GO:0004143 ATP-dependent diacylglycerol kinase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0004143 describes ATP-dependent diacylglycerol kinase activity, the catalysis of a 1,2-diacyl-sn-glycerol plus ATP to a 1,2-diacyl-sn-glycero-3-phosphate plus ADP and H+.
• Diacylglycerol kinases (DGKs) are lipid kinases that phosphorylate diacylglycerol (DAG) to produce phosphatidic acid (PA), a key lipid second messenger.
• The catalytic mechanism requires ATP as the phosphate donor and is dependent on membrane lipid bilayers for proper enzyme structure and function.
• DGK substrate acyl chain specificity is influenced by membrane morphology, linking enzyme activity to membrane curvature and lipid packing.
• Chemical proteomics and activity-based profiling have revealed the ligand binding landscape of DGKs, enabling development of selective inhibitors.
• Dysregulation of DGK activity is implicated in metabolic and signaling disorders, making it a target for therapeutic intervention and CRISPR-based disease modeling.
Description
ATP-dependent diacylglycerol kinase activity (GO:0004143) is a molecular function that catalyzes the phosphorylation of diacylglycerol (DAG) to phosphatidic acid (PA) using ATP as the phosphate donor. This reaction is central to lipid signaling, as it terminates DAG-mediated signals and simultaneously generates PA, a lipid second messenger involved in diverse cellular processes. The enzyme responsible, diacylglycerol kinase (DGK), exists in multiple isoforms with distinct regulatory properties and tissue distributions. Understanding this activity is critical for researchers studying lipid metabolism, signal transduction, and related diseases. The reaction is defined as: a 1,2-diacyl-sn-glycerol + ATP = a 1,2-diacyl-sn-glycero-3-phosphate + ADP + H+. DGK activity has been studied for decades, with early work identifying ATP-dependent inactivation of related enzymes in adipose tissue. Recent structural and chemical proteomic studies have provided insights into the membrane-bound conformation of DGK and its ligand binding landscape, facilitating the design of selective modulators. This article synthesizes current knowledge on GO:0004143, covering its mechanism, key genes, disease relevance, and research methodologies including CRISPR-based models.
ATP-dependent diacylglycerol kinase activity At A Glance
| GO ID | GO:0004143 |
|---|---|
| GO term | ATP-dependent diacylglycerol kinase activity |
| Ontology | molecular_function |
| Synonym | DGK activity; diacylglycerol kinase activity; diacylglycerol kinase activity (ATP); diglyceride kinase activity |
| Major function | Phosphorylation of diacylglycerol to phosphatidic acid using ATP |
| Reaction | a 1,2-diacyl-sn-glycerol + ATP = a 1,2-diacyl-sn-glycero-3-phosphate + ADP + H+ |
| Cofactor | ATP (required) |
| Localization | Membrane-associated, particularly in lipid bilayers |
| Substrate specificity | Influenced by membrane morphology and acyl chain composition |
What Is GO:0004143?
ATP-dependent diacylglycerol kinase activity (GO:0004143) is defined as the catalysis of the reaction: a 1,2-diacyl-sn-glycerol + ATP = a 1,2-diacyl-sn-glycero-3-phosphate + ADP + H+. In simpler terms, it is the enzyme activity that transfers a phosphate group from ATP to diacylglycerol, producing phosphatidic acid and ADP. This activity is synonymous with DGK activity, diacylglycerol kinase activity, diacylglycerol kinase activity (ATP), and diglyceride kinase activity. It is a molecular function classified under the Gene Ontology, and it requires ATP as a co-substrate and is typically associated with membrane lipid bilayers.
Why Is ATP-dependent diacylglycerol kinase activity Important in Cell Biology?
ATP-dependent diacylglycerol kinase activity is crucial for terminating diacylglycerol signaling and generating phosphatidic acid, a lipid second messenger that regulates cell growth, survival, and metabolism. Dysregulation of this activity has been linked to various pathological conditions, including metabolic disorders and cancer. Understanding the molecular mechanism and regulation of DGK is essential for developing therapeutic strategies targeting lipid signaling pathways. Moreover, the enzyme's dependence on membrane environment highlights the interplay between lipid composition and enzyme function, which is relevant to membrane biology and drug discovery.
• Terminates DAG-mediated signaling by converting DAG to PA.
• Generates PA, a lipid second messenger involved in cell proliferation and survival.
• Regulates metabolic pathways, as evidenced by ATP-dependent inactivation of related enzymes in adipose tissue.
• Implicated in cancer and metabolic diseases through altered lipid signaling.
• Target for chemical proteomics and inhibitor development.
• Membrane morphology influences substrate specificity, linking enzyme activity to membrane dynamics.
• Structural studies in lipid bilayers provide insights into conformational changes during catalysis.
• Activity-based profiling enables monitoring of DGK activity in complex proteomes.
• Potential role in cardiac function via regulation of Na+/H+ exchanger.
• CRISPR-based models can help dissect isoform-specific functions in disease.
What Happens During ATP-dependent diacylglycerol kinase activity?
Substrate Binding and Membrane Interaction
In simple terms: The enzyme attaches to the membrane and grabs its substrate, DAG.
Diacylglycerol kinase (DGK) binds to membrane lipid bilayers, where it interacts with its substrate, diacylglycerol (DAG). The membrane environment is critical for proper enzyme conformation and activity, as structural studies in lipid bilayers have shown. The enzyme's substrate acyl chain specificity is influenced by membrane morphology, such as curvature and lipid packing. This step ensures that DGK acts at the right place and time to modulate lipid signaling.
ATP-Dependent Phosphorylation
In simple terms: The enzyme uses ATP to add a phosphate group to DAG, making PA.
Upon binding DAG, DGK catalyzes the transfer of a phosphate group from ATP to the sn-3 position of DAG, yielding phosphatidic acid (PA) and ADP. This reaction is ATP-dependent and is the defining catalytic event of GO:0004143. The reaction also releases a proton (H+). The catalytic mechanism involves conserved residues that coordinate ATP and stabilize the transition state, as inferred from structural and biochemical studies.
Product Release and Signaling
In simple terms: The products PA and ADP are released, and PA goes on to send signals.
After catalysis, phosphatidic acid (PA) and ADP are released. PA acts as a lipid second messenger, recruiting effector proteins to membranes and regulating various cellular processes. The release of PA is a key step in terminating DAG signaling and initiating PA-mediated pathways. The balance between DAG and PA levels is crucial for cellular homeostasis.
Regulation by Membrane Morphology
In simple terms: The shape of the membrane can affect how well the enzyme works.
Membrane morphology, including curvature and lipid composition, modulates DGK substrate specificity and activity. For example, DGKα preferentially phosphorylates certain DAG species depending on the membrane environment. This regulation ensures that DGK activity is tuned to specific cellular contexts, such as during vesicle trafficking or signal transduction.
Inactivation and Turnover
In simple terms: The enzyme can be turned off or degraded when no longer needed.
DGK activity can be regulated by post-translational modifications, such as phosphorylation, and by interaction with other proteins. Early studies described ATP-dependent inactivation of diacylglycerol acyltransferase, a related enzyme, suggesting that ATP levels can influence enzyme stability. Turnover of DGK is also controlled by proteasomal degradation, ensuring tight regulation of lipid signaling.
Key Genes Involved in GO:0004143 ATP-dependent diacylglycerol kinase activity
The following genes encode diacylglycerol kinases and related proteins that exhibit ATP-dependent diacylglycerol kinase activity or regulate it.
| Gene | Major Role | Research Relevance |
|---|---|---|
| DGKA | Encodes diacylglycerol kinase alpha, a member of the DGK family | Studied for its role in T-cell signaling and cancer |
| DGKB | Encodes diacylglycerol kinase beta | Implicated in neuronal signaling and metabolic regulation |
| DGKG | Encodes diacylglycerol kinase gamma | Expressed in retina and brain; potential role in vision |
| DGKD | Encodes diacylglycerol kinase delta | Involved in glucose metabolism and insulin signaling |
| DGKE | Encodes diacylglycerol kinase epsilon | Mutations linked to atypical hemolytic uremic syndrome |
| DGKH | Encodes diacylglycerol kinase eta | Associated with bipolar disorder and lithium response |
| DGKI | Encodes diacylglycerol kinase iota | Expressed in brain; potential role in synaptic plasticity |
| DGKQ | Encodes diacylglycerol kinase theta | Involved in platelet activation and immune signaling |
| DGKZ | Encodes diacylglycerol kinase zeta | Regulates cell cycle and cytoskeleton dynamics |
| DGAT1 | Diacylglycerol acyltransferase 1; related enzyme | Subject of ATP-dependent inactivation studies |
| DGAT2 | Diacylglycerol acyltransferase 2; related enzyme | Involved in triglyceride synthesis |
| PRKCA | Protein kinase C alpha; downstream effector of DAG | DAG signaling target; interacts with DGK pathway |
| PRKCB | Protein kinase C beta | Mediates DAG signaling; potential crosstalk with DGK |
| SLC9A1 | Na+/H+ exchanger 1; regulated by DGK-derived PA | Cardiac function and disease |
| AGPAT1 | Acylglycerol-3-phosphate O-acyltransferase 1; PA metabolism | Downstream of DGK in lipid synthesis |
| PLD1 | Phospholipase D1; produces PA alternatively | Crosstalk with DGK in PA generation |
| PIP5K1A | Phosphatidylinositol-4-phosphate 5-kinase; uses PA | Links DGK to phosphoinositide signaling |
How Is ATP-dependent diacylglycerol kinase activity Regulated?
ATP-dependent diacylglycerol kinase activity is regulated at multiple levels. Membrane lipid composition and morphology directly influence substrate specificity and catalytic efficiency. Post-translational modifications, such as phosphorylation, can modulate DGK activity and localization. Additionally, ATP availability affects enzyme function, as demonstrated by ATP-dependent inactivation of related diacylglycerol acyltransferases. Chemical proteomic studies have revealed that DGKs bind various ligands, including inhibitors, which can be used to probe regulation. The activity is also subject to feedback regulation by downstream products like phosphatidic acid, which can affect enzyme recruitment and activity.
ATP-dependent diacylglycerol kinase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| DGKA | T-cell leukemia, cancer proliferation | DGKA knockout T-cell lines; xenograft models |
| DGKE | Atypical hemolytic uremic syndrome | DGKE knockout endothelial cells; mouse models |
| DGKH | Bipolar disorder | DGKH knockout neurons; behavioral studies |
| DGKQ | Platelet activation disorders | DGKQ knockout platelets; thrombosis models |
| DGKZ | Cell cycle dysregulation | DGKZ knockout cancer cell lines; proliferation assays |
Cancer and Cell Proliferation
Altered DGK activity has been observed in various cancers, where it contributes to sustained proliferation and survival signaling. DGKα, in particular, is implicated in T-cell leukemia and solid tumors, making it a potential therapeutic target. Chemical proteomic profiling has identified inhibitors that selectively modulate DGK activity, offering leads for anticancer drug development.
Metabolic Disorders
DGK activity is linked to metabolic regulation, as early studies showed ATP-dependent inactivation of diacylglycerol acyltransferase in adipose tissue, affecting triglyceride synthesis. Dysregulation of DGK isoforms may contribute to obesity, insulin resistance, and type 2 diabetes. Targeting DGK activity could provide therapeutic benefits in metabolic diseases.
Neurological and Psychiatric Disorders
DGK isoforms are highly expressed in the brain, where they regulate synaptic signaling and plasticity. DGKH has been associated with bipolar disorder, and DGKI with cognitive functions. Modulating DGK activity may offer strategies for treating neuropsychiatric conditions.
Cardiovascular and Renal Diseases
DGK-derived phosphatidic acid regulates the cardiac Na+/H+ exchanger, influencing cardiac function and disease. Mutations in DGKE are linked to atypical hemolytic uremic syndrome, a renal disorder. These findings highlight the importance of DGK in cardiovascular and renal physiology.
From ATP-dependent diacylglycerol kinase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does DGKA loss affect T-cell signaling? | DGKA knockout Jurkat cells via CRISPR |
| What is the effect of DGKE mutation on endothelial function? | DGKE point-mutation knock-in in HUVECs |
| How does DGKH variant affect neuronal signaling? | DGKH knock-in in iPSC-derived neurons |
| Can DGKZ overexpression alter cell cycle? | DGKZ overexpression in HeLa cells |
| What is the role of DGK isoforms in lipid metabolism? | DGK knockout mouse models |
| How does membrane morphology affect DGK activity? | In vitro lipid bilayer assays with purified DGK |
How to Study the ATP-dependent diacylglycerol kinase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Activity-based protein profiling | Enzyme activity in proteomes | Profiling DGK activity in cell lysates |
| Chemical proteomics | Ligand binding landscape | Identifying DGK inhibitors |
| Structural biology (cryo-EM/NMR) | 3D structure in lipid bilayers | Understanding catalytic mechanism |
| Lipid kinase inhibitor profiling | Inhibitor selectivity | Drug discovery for DGK targets |
| Membrane morphology assays | Substrate specificity | Linking membrane shape to activity |
| ATP-dependent inactivation assays | Enzyme stability | Studying ATP effects on related enzymes |
| Kinase activity assays | Phosphorylation rates | Measuring DGK catalytic activity |
| CRISPR knockout screens | Gene function | Identifying DGK roles in disease |
Activity-Based Protein Profiling
Activity-based protein profiling (ABPP) using chemical proteomics and ATP acyl phosphates enables monitoring of DGK activity in complex proteomes. This method allows researchers to assess enzyme activity states and identify selective inhibitors.
Structural Biology in Lipid Bilayers
Structural studies of membrane diacylglycerol kinase in lipid bilayers provide insights into the enzyme's conformation and catalytic mechanism. Techniques such as solid-state NMR and cryo-EM can reveal how membrane environment affects activity.
Lipid Kinase Inhibitor Profiling
Chemical proteomics can deconstruct lipid kinase inhibitors, identifying their targets and selectivity. This approach is valuable for developing drugs that modulate DGK activity in disease models.
Membrane Morphology Assays
Assays using lipid vesicles of varying curvature and composition can determine how membrane morphology influences DGK substrate specificity. Such methods help link biophysical properties to enzyme function.
How CRISPR Can Be Used to Study GO:0004143 ATP-dependent diacylglycerol kinase activity
Knockout
CRISPR knockout of DGK genes (e.g., DGKA, DGKE) in cell lines or primary cells can elucidate their roles in lipid signaling and disease. For example, DGKA knockout in T-cells can reveal its function in immune responses.
Point Mutation
Introducing point mutations in DGK catalytic residues or regulatory domains via CRISPR can dissect their contribution to enzyme activity and substrate specificity. This approach is useful for modeling disease-associated variants.
Knock-in
Knock-in of tagged DGK (e.g., GFP) allows live-cell imaging and proteomic analysis of enzyme localization and interactions. This can reveal how DGK is recruited to specific membranes.
Overexpression
Overexpression of DGK isoforms using CRISPR activation or lentiviral vectors can study gain-of-function effects in lipid signaling and disease models. This helps identify isoform-specific functions.
How EDITGENE Supports ATP-dependent diacylglycerol kinase activity Research
Researchers studying ATP-dependent diacylglycerol kinase activity-related genes often need to determine whether a candidate gene is causally involved in lipid signaling, metabolic regulation, or disease. EDITGENE provides comprehensive CRISPR-based services to generate precise cell models for such investigations.
Contact EDITGENE today to design your custom CRISPR model for ATP-dependent diacylglycerol kinase activity research.
Related Products
| Product name | Cat.No. | Species | Gene ID | |
|---|---|---|---|---|
| DGKH Knockout HEK293 Cell Line | EDJ-KQ1047 | Human | 160851 | Details Get a Quote |
| DGKK Knockout HEK293 Cell Line | EDJ-KQ1050 | Human | 139189 | Details Get a Quote |
| DGKD Knockout HEK293 Cell Line | EDJ-KQ1692 | Human | 8527 | Details Get a Quote |
| DGKI Knockout HEK293 Cell Line | EDJ-KQ1693 | Human | 9162 | Details Get a Quote |
| DGKE Knockout HEK293 Cell Line | EDJ-KQ1695 | Human | 8526 | Details Get a Quote |
| DGKB Knockout HEK293 Cell Line | EDJ-KQ1696 | Human | 1607 | Details Get a Quote |
| DGKG Knockout HEK293 Cell Line | EDJ-KQ1697 | Human | 1608 | Details Get a Quote |
| DGKQ Knockout HEK293 Cell Line | EDJ-KQ1698 | Human | 1609 | Details Get a Quote |
| AGK Knockout HEK293 Cell Line | EDJ-KQ12308 | Human | 55750 | Details Get a Quote |
| DGKA Knockout HEK293 Cell Line | EDJ-KQ17867 | Human | 1606 | Details Get a Quote |
| DGKZ Knockout HEK293 Cell Line | EDJ-KQ17868 | Human | 8525 | Details Get a Quote |
| DGKA Knockout A-549 Cell Line | EDJ-KQ20151 | Human | 1606 | Details Get a Quote |
| AGK Knockout A-549 Cell Line | EDJ-KQ41139 | Human | 55750 | Details Get a Quote |
| AGK Knockout HCT 116 Cell Line | EDJ-KQ41140 | Human | 55750 | Details Get a Quote |
| AGK Knockout HeLa Cell Line | EDC90512 | Human | 55750 | Details Get a Quote |
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Frequently Asked Questions About ATP-dependent diacylglycerol kinase activity
What is ATP-dependent diacylglycerol kinase activity?
It is the enzyme activity that catalyzes the phosphorylation of diacylglycerol to phosphatidic acid using ATP, classified as GO:0004143.
What genes are involved in ATP-dependent diacylglycerol kinase activity?
Genes encoding diacylglycerol kinases (DGKs) such as DGKA, DGKB, DGKG, DGKD, DGKE, DGKH, DGKI, DGKQ, and DGKZ are involved.
What is the reaction catalyzed by ATP-dependent diacylglycerol kinase?
The reaction is: a 1,2-diacyl-sn-glycerol + ATP = a 1,2-diacyl-sn-glycero-3-phosphate + ADP + H+.
How is ATP-dependent diacylglycerol kinase activity regulated?
It is regulated by membrane lipid composition, post-translational modifications, ATP availability, and feedback from products like phosphatidic acid.
What diseases are associated with diacylglycerol kinase activity?
Dysregulation is linked to cancer, metabolic disorders, neurological conditions, and cardiovascular/renal diseases.
What methods are used to study ATP-dependent diacylglycerol kinase activity?
Methods include activity-based protein profiling, chemical proteomics, structural biology, and CRISPR screens.
Can CRISPR be used to study diacylglycerol kinase genes?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are powerful tools to dissect DGK gene functions.
What is the role of membrane morphology in DGK activity?
Membrane morphology influences DGK substrate acyl chain specificity and catalytic efficiency.
How does DGK activity affect cell signaling?
DGK terminates DAG signaling and generates PA, which regulates proliferation, survival, and metabolism.
What are the synonyms for ATP-dependent diacylglycerol kinase activity?
Synonyms include DGK activity, diacylglycerol kinase activity, diacylglycerol kinase activity (ATP), and diglyceride kinase activity.
Conclusion
ATP-dependent diacylglycerol kinase activity (GO:0004143) is a fundamental molecular function that regulates lipid signaling by converting diacylglycerol to phosphatidic acid. Its importance spans metabolic, neurological, and cardiovascular biology, with implications for cancer and other diseases. Advances in structural biology, chemical proteomics, and CRISPR-based models continue to illuminate the mechanisms and therapeutic potential of DGK enzymes. EDITGENE's comprehensive CRISPR services empower researchers to create precise cell models for studying this activity and its related genes.
References
- 1. Lau TE et al.. 1996. A protein tyrosine kinase associated with the ATP-dependent inactivation of adipose diacylglycerol acyltransferase.. Lipids 31(3):277-83 PMID: 8900457
- 2. Rodriguez MA et al.. 1992. Reversible ATP-dependent inactivation of adipose diacylglycerol acyltransferase.. Lipids 27(8):577-81 PMID: 1328797
- 3. Li J et al.. 2021. Structure of membrane diacylglycerol kinase in lipid bilayers.. Commun Biol 4(1):282 PMID: 33674677
- 4. Franks CE et al.. 2019. Activity-Based Kinome Profiling Using Chemical Proteomics and ATP Acyl Phosphates.. Curr Protoc Chem Biol 11(3):e72 PMID: 31483100
- 5. Bozelli JC Jr et al.. 2021. Membrane morphology determines diacylglycerol kinase α substrate acyl chain specificity.. FASEB J 35(6):e21602 PMID: 33977628
- 6. Franks CE et al.. 2017. The Ligand Binding Landscape of Diacylglycerol Kinases.. Cell Chem Biol 24(7):870-880.e5 PMID: 28712745
- 7. Wakabayashi S et al.. 2013. Regulation of the cardiac Na⁺/H⁺ exchanger in health and disease.. J Mol Cell Cardiol 61:68-76 PMID: 23429007
- 8. McCloud RL et al.. 2018. Deconstructing Lipid Kinase Inhibitors by Chemical Proteomics.. Biochemistry 57(2):231-236 PMID: 29155586
