in of tagged SIRT5 (e.g., FLAG or GFP) enables affinity purification and localization studies of the deglutarylase. Knock-in of glutarylation-deficient or mimetic mutants of substrate proteins (e.g., CPS1 K-to-Q) can mimic constitutive deglutarylation or block it. These models are valuable for understanding site-specific regulation.
Overexpression
Overexpression of SIRT5 or ABHD11 via CRISPR activation or lentiviral delivery increases deglutarylation activity and can drive metabolic reprogramming. Overexpression of substrate proteins with glutarylation sites allows assessment of deglutarylation efficiency. These models are used to test whether increased deglutarylation promotes cancer or alters immune cell function.
How EDITGENE Supports protein deglutarylation Research
Researchers studying protein deglutarylation-related genes often need to determine whether a candidate gene is causally involved in a metabolic or disease phenotype. This requires precise genetic models that can knockout, mutate, tag, or overexpress the gene of interest in relevant cell types. EDITGENE provides end-to-end CRISPR services to generate such models, enabling rigorous investigation of GO:0061698 and its regulators.
Contact EDITGENE today to design your custom CRISPR model for protein deglutarylation research.
Frequently Asked Questions About protein deglutarylation
What is protein deglutarylation?
Protein deglutarylation (GO:0061698) is the removal of a glutaryl group (CO-CH2-CH2-CH2-CO) from a lysine residue in a protein, reversing lysine glutarylation.
What genes are involved in protein deglutarylation?
The main gene is SIRT5, which encodes an NAD+-dependent deglutarylase. ABHD11 also catalyzes lipoyl deglutarylation. Substrates include GCDH and CPS1.
Which enzyme performs protein deglutarylation?
SIRT5 is the principal mammalian deglutarylase, but ABHD11 can also remove glutaryl groups from lipoyl moieties.
What is the role of SIRT5 in deglutarylation?
SIRT5 uses NAD+ to remove glutaryl groups from lysine residues, thereby activating metabolic enzymes and promoting glutaminolysis and lysine oxidation.
How is protein deglutarylation linked to cancer?
SIRT5-mediated deglutarylation enhances glutaminolysis in a deglutarylation-dependent manner, contributing to colorectal carcinogenesis.
What diseases are associated with defective deglutarylation?
Colorectal cancer, liver disease, and metabolic disorders such as glutaric acidemia type I have been linked to altered deglutarylation.
How can I study protein deglutarylation in the lab?
Common methods include glutaryl-lysine immunoprecipitation, mass spectrometry, in vitro enzymatic assays, and metabolic flux analysis using SIRT5 knockout or mutant cells.
What is the difference between glutarylation and deglutarylation?
Glutarylation adds a glutaryl group to lysine, while deglutarylation removes it. SIRT5 reverses glutarylation.
Can CRISPR be used to study deglutarylation?
Yes, CRISPR knockout, knock-in, and point mutation models of SIRT5, ABHD11, and substrates are widely used to study deglutarylation.
Is SIRT5 a drug target for deglutarylation-related diseases?
Yes, SIRT5 inhibitors and activators are being developed as potential therapeutics for cancer and liver diseases.
Conclusion
Protein deglutarylation (GO:0061698) is a fundamental post-translational regulatory process that reverses lysine glutarylation and controls mitochondrial metabolism, glutaminolysis, and lysine oxidation. SIRT5 is the primary enzyme responsible, with ABHD11 providing an alternative route for lipoyl deglutarylation. Dysregulation of this process is implicated in colorectal cancer, liver disease, and metabolic disorders, making it a promising therapeutic target. Continued research using CRISPR models and advanced proteomics will further elucidate the mechanisms and disease relevance of deglutarylation.
References
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- 3. Tan M et al.. 2014. Lysine glutarylation is a protein posttranslational modification regulated by SIRT5.. Cell Metab 19(4):605-17 PMID: 24703693
- 4. 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
- 5. Wang YQ et al.. 2018. Sirtuin5 contributes to colorectal carcinogenesis by enhancing glutaminolysis in a deglutarylation-dependent manner.. Nat Commun 9(1):545 PMID: 29416026
- 6. Bhatt DP et al.. 2022. Deglutarylation of glutaryl-CoA dehydrogenase by deacylating enzyme SIRT5 promotes lysine oxidation in mice.. J Biol Chem 298(4):101723 PMID: 35157847
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- 8. Yue Y et al.. 2026. Sirtuin 5-mediated post-translational modifications as a promising therapeutic approach to attenuate liver diseases.. Eur J Pharmacol 1012:178498 PMID: 41421408
