GO:0042171 lysophosphatidic acid acyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0042171 describes the enzymatic activity that transfers an acyl group from acyl-CoA to lysophosphatidic acid (LPA), producing phosphatidic acid (PA), a central intermediate in glycerophospholipid biosynthesis.
• This activity is conserved from bacteria to plants and humans, with enzymes such as LPAATs, AGPATs, and CGI-58/ABHD5 catalyzing the reaction.
• LPAAT activity is critical for membrane biogenesis, lipid storage, and signaling, and its dysfunction is linked to diseases including cancer and metabolic disorders.
• Plant LPAATs, such as those in Brassica napus and Perilla frutescens, enhance seed oil accumulation and are targets for crop improvement.
• The enzyme exhibits substrate promiscuity in some organisms, allowing use of diverse acyl-CoAs, which has biotechnological implications.
• Research methods include CRISPR knockout/knock-in models, lipidomics, and enzyme assays to dissect LPAAT function in health and disease.
Description
Lysophosphatidic acid acyltransferase (LPAAT) activity, encoded by GO:0042171, is a fundamental enzymatic function in lipid metabolism. It catalyzes the acylation of lysophosphatidic acid (LPA) using acyl-CoA to generate phosphatidic acid (PA), a key intermediate in the synthesis of glycerophospholipids and triacylglycerols. This activity is essential for membrane lipid homeostasis and energy storage across all domains of life. In humans, LPAAT enzymes are involved in diverse physiological processes, and their dysregulation has been implicated in cancer, obesity, and neurological disorders. In plants, LPAATs contribute to seed oil production, making them targets for agricultural biotechnology. Understanding the molecular mechanisms, regulation, and disease relevance of LPAAT activity is therefore of broad scientific and therapeutic interest.
lysophosphatidic acid acyltransferase activity At A Glance
| GO ID | GO:0042171 |
|---|---|
| GO term | lysophosphatidic acid acyltransferase activity |
| Ontology | molecular_function |
| Synonym | LPAAT activity |
| Major function | Transfer of acyl group from acyl-CoA to lysophosphatidic acid to form phosphatidic acid |
| EC number | 2.3.1.51 |
| Reaction | acyl-CoA + 1-acyl-sn-glycerol 3-phosphate = CoA + 1,2-diacyl-sn-glycerol 3-phosphate |
| Substrates | Acyl-CoA and lysophosphatidic acid |
| Products | Phosphatidic acid and CoA |
What Is GO:0042171?
According to the Gene Ontology, GO:0042171 (lysophosphatidic acid acyltransferase activity) is defined as the catalysis of the transfer of acyl groups from an acyl-CoA to lysophosphatidic acid to form phosphatidic acid. This activity is synonymous with LPAAT activity and represents a key step in the Kennedy pathway for glycerophospholipid biosynthesis.
Why Is lysophosphatidic acid acyltransferase activity Important in Cell Biology?
LPAAT activity is a central node in lipid metabolism, controlling the synthesis of phosphatidic acid, which is a precursor for phosphatidylinositol, cardiolipin, and triacylglycerols. This activity influences membrane biogenesis, lipid signaling, and energy storage. In humans, LPAAT enzymes such as AGPAT2 are linked to congenital generalized lipodystrophy, and others are implicated in cancer progression and metabolic syndrome. In plants, LPAATs are critical for seed oil accumulation, with direct applications in biofuel and food industries. Thus, understanding LPAAT activity is vital for both basic cell biology and translational research.
• Essential for glycerophospholipid biosynthesis and membrane formation.
• Regulates lipid storage and adipocyte development; mutations cause lipodystrophy.
• Involved in cancer cell proliferation and survival through altered lipid signaling.
• Key determinant of seed oil content and composition in oilseed crops.
• Target for antibacterial drug discovery in Pseudomonas and other pathogens.
• Plays a role in thermophilic adaptation in bacteria like Thermus thermophilus.
• CGI-58/ABHD5 LPAAT activity links lipid droplets to neutral lipid storage disease.
• Provides a model for studying enzyme substrate promiscuity and evolution.
• Potential therapeutic target for metabolic disorders and cancer.
• Enables biotechnological production of tailored lipids and oils.
What Happens During lysophosphatidic acid acyltransferase activity?
Substrate Binding and Acyl Transfer
In simple terms: The enzyme grabs an acyl group from one molecule and attaches it to another.
LPAAT enzymes bind lysophosphatidic acid (LPA) and an acyl-CoA donor. The catalytic mechanism involves the transfer of the acyl group from acyl-CoA to the sn-2 position of LPA, forming phosphatidic acid and releasing CoA. This reaction is conserved across species, from bacteria to humans.
Role in the Kennedy Pathway
In simple terms: This step is part of a larger assembly line that builds membrane fats.
LPAAT activity constitutes the second step of the Kennedy pathway, converting LPA to PA. PA is then dephosphorylated to diacylglycerol (DAG) or converted to CDP-DAG for phospholipid synthesis. This pathway is essential for the production of phosphatidylcholine, phosphatidylethanolamine, and triacylglycerols.
Substrate Specificity and Promiscuity
In simple terms: Some versions of the enzyme are picky about their partners, while others are flexible.
LPAAT enzymes exhibit varying substrate specificities. For example, Thermus thermophilus LPAAT displays substrate promiscuity, utilizing a range of acyl-CoAs. In contrast, plant LPAATs often prefer unsaturated acyl-CoAs, influencing oil composition. This diversity has implications for enzyme engineering.
Cellular Compartmentalization
In simple terms: The reaction happens in different parts of the cell depending on the organism.
In plants, LPAATs are found in the plastid and endoplasmic reticulum, with distinct roles in seed oil biosynthesis. In humans, LPAAT isoforms localize to the endoplasmic reticulum and lipid droplets, where they participate in lipid storage and signaling. Compartmentalization ensures spatial and temporal control of lipid synthesis.
Key Genes Involved in GO:0042171 lysophosphatidic acid acyltransferase activity
The following genes and proteins are key players in lysophosphatidic acid acyltransferase activity, as supported by published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| AGPAT1 | LPAAT isoform in humans; catalyzes PA synthesis | Lipid metabolism and signaling |
| AGPAT2 | LPAAT isoform; mutations cause lipodystrophy | Disease modeling and therapeutic target |
| AGPAT3 | LPAAT isoform; involved in phospholipid remodeling | Cancer and metabolic studies |
| AGPAT4 | LPAAT delta; brain-specific functions | Neurodevelopment and lipid signaling |
| AGPAT5 | LPAAT epsilon; mitochondrial-associated | Energy metabolism |
| G0S2 | Intrinsic LPAAT activity; regulates lipolysis | Obesity and cancer |
| CGI-58/ABHD5 | CoA-dependent LPAAT; linked to Chanarin-Dorfman syndrome | Lipid storage diseases |
| LPAAT1 (Brassica napus) | Seed oil accumulation | Crop improvement |
| LPAAT2 (Brassica napus) | Seed oil accumulation | Crop improvement |
| LPAAT (Perilla frutescens) | Seed oil biosynthesis | Oilseed engineering |
| LPAAT (Pseudomonas fluorescens) | Membrane lipid synthesis | Antibacterial targets |
| Plastidial LPAAT (Brassica napus) | Plastid lipid synthesis | Oilseed rape oil quality |
| Thermus thermophilus LPAAT | Thermophilic membrane adaptation | Biotechnology and enzyme evolution |
How Is lysophosphatidic acid acyltransferase activity Regulated?
LPAAT activity is regulated at multiple levels. In humans, AGPAT2 expression is influenced by nutritional status and hormones, and its activity can be modulated by phosphorylation. CGI-58/ABHD5 LPAAT activity is activated by interaction with perilipins on lipid droplets, linking lipolysis to re-esterification. In plants, LPAAT genes are transcriptionally regulated during seed development, with expression peaks coinciding with oil accumulation. Additionally, substrate availability and membrane lipid composition can feedback-regulate LPAAT activity.
lysophosphatidic acid acyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| AGPAT2 | Congenital generalized lipodystrophy | Knockout mouse, patient-derived iPSCs |
| CGI-58/ABHD5 | Chanarin-Dorfman syndrome | Knock-in mouse models, lipid droplet assays |
| G0S2 | Cancer metabolism, obesity | Overexpression and knockout cell lines |
| AGPAT4 | Neurological disorders | Brain-specific knockout mice |
| LPAAT (Pseudomonas) | Bacterial infections | Bacterial knockout and inhibitor screens |
LPAAT activity in cancer
Altered LPAAT expression and activity have been observed in various cancers. AGPAT2 and AGPAT4 are overexpressed in some tumors, promoting cell proliferation and survival through increased phosphatidic acid signaling. G0S2, which possesses LPAAT activity, is implicated in cancer cell metabolism and apoptosis resistance. Targeting LPAAT enzymes is being explored as a therapeutic strategy.
Metabolic disorders and lipodystrophy
Mutations in AGPAT2 cause congenital generalized lipodystrophy, a severe metabolic disorder characterized by near-total absence of adipose tissue. CGI-58/ABHD5 mutations lead to Chanarin-Dorfman syndrome, a neutral lipid storage disease with ichthyosis, highlighting the importance of LPAAT activity in lipid homeostasis.
Infectious diseases
Bacterial LPAATs, such as those in Pseudomonas fluorescens, are essential for membrane phospholipid synthesis and are potential targets for new antibiotics. The unique substrate promiscuity of some bacterial LPAATs may offer selective inhibition opportunities.
From lysophosphatidic acid acyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Enzyme kinetics and substrate specificity | Purified recombinant LPAAT with mutant acyl-CoA |
| Role in seed oil accumulation | Plant knockout and overexpression lines |
| Lipid droplet dynamics and lipolysis | CRISPR knockout of CGI-58 in adipocytes |
| Membrane biogenesis in bacteria | Pseudomonas LPAAT deletion strains |
| Disease-associated mutations | Knock-in mouse models of AGPAT2 mutations |
| Thermostability and promiscuity | Directed evolution of Thermus LPAAT |
How to Study the lysophosphatidic acid acyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Radioenzymatic assay | LPAAT catalytic activity | Kinetic studies |
| LC-MS lipidomics | Phosphatidic acid and lipid species | Metabolic profiling |
| CRISPR knockout screen | Gene essentiality and modifiers | Functional genomics |
| Western blot | Protein expression levels | Validation of knockout/overexpression |
| Immunofluorescence | Subcellular localization | Organelle dynamics |
| qRT-PCR | mRNA expression | Transcriptional regulation |
| Enzyme-linked assay | CoA release | High-throughput screening |
Enzymatic assays for LPAAT activity
LPAAT activity is typically measured using radiolabeled or fluorescent acyl-CoA and LPA, followed by separation of products by thin-layer chromatography or HPLC. These assays allow determination of kinetic parameters and substrate specificity.
Lipidomics and mass spectrometry
Mass spectrometry-based lipidomics enables comprehensive profiling of phosphatidic acid and other lipids in cells or tissues with altered LPAAT expression. This method reveals changes in lipid species and pathways.
CRISPR-based genetic screens
Genome-wide CRISPR knockout or activation screens can identify genes that modulate LPAAT activity or compensate for its loss. Such screens are powerful for discovering synthetic lethal interactions.
Structural biology and modeling
X-ray crystallography and cryo-EM of LPAAT enzymes provide insights into catalytic mechanism and substrate binding. Homology modeling and molecular dynamics simulations complement experimental structures.
How CRISPR Can Be Used to Study GO:0042171 lysophosphatidic acid acyltransferase activity
Knockout
CRISPR knockout of LPAAT genes (e.g., AGPAT2, CGI-58) in cell lines or animal models abolishes enzymatic activity, enabling studies of lipid metabolism, membrane integrity, and disease phenotypes. Knockout models are essential for distinguishing isoform-specific functions.
Point Mutation
Introducing point mutations in catalytic residues or regulatory sites of LPAAT genes via CRISPR base editing or homology-directed repair allows precise dissection of enzyme mechanism and disease-associated variants. Such models help validate drug targets.
Knock-in
Knock-in of tagged LPAAT (e.g., GFP or FLAG) enables live-cell imaging and proteomic analysis of interacting partners. Knock-in of disease mutations (e.g., AGPAT2) recapitulates human pathology in model organisms.
Overexpression
CRISPR activation (CRISPRa) or cDNA overexpression of LPAAT genes increases enzymatic activity, useful for studying lipid accumulation, signaling, and resistance to therapies. Overexpression in plants boosts seed oil content.
How EDITGENE Supports lysophosphatidic acid acyltransferase activity Research
Researchers studying lysophosphatidic acid acyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in lipid metabolism, disease, or crop traits. EDITGENE provides a comprehensive suite of CRISPR services to accelerate this research, from gene knockout to precise point mutations and library screening.
Contact EDITGENE today to design your custom CRISPR model for lysophosphatidic acid acyltransferase activity research.
Frequently Asked Questions About lysophosphatidic acid acyltransferase activity
What is lysophosphatidic acid acyltransferase activity?
It is the enzymatic activity (GO:0042171) that transfers an acyl group from acyl-CoA to lysophosphatidic acid to form phosphatidic acid, a key step in lipid biosynthesis.
What genes are involved in lysophosphatidic acid acyltransferase activity?
Key genes include AGPAT1-5, G0S2, CGI-58/ABHD5 in humans, and LPAATs in plants and bacteria.
What diseases are associated with LPAAT activity?
Mutations in AGPAT2 cause lipodystrophy, CGI-58 mutations cause Chanarin-Dorfman syndrome, and altered activity is linked to cancer and metabolic disorders.
How is LPAAT activity measured?
Common methods include radioenzymatic assays, LC-MS lipidomics, and fluorescent substrate assays.
What is the role of LPAAT in seed oil accumulation?
Plant LPAATs enhance seed oil content by converting LPA to PA, a precursor for triacylglycerols, as shown in Brassica napus and Perilla frutescens.
Can CRISPR be used to study LPAAT function?
Yes, CRISPR knockout, knock-in, and overexpression models are powerful tools to dissect LPAAT roles in lipid metabolism and disease.
What is the substrate specificity of LPAAT?
LPAAT enzymes vary in specificity; some are promiscuous (e.g., Thermus thermophilus) while others prefer unsaturated acyl-CoAs.
Is LPAAT a drug target?
Yes, LPAAT enzymes are being explored as targets for cancer, metabolic diseases, and bacterial infections.
What is the difference between LPAAT and AGPAT?
AGPAT (1-acylglycerol-3-phosphate O-acyltransferase) is another name for LPAAT; both catalyze the same reaction.
How does LPAAT relate to phosphatidic acid signaling?
By producing phosphatidic acid, LPAAT activity influences signaling pathways involved in cell growth and survival.
Conclusion
Lysophosphatidic acid acyltransferase activity (GO:0042171) is a fundamental enzymatic function with far-reaching implications in lipid metabolism, human disease, and plant oil production. The conserved mechanism and diverse isoforms offer rich opportunities for basic and applied research. Leveraging CRISPR technologies and advanced lipidomics, researchers can uncover new roles for LPAATs and develop targeted interventions.
References
- 1. Ogawa T et al.. 2020. Lysophosphatidic acid acyltransferase from the thermophilic bacterium Thermus thermophilus HB8 displays substrate promiscuity.. Biosci Biotechnol Biochem 84(9):1831-1838 PMID: 32456605
- 2. Zhang K et al.. 2022. Lysophosphatidic acid acyltransferase 2 and 5 commonly, but differently, promote seed oil accumulation in Brassica napus.. Biotechnol Biofuels Bioprod 15(1):83 PMID: 35962411
- 3. Zhang X et al.. 2019. Identification of an intrinsic lysophosphatidic acid acyltransferase activity in the lipolytic inhibitor G(0)/G(1) switch gene 2 (G0S2).. FASEB J 33(5):6655-6666 PMID: 30802154
- 4. Zhukovsky MA et al.. 2019. The Structure and Function of Acylglycerophosphate Acyltransferase 4/ Lysophosphatidic Acid Acyltransferase Delta (AGPAT4/LPAATδ).. Front Cell Dev Biol 7:147 PMID: 31428612
- 5. Zhou Y et al.. 2022. [Cloning and functional characterization of a lysophosphatidic acid acyltransferase gene from Perilla frutescens].. Sheng Wu Gong Cheng Xue Bao 38(8):3014-3028 PMID: 36002428
- 6. Cullinane M et al.. 2005. Identification of two lysophosphatidic acid acyltransferase genes with overlapping function in Pseudomonas fluorescens.. Microbiology (Reading) 151(Pt 9):3071-3080 PMID: 16151217
- 7. Montero-Moran G et al.. 2010. CGI-58/ABHD5 is a coenzyme A-dependent lysophosphatidic acid acyltransferase.. J Lipid Res 51(4):709-19 PMID: 19801371
- 8. Bourgis F et al.. 1999. A plastidial lysophosphatidic acid acyltransferase from oilseed rape.. Plant Physiol 120(3):913-22 PMID: 10398728
