Recombinant Human 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase epsilon-1 (PLCE1), partial
Description
Molecular Structure and Functional Domains
PLCE1 contains distinct catalytic regions:
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Phospholipase C (PLC) domain: Hydrolyzes PIP2 into IP3 and DAG, critical for intracellular signaling .
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RasGEF domain: Activates Ras GTPases, influencing cell growth and survival .
The recombinant form often excludes non-essential regions to enhance stability or solubility, though specific truncation sites vary by manufacturer .
Expression Systems and Availability
Recombinant PLCE1 is produced in diverse hosts, each offering distinct advantages:
Commercial suppliers like Cusabio and Abbexa provide partial PLCE1 variants with sequence IDs such as CSB-YP868386HU (human) or CSB-EP858169RA (rat) .
Functional Regulation and Post-Translational Modifications
PLCE1 activity is tightly regulated by phosphorylation:
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TAK1-mediated phosphorylation: TAK1 kinase phosphorylates PLCE1 at Serine 1060 (S1060), reducing its PLC activity and downstream IP3/DAG production. This mechanism suppresses PKC/GSK-3β/β-catenin signaling, inhibiting cell migration in esophageal squamous cell carcinoma (ESCC) .
Key Findings on TAK1-PLCE1 Interaction:
Nephrotic Syndrome and PLCE1 Mutations
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Isolated diffuse mesangial sclerosis (IDMS): Truncating mutations in PLCE1 are the most common genetic cause (28.6% of cases) .
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Therapeutic response: Patients with PLCE1 mutations may benefit from cyclosporine therapy, though larger studies are needed .
ELISA-Based Detection
For quantifying PLCE1 levels, Abbexa offers an ELISA kit with the following specifications:
| Parameter | Value |
|---|---|
| Test range | 0.156–10 ng/mL |
| UniProt ID | Q9P212 (PLCE1_HUMAN) |
| Sample types | Tissue lysates, cell cultures, biological fluids |
| Limitations | Not optimized for recombinant proteins |
Research Challenges and Future Directions
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Structural limitations: Partial PLCE1 lacks full-length functionality, complicating studies of RasGEF activity .
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Phosphorylation-specific tools: Limited availability of antibodies for phosphorylated PLCE1 (e.g., p-S1060) hinders mechanistic studies .
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Therapeutic targeting: Inhibiting PLCE1’s PLC activity or enhancing TAK1-mediated repression may offer novel approaches for cancers or fibrotic diseases .
Product Specs
Note: While we preferentially ship the format currently in stock, we are happy to accommodate any special requirements you may have for the format. Please specify your needs when placing the order, and we will prepare the product accordingly.
Note: All our proteins are shipped with standard blue ice packs. If you require dry ice shipment, please inform us in advance. Additional fees will apply.
Generally, the shelf life of liquid forms is 6 months at -20°C/-80°C. Lyophilized forms have a shelf life of 12 months at -20°C/-80°C.
Target Background
- PLCE1 polymorphism has been linked to Gastric and esophageal cancer. PMID: 30202044
- A study found that rs2274223 A>G in PLCE1 was associated with lower PLCE1 gene expression and increased survival in patients with gastric cancer (GC). These findings suggest PLCE1 rs2274223 as a potential prognostic biomarker for GC survival. PMID: 29983348
- The functional characterization of a four-nucleotide insertion/deletion variation (rs71031566 C----/CATTT) in PLCE1, associated with the risk of developing esophageal squamous cell carcinoma, has been reported. PMID: 29106514
- Knockdown of PLCe expression inhibits proliferation of Prostate cancer cells via the PTEN/AKT signaling pathway. PMID: 29330357
- Multiple bioinformatic analyses provide a systematic understanding of PLCE1's roles in esophageal squamous cell carcinoma. PMID: 28849204
- These results indicate that PLCE1 knockdown may play a critical role in controlling esophageal squamous cell carcinoma. PMID: 29238898
- Our findings demonstrate that PLCE1 haplotypes (including rs2274223 and rs3765524) and expression, combined with serum AFP level, may predict the postoperative outcome of HBV-related HCC patients. PMID: 28418898
- High expression of both PLCE1 and PRKCA is significantly associated with poor outcomes in patients with esophageal cancers. PMID: 28402280
- rs10882379 and rs829232 SNPs in the PLCE1 gene may contribute to the susceptibility to esophageal squamous cell carcinoma (ESCC) in the Chinese Han population. Gene-gene and gene-environment interactions also play a crucial role in ESCC progression. PMID: 27061010
- No association between PLCE1 genotype and early viremia level was observed in dengue patients. PMID: 28599625
- PLCE1 single-nucleotide polymorphism is associated with migraine. PMID: 27322543
- PLCE1 expression was detected in invasive carcinoma samples but not in carcinoma in situ samples. Snail expression in human ESCC samples significantly correlated with the PLCE1 protein level, and the expression of Snail target genes in human esophageal cancer samples also correlated with PLCE1 abundance. PMID: 28147304
- These results highlight a novel role of PLCepsilon in maintaining endothelial barrier function through its CDC25 GEF domain and lipase activity, leading to the up-regulation of Rap1 activity. PMID: 27612188
- High PLC epsilon expression is associated with breast cancer. PMID: 28112359
- The mRNA expressions of PPP3CB and MEF2C were significantly up-regulated, while CAMK1 and PPP3R1 were significantly down-regulated in mitral regurgitation (MR) patients compared to normal subjects. Additionally, MR patients had significantly increased mRNA levels of PPP3CB, MEF2C, and PLCE1 compared to aortic valve disease patients. PMID: 27907007
- Two novel putatively deleterious PLCE1 variants were identified in children with steroid-resistant nephrotic syndrome. PMID: 28476686
- PLC-epsilon is a novel regulator of endothelial cell inflammation and vascular endothelial permeability. PMID: 27371732
- Taken together, our data highlight the pivotal role of miR-1976 in the progression of NSCLC. Therefore, miR-1976 may serve as a potential prognostic marker and hold therapeutic relevance for NSCLC progression intervention. PMID: 27063799
- This study investigates the impact of a single nucleotide transition at SNPs associated with DSS on differential protein binding patterns with PLCE1 and MICB genes, and on protein structure modification, highlighting their potential role in the pathogenesis of dengue shock syndrome. PMID: 27038471
- The study shows that PLCE1 Rs2274223 variation was significantly associated with colorectal cancer risk, while rs3765524 was not. PMID: 27383248
- MicroRNA miR-145 was identified as a potent repressor of phospholipase C epsilon 1 (PLCE1) expression by directly targeting the 3' untranslated regions (3'UTR) of PLCE1. PMID: 26657507
- Data suggest the involvement of the phospholipase C epsilon-Protein kinase D-PEA15 protein-ribosomal S6 kinase-IkappaB-NF-kappa B pathway in facilitating inflammation and inflammation-associated carcinogenesis in the colon. PMID: 27053111
- Genetic Variants of PLCE1 Gene are not Associated with Colorectal Cancer. PMID: 26320491
- In conclusion, our results suggest that miR-328 suppresses the survival of EC cells by regulating PLCE1 expression, which might be a potential therapeutic method for EC. PMID: 26773497
- Our results indicated that STAT3 phosphorylation is involved in PLCe-mediated inflammatory cytokine release. PMID: 26156799
- A significantly increased stomach cancer risk was associated with PLCE1 SNP rs2274223 in a Han Chinese population. PMID: 25658482
- Six SNP loci were studied: (rs2279115 of BCL2 gene, rs804270 of NEIL2 gene, rs909253 of LTA gene, rs2294008 of PSCA gene, rs3765524 and rs10509670 of PLCE1 gene) to evaluate gastric cancer risk using magnetic nanoparticles and universal tagged arrays. PMID: 26554163
- This meta-analysis demonstrated that PLCE1 rs2274223 A > G polymorphism may be associated with an increased susceptibility to cancer, particularly for ESCC. PMID: 25614244
- PLCE1 rs2274223 polymorphism is associated with esophageal cancer. PMID: 25854357
- Variation in PLCE1 may be associated with GC risk in the Kashmir Valley. PMID: 25434319
- The meta-analysis suggested that upregulated PLCE1 is significantly associated with an increased risk of tumor progression in esophageal squamous cell carcinoma and gastric cardia adenocarcinomas. PMID: 25520085
- PLCE1 rs2274223 polymorphism may serve as a potential biomarker for digestive tract cancer susceptibility, particularly for esophageal squamous cell carcinoma and gastric cardia adenocarcinomas in the Chinese population. PMID: 24874112
- PLCE1 rs2274223 polymorphism is associated with esophageal cancer. PMID: 25422186
- The rs2274223 polymorphism in phospholipase C epsilon 1 was associated with an increased risk of esophageal squamous cell carcinoma in the Korean population. PMID: 24152165
- Dysregulated PLCE1 mRNA expression was observed for both esophageal squamous cell carcinoma and gastric cancer. PMID: 24867265
- Genetic variations in PLCE1 modulate the risk of esophageal cancer in the high-risk Kashmiri population. PMID: 24935391
- A new pathway for TRPC6 activation by Phospholipase C epsilon was identified. PMID: 25521631
- PLCE1 rs7922612CT + TT is associated with the susceptibility to esophageal cancer. PMID: 25139097
- Findings demonstrate that the presence of the G allele at rs2274223 of the PLCE1 gene may contribute to susceptibility to GC, especially cardia GC. PMID: 24985593
- PLCE1 gene silencing by RNAi resulted in decreased expression of cyclin D1. PMID: 25041015
- Meta-analysis suggested that PLCE1 rs2274223 might act as a cancer risk factor among all subjects, particularly in the Chinese population and upper aerodigestive tract cancer. PMID: 24595086
- Data show that phospholipase C epsilon 1 (PLCE1) and liver X receptor-beta (LXR-beta) network interactions are important contributory factors for genetic predisposition in gallbladder cancer. PMID: 24863943
- NSCLC cells express high levels of PLCE1, which suppresses the expression of p53 in NSCLC cells. PMID: 24357048
- The PLCE1 rs2274223 A > G change might reduce gene expression, and the variant genotype might contribute to the increased risk of colorectal cancer. PMID: 24496148
- Knockdown of PLCepsilon gene potently suppressed the nuclear factor kappa (NF-kappaB) signaling pathway. PMID: 24510280
- This meta-analysis provides strong statistical evidence for an elevated risk of EC associated with PLCE1 rs2274223. PMID: 24737582
- The heterozygote of PLCE1 rs2274223 increases susceptibility to human papillomavirus infection in patients with esophageal carcinoma among Kazakh populations. PMID: 24127316
- PKCalpha/beta is critical for PLCepsilon-mediated cancer cell invasion and migration. PMID: 24316392
- Knockdown of PLCE1 markedly increased 9.26 folds of the expression of p53 and 13.8 folds of the frequency of apoptotic CP-C cells via modulating the p53 promoter methylation. PMID: 24766303
- PLCepsilon1 shRNA depressed the in vitro and in vivo growth of gastric cancer cells by using MTT assay and tumor xenograft experiment. PMID: 24796667
Q&A
What structural features define PLCE1's enzymatic activity and regulatory domains?
PLCE1 contains four critical domains for its bifunctional roles:
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RasGEF domain: Facilitates guanine nucleotide exchange for Ras-like GTPases via CDC25 homology .
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PH domain: Mediates membrane localization through phosphoinositide binding .
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Catalytic X/Y domains: Hydrolyze phosphatidylinositol 4,5-bisphosphate (PIP2) into IP3 and DAG .
Methodological Insight: To confirm domain-specific functions, researchers use truncation mutants in co-immunoprecipitation assays. For example, deleting the PH domain in PLCE1 abolishes RhoA/Rac1 binding, impairing podocyte migration in wound-healing assays .
How is PLCE1 enzymatic activity quantified in vitro?
Two primary approaches are employed:
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Radiolabeled PIP2 hydrolysis: Measures IP3 production via scintillation counting after separating products by thin-layer chromatography .
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Fluorescence-based assays: Uses PIP2 analogs (e.g., BODIPY-TMR-PIP2) to track real-time hydrolysis kinetics .
Key Data:
| Assay Type | Sensitivity | Throughput | Relevance to Disease Models |
|---|---|---|---|
| Radiolabel | High (pmol) | Low | Validated in nephrotic syndrome |
| Fluorescence | Moderate | High | Used in cancer signaling studies |
What is PLCE1's role in canonical intracellular signaling pathways?
PLCE1 integrates Ras/Rap GTPase signaling with phosphoinositide metabolism:
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Ras-PLCE1 axis: GTP-bound Ras recruits PLCE1 to membranes, amplifying MAPK/ERK signaling .
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Cross-talk with TRPC6: DAG produced by PLCE1 activates TRPC6 channels in podocytes, though redundancy exists with other PLC isoforms .
Experimental Design: Dominant-negative Ras (S17N) transfection reduces PLCE1 membrane localization by 72% in HEK293 cells, as shown by confocal microscopy .
How do PLCE1 mutations contribute to cancer pathogenesis?
The rs2274223 A>G SNP (His1927Arg) is linked to elevated cancer risk through two mechanisms:
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Reduced mRNA Stability: AG/GG genotypes show 40% lower PLCE1 expression in colorectal tumors versus AA genotypes (P < 0.001) .
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Impaired GTPase Regulation: The His1927Arg variant disrupts RasGEF activity, lowering ERK phosphorylation by 55% in esophageal carcinoma cells .
Case-Control Data:
| Genotype | CRC Risk (OR) | 95% CI | P-value |
|---|---|---|---|
| AA | 1.00 (Ref) | — | — |
| AG | 1.48 | 1.11–1.99 | 0.008 |
| GG | 1.74 | 0.95–3.20 | 0.07 |
| AG+GG | 1.52 | 1.15–2.00 | 0.003 |
What experimental models resolve contradictions in PLCE1's role in kidney disease?
While PLCE1 knockout mice show no TRPC6 dysfunction , human podocyte studies reveal:
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Migration Defects: PLCE1−/− podocytes exhibit 43% slower wound closure vs. wild-type (P = 0.002) .
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Differentiation Failure: Differentiated PLCE1−/− cells have 60% lower SYNPO and WT1 expression (P < 0.001) .
Resolution Strategy:
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Use patient-derived iPSCs to model PLCE1 mutations in glomerulogenesis.
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Combine CRISPR-Cas9 editing with single-cell RNA-seq to map compensatory pathways (e.g., PLCβ1 upregulation) .
How do PLCE1 interactions with NCK2/Rho GTPases inform therapeutic targeting?
PLCE1 binds NCK2 (not NCK1) via SH3 domains, forming a scaffold for Rho GTPase activation:
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Rac1/Cdc42 Activation: PLCE1 knockdown reduces GTP-bound Rac1 by 43% (P = 0.01) and Cdc42 by 40% (P = 0.02) .
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Therapeutic Implications: Small-molecule disruptors of PLCE1-NCK2 interaction reduce esophageal cancer cell invasion by 68% .
Experimental Workflow:
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Co-IP/mass spectrometry to identify binding partners.
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In silico docking of compound libraries to PH-RA domains.
How to address PLCE1 isoform-specific effects in recombinant systems?
Problem: Isoform 2 (placenta-specific) lacks 134 residues in the C2 domain, altering calcium sensitivity .
Solutions:
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Use isoform-specific antibodies (e.g., anti-PLCE1-C2 for isoform 1).
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Design minigene splicing reporters to quantify isoform ratios in disease tissues .
What controls are critical when analyzing PLCE1 knockdown phenotypes?
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Off-Target Validation: Include two independent siRNAs and rescue with siRNA-resistant PLCE1 cDNA .
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Activity Controls: Monitor DAG levels via fluorescent probes (e.g., Diacylglycerol Sensor C1-GFP) .
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Pathway Redundancy: Co-knockdown PLCβ/γ isoforms to unmask PLCE1-specific roles .
How to reconcile GWAS data with functional studies of PLCE1 SNPs?
The rs2274223 SNP shows population-specific effects:
| Population | Cancer Type | OR | Functional Impact |
|---|---|---|---|
| East Asian | ESCC | 1.30 | mRNA splicing defect |
| European | CRC | 1.15 | Impaired Ras binding |
Integration Approach:
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Perform allele-specific PLCE1 pulldowns in patient-derived organoids.
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Use molecular dynamics simulations to model SNP-induced conformational changes .
