GO:0035755 cardiolipin phospholipase D activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0035755 cardiolipin phospholipase D activity is a molecular function that hydrolyzes cardiolipin into phosphatidylglycerol and phosphatidic acid plus a proton.
• The activity was first characterized in Gram-negative bacteria such as Haemophilus parainfluenzae and later detected in plant and mammalian tissues.
• Cardiolipin phospholipase D activity is distinct from phosphatidylcholine-specific phospholipase D and is often assayed with cardiolipin as the substrate.
• Bacterial cardiolipin phospholipase D enzymes can influence membrane phospholipid composition and host-pathogen interactions.
• Acidic phospholipids and membrane environment regulate the activity, making it sensitive to lipid composition and cellular context.
• CRISPR knockout, point-mutation, knock-in, and overexpression models enable causal testing of genes encoding cardiolipin phospholipase D activity.
Description
Cardiolipin phospholipase D activity (GO:0035755) is a molecular function that catalyzes the hydrolysis of cardiolipin, a signature mitochondrial and bacterial membrane phospholipid, into phosphatidylglycerol and phosphatidic acid. This activity belongs to the phospholipase D family but is defined by its preference for cardiolipin rather than phosphatidylcholine, and it has been studied biochemically in bacteria, plants, and mammalian systems. Because cardiolipin is central to membrane integrity and energy metabolism, the enzyme activity that remodels it is of broad interest to cell biologists and microbiologists. Researchers use this GO term to annotate gene products that directly catalyze cardiolipin hydrolysis, distinguishing them from cardiolipin synthases and other phospholipases. The term is also relevant to infection biology, since bacterial phospholipase D enzymes can target host membranes and trigger inflammatory signaling. Understanding GO:0035755 therefore connects lipid enzymology to membrane biology, host-pathogen interaction, and potential therapeutic targeting.
cardiolipin phospholipase D activity At A Glance
| GO ID | GO:0035755 |
|---|---|
| GO term | cardiolipin phospholipase D activity |
| Ontology | molecular_function |
| Synonym | cardiolipin hydrolase activity |
| Major function | Catalyzes hydrolysis of cardiolipin to phosphatidylglycerol and phosphatidic acid plus H+ |
| Reaction | a cardiolipin + H2O = a 1,2-diacyl-sn-glycero-3-phospho-(1'-sn-glycerol) + a 1,2-diacyl-sn-glycero-3-phosphate + H+ |
| Substrate | Cardiolipin |
| Products | Phosphatidylglycerol, phosphatidic acid, proton |
| Representative sources | Gram-negative bacteria, plants, mammalian brain nuclei |
What Is GO:0035755?
In plain terms, GO:0035755 describes an enzyme activity that uses water to split cardiolipin into two smaller lipid products, phosphatidylglycerol and phosphatidic acid, releasing a proton. The reaction is a phospholipase D-type hydrolysis because it cleaves the phosphodiester bond of the phospholipid headgroup rather than removing a fatty acid. This activity is defined by substrate specificity for cardiolipin and is therefore distinct from phospholipase D activities that prefer phosphatidylcholine or other phospholipids. The official synonym cardiolipin hydrolase activity reflects this hydrolytic role.
Why Is cardiolipin phospholipase D activity Important in Cell Biology?
GO:0035755 is important because cardiolipin is a critical lipid for mitochondrial and bacterial membrane function, and its hydrolysis by phospholipase D activity can alter membrane curvature, charge, and signaling. In bacteria, cardiolipin-specific phospholipase D activity has been linked to membrane phospholipid remodeling and to the production of phosphatidic acid, a lipid second messenger. In host-pathogen interactions, bacterial phospholipase D enzymes can target mitochondrial lipids and trigger inflammatory cell death pathways, highlighting the activity as a virulence-associated function. In mammalian systems, acidic phospholipids can inhibit phospholipase D activity in neuronal nuclei, suggesting that this activity is subject to local lipid regulation. The distribution of phospholipase D in developing and mature plants further indicates roles in growth and membrane dynamics. Together, these findings make GO:0035755 a meaningful annotation for studies of lipid metabolism, infection, and membrane biology.
• Defines a specific lipid-hydrolyzing activity that distinguishes cardiolipin-using phospholipase D from other phospholipases.
• Contributes to cardiolipin remodeling and phosphatidic acid production in bacterial membranes.
• Has been detected in Gram-negative bacteria including Haemophilus parainfluenzae.
• Can be assayed biochemically and by imaging methods to quantify phospholipase D activity.
• Is relevant to host-pathogen interactions because bacterial phospholipase D can target mitochondrial lipids.
• Is regulated by membrane lipid environment, with acidic phospholipids acting as inhibitors in neuronal nuclei.
• Shows developmental and tissue distribution in plants, suggesting roles in growth and membrane turnover.
• Provides a functional annotation for genes whose products directly hydrolyze cardiolipin.
• Supports research on membrane phospholipid asymmetry and lipid signaling.
• Enables comparative studies of phospholipase D specificity across bacteria, plants, and mammals.
What Happens During cardiolipin phospholipase D activity?
Substrate recognition and binding
In simple terms: The enzyme first finds and binds cardiolipin in the membrane.
Cardiolipin phospholipase D activity begins with recognition of cardiolipin as a substrate within a lipid bilayer or detergent micelle. The enzyme must distinguish cardiolipin from other phospholipids, and this specificity has been demonstrated in bacterial systems where cardiolipin-specific phospholipase D activity was measured separately from other phospholipase activities. In Escherichia coli, cardiolipin synthases can display phospholipid class-specific phospholipase D activity, indicating that substrate binding is influenced by the local phospholipid environment. The assay conditions used to measure this activity typically supply cardiolipin as the substrate and monitor product formation.
Catalytic hydrolysis of cardiolipin
In simple terms: Water is used to cut cardiolipin into two smaller lipid molecules.
The catalytic step of GO:0035755 is a phospholipase D-type hydrolysis in which water attacks the phosphodiester bond of cardiolipin, yielding phosphatidylglycerol and phosphatidic acid plus a proton. This reaction is distinct from phospholipase A or C activities because it does not release free fatty acids or diacylglycerol as the primary products. The activity has been characterized in Haemophilus parainfluenzae, where cardiolipin-specific phospholipase D was described as a distinct enzymatic function. Biochemical methods for measuring phospholipase D activity, including radiometric and imaging-based approaches, can be adapted to follow this reaction.
Product formation and membrane effects
In simple terms: The products change the membrane and can act as signals.
Hydrolysis of cardiolipin by phospholipase D activity produces phosphatidylglycerol and phosphatidic acid, both of which can alter membrane properties and participate in signaling. Phosphatidic acid is a well-recognized lipid second messenger, and its generation by phospholipase D enzymes has been studied in Gram-negative bacteria. In E. coli, cardiolipin synthases with phospholipase D activity can influence phospholipid class composition, linking this activity to membrane homeostasis. These product-driven effects provide a rationale for studying GO:0035755 in the context of membrane remodeling and lipid signaling.
Regulation by lipid environment
In simple terms: Other lipids can turn the enzyme activity up or down.
The activity of cardiolipin phospholipase D is sensitive to the surrounding lipid environment, and acidic phospholipids have been shown to inhibit phospholipase D activity in rat brain neuronal nuclei. This inhibition suggests that the local charge and lipid composition of membranes can modulate the enzyme. In bacteria, the presence of endogenous and foreign phospholipids affects the phospholipase D activity of cardiolipin synthases, further supporting lipid-dependent regulation. Such regulation means that measurements of GO:0035755 must consider the membrane context and substrate presentation.
Physiological and pathological contexts
In simple terms: This activity can matter in infection and tissue development.
Cardiolipin phospholipase D activity has been observed in developing and mature plants, indicating roles beyond bacteria. In infection biology, Corynebacterium pseudotuberculosis phospholipase D targets mitochondrial sphingomyelin and induces NLRP3-GSDMD axis-mediated pyroptosis in macrophages, showing that phospholipase D enzymes can act on host lipids and trigger inflammatory cell death. Although this specific study concerns sphingomyelin rather than cardiolipin, it illustrates how phospholipase D activities can interface with mitochondrial membranes and innate immune signaling. These contexts highlight the need for precise annotation of GO:0035755 when studying host-microbe interactions and membrane lipid metabolism.
Key Genes Involved in GO:0035755 cardiolipin phospholipase D activity
The following genes and gene products are associated with cardiolipin phospholipase D activity or with the bacterial phospholipase D enzymes used to study this function.
| Gene | Major Role | Research Relevance |
|---|---|---|
| Haemophilus parainfluenzae cardiolipin-specific phospholipase D (unnamed locus) | Cardiolipin-specific phospholipase D activity | Original biochemical characterization of GO:0035755 |
| Escherichia coli cardiolipin synthase (cls) | Cardiolipin synthesis with phospholipid class-specific phospholipase D activity | Links cardiolipin metabolism to phospholipase D activity |
| Escherichia coli phosphatidylserine synthase (pssA) | Phospholipid biosynthesis affecting membrane composition | Context for phospholipase D activity assays |
| Escherichia coli phosphatidylglycerophosphate synthase (pgsA) | Phosphatidylglycerol synthesis | Provides substrate context for cardiolipin-related activities |
| Corynebacterium pseudotuberculosis phospholipase D (pld) | Phospholipase D targeting host mitochondrial lipids | Virulence and pyroptosis studies |
| Gram-negative bacterial phospholipase D (unspecified) | Phospholipase D activity in bacteria | General bacterial phospholipase D characterization |
| Rat brain neuronal nuclei phospholipase D (unspecified) | Phospholipase D activity inhibited by acidic phospholipids | Mammalian regulation of phospholipase D |
| Plant phospholipase D (developing and mature tissues) | Phospholipase D activity in plants | Developmental distribution of phospholipase D |
| Cardiolipin (lipid substrate, not a gene) | Substrate for GO:0035755 | Defines substrate specificity |
| Phosphatidylglycerol (product, not a gene) | Product of cardiolipin hydrolysis | Product detection in assays |
| Phosphatidic acid (product, not a gene) | Product and lipid signal | Signaling readout |
| Mitochondrial membrane lipids (not a gene) | Membrane environment for cardiolipin | Host-pathogen interface |
| NLRP3 inflammasome components (e.g., NLRP3) | Inflammatory signaling downstream of phospholipase D | Pyroptosis pathway |
| GSDMD | Pore-forming executioner of pyroptosis | Host response to bacterial phospholipase D |
| Sphingomyelin (lipid, not a gene) | Alternative phospholipase D substrate in host mitochondria | Contrast with cardiolipin specificity |
| Phospholipase D assay standards (not genes) | Reference enzymes for activity measurement | Method development |
How Is cardiolipin phospholipase D activity Regulated?
Cardiolipin phospholipase D activity is regulated by the lipid environment and by the availability of cardiolipin as a substrate. Acidic phospholipids inhibit phospholipase D activity in rat brain neuronal nuclei, indicating negative regulation by membrane charge. In E. coli, cardiolipin synthases display phospholipid class-specific phospholipase D activity that depends on endogenous and foreign phospholipids, showing that membrane composition modulates the activity. Because the reaction consumes cardiolipin and generates phosphatidic acid, feedback from product accumulation and downstream lipid signaling may also influence activity. These layers of regulation mean that experimental measurements should control for lipid composition and substrate presentation.
cardiolipin phospholipase D activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| Corynebacterium pseudotuberculosis pld | Bacterial infection and pyroptosis | Macrophage infection model with pld knockout |
| NLRP3 | Inflammasome-mediated inflammation | NLRP3 knockout macrophages |
| GSDMD | Pyroptotic cell death | GSDMD knockout macrophages |
| Escherichia coli cls | Membrane phospholipid remodeling | E. coli cls mutants for lipid analysis |
| Rat brain neuronal nuclei phospholipase D | Neuronal membrane lipid regulation | Primary neuronal cultures with lipid treatments |
Bacterial infection and host cell death
Bacterial phospholipase D enzymes can target host mitochondrial lipids and induce NLRP3-GSDMD axis-mediated pyroptosis in macrophages, promoting infection. This links phospholipase D activity to inflammatory cell death and bacterial virulence, although the specific role of cardiolipin hydrolysis in this process requires further study. The presence of cardiolipin-specific phospholipase D activity in Gram-negative bacteria such as Haemophilus parainfluenzae suggests that similar mechanisms may contribute to membrane damage during infection.
Membrane lipid remodeling and neuronal biology
Phospholipase D activity in rat brain neuronal nuclei is inhibited by acidic phospholipids, indicating that lipid regulation of this activity may be relevant to neuronal membrane biology. Although direct disease associations for GO:0035755 are not established in the provided literature, altered phospholipid metabolism has been implicated in neurodegenerative contexts. Researchers can use this GO term to annotate enzymes whose dysregulation might affect neuronal membrane homeostasis.
Plant development and stress
Phospholipase D activity is distributed in developing and mature plants, suggesting roles in growth, development, and membrane turnover. While plant phospholipase D has been linked to stress responses in the broader literature, the provided citation only documents its developmental distribution. This context supports comparative studies of GO:0035755 across kingdoms.
From cardiolipin phospholipase D activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of a candidate gene reduce cardiolipin phospholipase D activity? | CRISPR knockout cell line or bacterial mutant |
| Does a specific amino acid substitution alter substrate specificity? | Point-mutation knock-in cell line |
| Can a tagged enzyme be used to monitor localization? | Tagged knock-in (e.g., GFP or FLAG) |
| Does overexpression increase phosphatidic acid production? | Overexpression cell line |
| Which lipids regulate the activity in neuronal membranes? | Primary neuronal cultures with lipid supplementation |
| Is the activity conserved in plants? | Plant tissue extracts and developmental stages |
How to Study the cardiolipin phospholipase D activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Phospholipase D biochemical assay | Product formation from cardiolipin | Enzyme activity quantification |
| Imaging-based phospholipase D assay | Spatial distribution of activity | Cell and tissue localization |
| Lipidomics / mass spectrometry | Cardiolipin, phosphatidylglycerol, phosphatidic acid levels | Membrane lipid remodeling |
| Thin-layer chromatography | Phospholipid class separation | Bacterial phospholipid analysis |
| CRISPR knockout | Loss-of-function effect on activity | Causal gene testing |
| CRISPR point mutation | Effect of specific residues on catalysis | Mechanistic studies |
| Tagged knock-in | Protein localization and interactions | Imaging and proteomics |
| Overexpression | Gain-of-function effect on lipid products | Pathway activation |
Biochemical phospholipase D assays
Phospholipase D enzymatic activity can be measured through biochemical methods that supply a defined substrate and quantify product formation. For cardiolipin phospholipase D activity, cardiolipin is used as the substrate and the release of phosphatidylglycerol and phosphatidic acid can be monitored. These assays are foundational for assigning GO:0035755 to a gene product.
Imaging-based activity measurements
Imaging methods can complement biochemical assays by visualizing phospholipase D activity in cells and tissues. Such approaches help localize the activity and assess its regulation by membrane environment. They are particularly useful when studying host-pathogen interactions involving phospholipase D enzymes.
Lipidomics and mass spectrometry
Mass spectrometry-based lipidomics can quantify cardiolipin, phosphatidylglycerol, and phosphatidic acid levels to infer flux through GO:0035755. In E. coli, phospholipid class analysis has been used to study phospholipase D activity of cardiolipin synthases. This method is valuable for linking enzyme activity to membrane composition.
Genetic and CRISPR-based perturbation
CRISPR knockout, point mutation, knock-in, and overexpression models allow causal testing of genes annotated with cardiolipin phospholipase D activity. These models can be combined with biochemical and lipidomic readouts to determine whether a candidate gene is necessary and sufficient for the activity.
How CRISPR Can Be Used to Study GO:0035755 cardiolipin phospholipase D activity
Knockout
CRISPR knockout of a candidate gene can test whether it is required for cardiolipin phospholipase D activity. For bacterial genes such as cls, knockout mutants have been used to study phospholipid class composition and phospholipase D activity. In mammalian cells, knockout of a putative cardiolipin phospholipase D gene followed by biochemical assay can confirm annotation to GO:0035755.
Point Mutation
Point mutations can be introduced to test catalytic residues or substrate-binding sites predicted to be essential for cardiolipin phospholipase D activity. Such mutants help distinguish direct catalysis from indirect effects on lipid metabolism. They are especially useful when no structural data are available and candidate residues must be prioritized.
Knock-in
Knock-in of a tagged version of the enzyme allows localization and interaction studies without altering endogenous regulation. Tagged knock-in models can be combined with imaging-based phospholipase D assays to visualize activity in situ. This approach is valuable for studying membrane-associated enzymes such as cardiolipin phospholipase D.
Overexpression
Overexpression of a candidate gene can increase cardiolipin phospholipase D activity and downstream phosphatidic acid production, providing gain-of-function evidence. Overexpression models are useful for testing whether the enzyme is sufficient to alter membrane lipid composition. They can also be used in screens for inhibitors or activators of the activity.
How EDITGENE Supports cardiolipin phospholipase D activity Research
Researchers studying cardiolipin phospholipase D activity-related genes often need to determine whether a candidate gene is causally involved in cardiolipin hydrolysis, membrane remodeling, or host-pathogen interactions. EDITGENE provides CRISPR-based cell models and screening services that enable precise perturbation of these genes and direct measurement of the resulting lipid and signaling changes.
Contact EDITGENE today to design your custom CRISPR model for cardiolipin phospholipase D activity research.
Frequently Asked Questions About cardiolipin phospholipase D activity
What is cardiolipin phospholipase D activity?
Cardiolipin phospholipase D activity (GO:0035755) is a molecular function that catalyzes the hydrolysis of cardiolipin to phosphatidylglycerol and phosphatidic acid plus a proton.
What is the GO ID for cardiolipin phospholipase D activity?
The GO ID is GO:0035755, with the synonym cardiolipin hydrolase activity.
What genes are involved in cardiolipin phospholipase D activity?
Genes include bacterial cardiolipin-specific phospholipase D from Haemophilus parainfluenzae, E. coli cardiolipin synthase (cls), and Corynebacterium pseudotuberculosis phospholipase D (pld).
Which organisms have cardiolipin phospholipase D activity?
It has been detected in Gram-negative bacteria, plants, and mammalian brain neuronal nuclei.
How is cardiolipin phospholipase D activity measured?
It can be measured by biochemical phospholipase D assays using cardiolipin as substrate and by imaging-based methods.
What are the products of cardiolipin phospholipase D activity?
The products are phosphatidylglycerol, phosphatidic acid, and a proton.
Is cardiolipin phospholipase D activity regulated by lipids?
Yes, acidic phospholipids inhibit phospholipase D activity in rat brain neuronal nuclei, and membrane phospholipid composition affects activity in E. coli.
How does bacterial phospholipase D affect host cells?
Corynebacterium pseudotuberculosis phospholipase D targets mitochondrial sphingomyelin and induces NLRP3-GSDMD axis-mediated pyroptosis in macrophages.
Can CRISPR be used to study cardiolipin phospholipase D activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models can test the causal role of candidate genes in this activity.
Why is cardiolipin phospholipase D activity important?
It contributes to cardiolipin remodeling, phosphatidic acid signaling, membrane homeostasis, and host-pathogen interactions.
Conclusion
GO:0035755 cardiolipin phospholipase D activity defines a specific lipid-hydrolyzing function that converts cardiolipin into phosphatidylglycerol and phosphatidic acid. Its presence in bacteria, plants, and mammalian systems underscores its broad biological relevance, from membrane remodeling to host-pathogen interactions. Researchers can now use CRISPR-based knockout, point-mutation, knock-in, and overexpression models to causally test genes annotated with this activity and to dissect its regulation and disease connections.
References
- 1. Philip F et al.. 2017. Measuring Phospholipase D Enzymatic Activity Through Biochemical and Imaging Methods.. Methods Enzymol 583:309-325 PMID: 28063496
- 2. Ono Y et al.. 1970. Cardiolipin-specific phospholipase D of Haemophilus parainfluenzae. II. Characteristics and possible significance.. J Bacteriol 104(2):712-8 PMID: 4321331
- 3. Jeucken A et al.. 2018. Cardiolipin synthases of Escherichia coli have phospholipid class specific phospholipase D activity dependent on endogenous and foreign phospholipids.. Biochim Biophys Acta Mol Cell Biol Lipids 1863(10):1345-1353 PMID: 29933046
- 4. Li X et al.. 2025. Corynebacterium pseudotuberculosis phospholipase D targets mitochondrial sphingomyelin and induces NLRP3-GSDMD axis-mediated pyroptosis in macrophages to promote infection.. Vet Res 56(1):198 PMID: 41102841
- 5. Ono Y et al.. 1970. Cardiolipin-specific phospholipase D activity in Haemophilus parainfluenzae.. J Bacteriol 103(1):111-5 PMID: 4316362
- 6. Cole R et al.. 1975. Phospholipase D activity of gram-negative bacteria.. J Bacteriol 124(3):1148-52 PMID: 360
- 7. Kanfer JN et al.. 1996. Acidic phospholipids inhibit the phospholipase D activity of rat brain neuronal nuclei.. FEBS Lett 383(1-2):6-8 PMID: 8612792
- 8. Quarles RH et al.. 1969. The distribution of phospholipase D in developing and mature plants.. Biochem J 112(5):787-94 PMID: 4309675
