PLA2G10 Human
Description
Role in Inflammation and Immunity
PLA2G10 drives allergic and inflammatory responses by:
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Generating lipid mediators (e.g., leukotrienes) that recruit eosinophils and T cells .
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Enhancing IL-33 release from epithelial cells, activating type-2 innate lymphoid cells (ILC2s) and macrophages .
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Polarizing macrophages toward an M2 phenotype, promoting tissue remodeling in asthma .
In murine models, Pla2g10 deletion reduced airway hyperresponsiveness by 60% and eosinophil infiltration by 75% .
Cancer Immunomodulation
PLA2G10 is upregulated in lung, pancreatic, and prostate cancers, correlating with poor T cell infiltration . Mechanisms include:
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Hydrolyzing phospholipids to disrupt chemokine gradients, impairing T cell migration .
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Reducing CD8+ T cell presence in tumors by 40–50%, diminishing anti-PD-1 therapy efficacy .
Disease Associations
Inhibitors and Antibodies
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Monoclonal antibodies targeting PLA2G10 restored T cell infiltration in murine tumors by 70%, enhancing anti-PD-1 response .
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Small-molecule sPLA2 inhibitors (e.g., varespladib) are under investigation for inflammatory diseases .
Diagnostic Potential
PLA2G10 levels in bronchoalveolar lavage fluid (BALF) correlate with asthma severity (r = 0.62, p < 0.01) .
Research Applications
Product Specs
MRGSHHHHHH GMASHMGILE LAGTVGCVGP RTPIAYMKYG CFCGLGGHGQ PRDAIDWCCH GHDCCYTRAE EAGCSPKTER YSWQCVNQSV LCGPAENKCQ ELLCKCDQEI ANCLAQTEYN LKYLFYPQFL CEPDSPKCD
Q&A
What is PLA2G10 and what are its primary functions in human biology?
PLA2G10 is a secreted phospholipase that catalyzes the hydrolysis of glycerophospholipids at the sn-2 position, releasing fatty acids (particularly arachidonic acid) and lysophospholipids. Its primary functions include:
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Contribution to cellular response to leukemia inhibitory factor
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Modulation of inflammatory responses through negative regulation of cytokine production
Methodological approach: To study PLA2G10's basic functions, researchers typically employ enzymatic activity assays using fluorescent or radiolabeled substrates, coupled with mass spectrometry-based lipidomics to analyze changes in phospholipid composition and fatty acid release patterns.
How is PLA2G10 expression regulated in different tissue contexts?
PLA2G10 expression exhibits tissue-specific patterns and is dysregulated in various cancer types:
Normal tissues:
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Tightly regulated expression with relatively low levels in most cell types
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Modulation by inflammatory signals and cytokines
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Regulation in response to lipid metabolism needs
Cancerous tissues:
Methodological approach: Expression analysis typically combines quantitative PCR, Western blotting, and immunohistochemistry. Single-cell RNA sequencing can identify specific cell populations expressing PLA2G10 within heterogeneous tumor tissues.
What are the structural characteristics of human PLA2G10 relevant to its function?
Human PLA2G10's structure includes several key features that define its enzymatic function:
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A calcium-binding domain essential for catalytic activity
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Conserved disulfide bonds maintaining tertiary structure
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A catalytic site with a histidine residue critical for phospholipid hydrolysis
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A hydrophobic channel accommodating fatty acid chains
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A C2-like domain involved in calcium-dependent membrane binding
Methodological approach: Structural characterization typically employs X-ray crystallography complemented by site-directed mutagenesis to analyze structure-function relationships. Molecular dynamics simulations help understand protein flexibility and substrate interactions.
How does PLA2G10 contribute to phospholipid metabolism?
PLA2G10 plays a central role in phospholipid metabolism through:
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Catalyzing the hydrolysis of the sn-2 ester bond in glycerophospholipids
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Releasing free fatty acids (particularly arachidonic acid) and lysophospholipids
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Contributing to membrane remodeling and phospholipid turnover
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Participating in low-density lipoprotein particle remodeling
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Influencing lipid composition in cellular membranes
Methodological approach: Researchers employ mass spectrometry-based lipidomics to analyze phospholipid composition changes, combined with enzymatic activity assays and cellular phospholipid turnover studies using isotope labeling.
What experimental methods are used to measure PLA2G10 enzymatic activity?
Multiple complementary approaches are employed to measure PLA2G10 activity:
| Method | Description | Advantages | Limitations |
|---|---|---|---|
| Fluorogenic substrate assays | Uses substrates that generate fluorescent products upon hydrolysis | High sensitivity, real-time monitoring | May lack specificity for PLA2G10 |
| Radiometric assays | Measures release of radiolabeled fatty acids from phospholipid substrates | High sensitivity, quantitative | Requires special handling for radioactive materials |
| HPLC-MS analysis | Detects and quantifies specific fatty acids released | High specificity, comprehensive analysis | Requires specialized equipment, time-consuming |
| Colorimetric assays | Uses pH indicators or chromogenic substrates | Simple to perform, economical | Lower sensitivity, potential interference |
Methodological approach: For most accurate results, researchers should combine multiple methods and include appropriate controls with selective inhibitors to determine the contribution of PLA2G10 to total phospholipase activity.
What mechanisms explain PLA2G10's role in T cell exclusion from the tumor microenvironment?
Recent research has identified PLA2G10 as a key factor in T cell exclusion from tumors. The mechanisms include:
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Impairment of chemokine-mediated T cell migration through phospholipid hydrolysis
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Creation of a lipid mediator profile in the tumor microenvironment that inhibits T cell infiltration
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Potential disruption of T cell receptor signaling required for tumor infiltration
A genome-wide screen of >1000 soluble human proteins identified PLA2G10 as a candidate driver of T cell exclusion in chemokine-mediated T cell migration . Experimental validation demonstrated that overexpression of PLA2G10 in immunologically "hot" murine tumor lines prevented T cell infiltration and resulted in resistance to anti-PD-1 therapy .
Methodological approach: Researchers investigating this mechanism employ transwell migration assays with conditioned media from PLA2G10-expressing tumor cells, in vivo tumor models with genetic manipulation of PLA2G10 expression, and multiphoton intravital microscopy to visualize T cell migration in real-time.
How does PLA2G10 affect response to immune checkpoint inhibitors?
PLA2G10 expression has significant implications for immunotherapy efficacy:
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Tumors with high PLA2G10 expression show reduced responsiveness to anti-PD-1 therapy
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The mechanism involves impaired T cell infiltration into the tumor microenvironment
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PLA2G10 creates an immunosuppressive microenvironment that cannot be fully overcome by checkpoint blockade alone
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The enzymatic activity of PLA2G10 is critical for this immune evasion phenomenon
Methodological approach: Studies typically combine preclinical models with modulated PLA2G10 expression treated with immune checkpoint inhibitors, flow cytometric analysis of tumor-infiltrating lymphocytes, and patient tumor sample analyses correlating PLA2G10 expression with immunotherapy response.
What is the relationship between PLA2G10 expression and cell cycle regulation in cancer cells?
PLA2G10 significantly influences cell cycle progression in cancer cells:
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In soft tissue leiomyosarcoma (STLMS), PLA2G10 facilitates cell cycle progression by elevating cyclin E1/CDK2 expression
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The enzyme has been identified as specifically associated with relapse in STLMS
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PLA2G10-derived lipid mediators can activate signaling pathways that promote proliferation
Methodological approach: Researchers employ flow cytometry with DNA dyes to assess cell cycle distribution, BrdU incorporation assays to measure DNA synthesis, and Western blotting for cell cycle proteins (cyclins, CDKs, CDK inhibitors) in cells with modulated PLA2G10 expression.
What are the most effective approaches for PLA2G10 inhibition in experimental models?
Multiple strategies can effectively inhibit PLA2G10 in research settings:
| Approach | Examples | Advantages | Considerations |
|---|---|---|---|
| Pharmacological inhibition | Small molecule inhibitors, substrate analogs | Rapid action, dose-dependent effects | Potential off-target effects |
| Genetic approaches | siRNA, shRNA, CRISPR-Cas9 | High specificity, complete elimination possible | Longer implementation time, delivery challenges |
| Biological inhibitors | Neutralizing antibodies, recombinant binding proteins | High specificity, effective for secreted PLA2G10 | May not reach intracellular enzyme |
| Expression modulation | Epigenetic modulators, transcription factor inhibitors | Targets endogenous regulation mechanisms | Indirect effects, potential broad impact |
Methodological approach: Selection of inhibition method should consider specificity requirements, duration of inhibition needed, and experimental system (in vitro vs. in vivo), with validation through enzymatic activity assays and downstream functional assessments.
How does PLA2G10 influence the lipid mediator profile in the tumor microenvironment?
PLA2G10 enzymatic activity generates various lipid mediators that shape the tumor microenvironment:
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Arachidonic acid release serves as a precursor for prostaglandins, leukotrienes, and other eicosanoids
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Lysophospholipids produced act as signaling molecules affecting various cellular functions
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The specific profile of fatty acids released depends on the phospholipid composition of cellular membranes
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These lipid mediators collectively contribute to T cell exclusion and resistance to immunotherapy
Methodological approach: Lipidomic analysis using liquid chromatography-mass spectrometry (LC-MS/MS) is the gold standard for comprehensive lipid mediator profiling, complemented by spatial mapping techniques and functional validation using recombinant lipid mediators.
What experimental models are most appropriate for studying PLA2G10 function in cancer?
Several experimental models have proven valuable for investigating PLA2G10 in cancer:
| Model Type | Examples | Best Applications | Limitations |
|---|---|---|---|
| In vitro cell models | Cancer cell lines with engineered PLA2G10 expression | Mechanism studies, high-throughput screening | Limited microenvironmental complexity |
| 3D culture systems | Organoids, spheroids, co-cultures | Cell-cell interactions, spatial organization | Still lacks full in vivo complexity |
| Murine tumor models | Syngeneic models with PLA2G10 manipulation | Immune interactions, therapy response | Species differences in lipid metabolism |
| Patient-derived models | PDX, primary tumor cultures | Human relevance, personalized approaches | Often lacks intact immune components |
Research has successfully employed PLA2G10 overexpression in murine tumor lines to study its impact on T cell infiltration and immunotherapy response . For soft tissue leiomyosarcoma, cell line models have been developed to study PLA2G10's role in cell cycle progression .
Methodological approach: The choice of model should be guided by the specific research question, with consideration of immune system interactions, relevance to human cancer biology, and technical feasibility.
How can researchers distinguish PLA2G10 activity from other phospholipases?
Differentiating PLA2G10 activity from other phospholipases requires multiple complementary approaches:
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Substrate preference profiling using structurally diverse phospholipids
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Analysis of positional specificity (sn-1 vs. sn-2 position)
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pH and calcium dependence studies (PLA2G10 has specific requirements)
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Product profile characterization through mass spectrometry
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Selective genetic manipulation (knockdown, knockout, overexpression)
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Use of selective inhibitors with known specificity profiles
Methodological approach: Researchers typically combine enzymatic assays under various conditions with genetic manipulation approaches and product characterization to confidently attribute observed effects to PLA2G10 rather than other phospholipases.
What are the potential downstream targets of PLA2G10 in tumor progression?
PLA2G10 influences tumor progression through multiple downstream targets:
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Cell cycle regulators: Upregulation of cyclin E1/CDK2 expression
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Inflammatory mediators: Modulation of prostaglandins and leukotrienes
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Transcription factors: Negative regulation of DNA-binding transcription factor activity
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Immune regulators: Alteration of chemokine gradients and cytokine production
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Signaling pathways: Impact on MAPK, PI3K/AKT, and nuclear receptor signaling
Methodological approach: Identification of downstream targets employs phosphoproteomics, transcriptomic analysis, targeted inhibitor studies, and genetic rescue experiments to confirm specificity of observed effects.
How do genetic variants of PLA2G10 affect its enzymatic activity and disease associations?
Genetic variation in PLA2G10 can impact its function and disease associations:
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Coding variants may alter catalytic efficiency, substrate specificity, or protein stability
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Promoter polymorphisms can affect expression levels and tissue distribution
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Splice variants may generate protein isoforms with distinct activities
Methodological approach: Studies combine genotyping and sequencing with expression quantitative trait locus (eQTL) analysis, recombinant protein production of variant forms for enzymatic characterization, and introduction of specific variants into cell or animal models for functional assessment.
What technologies are emerging for temporal and spatial control of PLA2G10 activity in research?
Advanced technologies for precise control of PLA2G10 activity include:
| Technology | Approach | Research Applications |
|---|---|---|
| Optogenetics | Light-controlled gene expression or protein activation | Spatiotemporal control of PLA2G10 expression in complex tissues |
| Chemogenetics | Chemical-dependent activation of modified proteins | Dose-dependent and reversible control of PLA2G10 activity |
| CRISPR interference | Inducible CRISPR systems for temporal gene regulation | Controlled knockdown with precise timing |
| Nanoparticle delivery | Targeted delivery of inhibitors or siRNA | Tissue-specific inhibition in vivo |
| Photocaged inhibitors | Light-activated release of enzyme inhibitors | Precise spatial control of inhibition |
Methodological approach: Selection of technology depends on the specific research question, with considerations for temporal precision, spatial resolution, reversibility requirements, and compatibility with the experimental system.
Product Science Overview
Secreted Phospholipase A2-X (sPLA2-X) is a member of the secreted phospholipase A2 (sPLA2) family, which consists of low molecular weight, calcium-dependent enzymes. These enzymes are known for their ability to hydrolyze the sn-2 position of glycerophospholipids, resulting in the production of free fatty acids and lysophospholipids .
sPLA2-X, like other members of the sPLA2 family, contains a conserved His-Asp catalytic dyad essential for its enzymatic activity . This enzyme is unique due to its specific localization and distinct enzymatic properties, which suggest specialized biological roles . sPLA2-X is involved in the metabolism of phospholipids, playing a crucial role in the production of bioactive lipid mediators such as prostaglandins and leukotrienes .
Historically, sPLA2 enzymes have been implicated in various biological processes, including inflammation and atherosclerosis . sPLA2-X, in particular, has been shown to promote inflammation by catalyzing the first step of the arachidonic acid pathway, leading to the formation of inflammatory and thrombogenic molecules . Recent studies using transgenic and knockout mouse models have revealed the distinct roles of sPLA2-X in various biological events, including metabolic disorders such as obesity, hepatic steatosis, diabetes, insulin resistance, and adipose tissue inflammation .
Human recombinant sPLA2-X is produced using advanced biotechnological methods to ensure high purity and activity. This recombinant enzyme is widely used in research to study its biological functions and potential therapeutic applications. By understanding the mechanisms of sPLA2-X, researchers aim to develop novel treatments for inflammatory diseases and metabolic disorders .
