Recombinant Human Protein Wnt-2b (WNT2B)
Description
Introduction to Recombinant Human Protein Wnt-2b (WNT2B)
Recombinant Human Protein Wnt-2b (WNT2B) is a secreted signaling molecule belonging to the Wnt family, which regulates critical biological processes such as embryonic development, tissue regeneration, and cellular homeostasis . This protein is synthesized recombinantly to study its roles in canonical Wnt/β-catenin signaling and therapeutic potential in diseases like cancer, liver fibrosis, and neurodegenerative disorders .
Protein Properties
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Expression Systems:
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Post-Translational Modification: Requires palmitoleoylation for binding to frizzled receptors; depalmitoleoylation inhibits signaling .
Mechanism of Action
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Canonical Pathway: Activates β-catenin-dependent signaling by binding frizzled receptors .
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Non-Canonical Roles: Modulates mitochondrial function in neurons and inhibits TLR4/NF-κB pathways in hepatic stellate cells (HSCs) .
Developmental Biology
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Embryonic Lung Development: Plays a redundant role in branching morphogenesis .
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Intestinal Regeneration: Epithelial WNT2B is upregulated after E. coli EspP-induced injury and is essential for colonoid regeneration .
Disease Models
Product Comparison
| Supplier | Catalog # | Host | Purity | Applications |
|---|---|---|---|---|
| Abcam | ab132538 | Wheat germ | >90% | WB, ELISA |
| Cusabio | CSB-CF856613HU | E. coli | >90% | SDS-PAGE, functional assays |
| Origene | TP762265 | E. coli | >80% | Cell culture, binding studies |
Liver Fibrosis
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Mechanism: Wnt2b inhibits TLR4 signaling in HSCs, reducing collagen deposition and NF-κB/MAPK activation .
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Outcome: Hydrodynamic injection of Wnt2b-overexpression plasmids attenuated fibrosis in murine models .
Neuroprotection
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Aβ-Induced Damage: Recombinant Wnt2b (300 ng/mL) restored mitochondrial membrane potential and reduced ROS in HT22 hippocampal cells .
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Key Pathways: Enhanced BDNF expression and suppressed GSK3β hyperactivation .
Challenges and Future Directions
Product Specs
Note: If you have specific requirements regarding the glycerol content, please indicate them when placing your order.
For lyophilized powder delivery forms, the buffer used before lyophilization is a Tris/PBS-based buffer containing 6% Trehalose.
This recombinant Human WNT2B protein encompasses amino acids 1-391. The calculated molecular weight of the WNT2B protein is 59.8 kDa. Expression of this WNT2B protein is conducted in an in vitro e.coli expression system. An N-terminal 6xHis-SUMO tag has been fused into the coding gene segment of WNT2B, facilitating the detection and purification of the WNT2B recombinant protein during subsequent expression and purification stages.
The protein Wnt-2b (WNT2B) is a subject of extensive research, with primary areas of investigation spanning developmental biology, cell signaling, and tumorigenesis. In developmental biology, WNT2B, as a member of the Wnt family, plays a pivotal role in embryonic development and organ formation, particularly in the development of the nervous system and muscle tissues. In cell signaling, WNT2B regulates various cellular functions, including proliferation, differentiation, and migration, through the Wnt signaling pathway. In the context of tumor development, the aberrant expression of WNT2B is associated with the occurrence and progression of multiple cancers, making it a significant target in cancer biology research.Note: We will prioritize shipping the format currently in stock. However, if you have a specific format preference, please specify it when placing your order, and we will accommodate your request.
Generally, the shelf life for the liquid form is 6 months at -20°C/-80°C. The shelf life for the lyophilized form is 12 months at -20°C/-80°C.
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.
Target Background
- Wnt2B collaborates with Frizzled7 to mediate mesenchymal to epithelial transition in colorectal cancer. PMID: 28560804
- Altered WNT2B expression in bladder wall fibroblasts modifies the expression of the apoptosis inductor TNFSF10. PMID: 29131138
- We identified two independent SNPs (i.e., WNT2B rs1175649 G>T and BTRC rs61873997 G>A) exhibiting a predictive role in CM-specific survival, with an effect-allele-attributed hazards ratio (adjusted hazards ratio) of 1.99 (95% confidence interval = 1.41-2.81, P = 8.10 x 10(-5)) and 0.61 (0.46-0.80, 3.12x10(-4)), respectively. PMID: 28499756
- A549-Luc-positive cells demonstrated cytotoxicity following exposure to the Ad-shWnt2B vector. The percentage of viable cells was significantly lower in A549-Luc cells treated with Ad-shWnt2B compared to Ad-scramble (p<0.01 versus control or Ad-scramble, respectively). PMID: 27793913
- ACSL5 mediates antiproliferative activities via Wnt2B palmitoylation with reduced Wnt activity. The molecular pathway is likely relevant for intestinal homeostasis, potentially overridden by other pathways in carcinogenesis. PMID: 25356045
- Findings suggest that the high-expression levels of Gli1 and Wnt2B may play a pivotal role during tumorigenesis of pancreatic cancer PMID: 25120849
- Results indicate that miR-185-3p contributes to radioresistance of nasopharyngeal carcinoma via modulation of WNT2B expression. PMID: 25297925
- Secretion of WNT2B and WNT9B and stabilization of beta-catenin (CTNNB1) upon virus infection negatively regulate the expression of representative inducible genes IFNB1, IFIT1, and TNF in a CTNNB1-dependent effector mechanism PMID: 23785285
- High Wnt2B overexpression is associated with ovarian cancer metastasis and drug resistance. PMID: 22635028
- This is the first report on differential regulation of WNT2 and WNT2B mRNAs in human cancer cell lines. PMID: 11712082
- Wnt-2b mRNA is expressed in human ovarian cancer cells PMID: 12072409
- REVIEW: Anti-WNT2B monoclonal antibodies, WNT2B RNAi compounds, or small molecule WNT2B inhibitors could be developed as novel therapeutic agents for gastric cancer and esophageal cancer in the field of clinical oncology. PMID: 16273293
- In addition to alternative promoters and RNA splicing, an alternative translation start in Wnt13B and Wnt13C mRNAs increases the complexity of both human wnt13 expression and functions PMID: 16407296
- Heplipin induced KG-1 cell apoptosis is related to Wntl3 and ATPase3. PMID: 17649721
- Hedgehog signals and bHLH transcription factors are involved in WNT2B upregulation, which is counteracted by BMP, WNT, and Notch signals. PMID: 19360354
Q&A
What is Recombinant Human Protein Wnt-2b and what is its structure?
Recombinant Human Wnt-2b (WNT2B) is a full-length protein consisting of 391 amino acids that belongs to the Wnt family of secreted glycoproteins. It functions as a ligand for members of the frizzled family of seven transmembrane receptors and operates primarily through the canonical Wnt/β-catenin signaling pathway . The protein is also known by alternative designations including WNT13, Protein Wnt-2b, and Protein Wnt-13 .
The amino acid sequence begins with MLRPGGAEEA and continues through a series of hydrophobic and hydrophilic regions that contribute to its structural integrity and function . When expressed recombinantly, such as in wheat germ expression systems, the protein maintains its biological activity and can be used in various experimental applications including SDS-PAGE, ELISA, and Western blotting .
What are the known isoforms of WNT2B and how are they regulated?
WNT2B exists in multiple isoforms that show differential expression and regulation in tissues. Three primary isoforms have been identified:
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WNT2B1 (previously known as WNT13B)
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WNT2B2 (previously known as WNT13A)
Research on epithelial regeneration after injury has shown that these isoforms are differentially regulated. Following injury, WNT2B3 shows significant upregulation, while WNT2B2 exhibits slight downregulation, and WNT2B1 shows an upward trend . This differential expression pattern suggests specific roles for each isoform in the regenerative response. The mosaic expression pattern observed in both normal human colon crypts and undifferentiated colonoids further indicates complex spatial regulation of WNT2B expression .
How is WNT2B distributed in normal intestinal tissue?
WNT2B exhibits a distinctive expression pattern in normal intestinal tissue. Immunostaining and RNAscope analysis have revealed that WNT2B expression is concentrated in specific, rare epithelial cells within the human colon crypt . This expression follows a mosaic pattern, with varied amounts of WNT2B present from crypt to crypt, suggesting functional heterogeneity across the intestinal epithelium .
In longitudinal sections of colonic tissue, WNT2B mRNA is also detected in mesenchymal cells of the lamina propria, consistent with previous reports highlighting the importance of mesenchymal Wnt production . This dual source of WNT2B from both epithelial and mesenchymal compartments likely contributes to the complex regulation of intestinal homeostasis and regeneration processes .
What methods can be used to study WNT2B function in intestinal regeneration models?
Several experimental approaches have been developed to investigate WNT2B function in intestinal regeneration:
Colonoid Injury Models: Researchers have established in vitro models using enterohemorrhagic E. coli-secreted cytotoxin EspP to induce epithelial injury in human colonoids. This approach provides a simplified system that lacks mesenchymal Wnts, allowing for the specific study of epithelial-derived WNT2B in regeneration .
Recombinant Protein Supplementation: Recombinant human WNT2B (rhWNT2B) can be added to colonoid cultures simultaneously with Wnt pathway inhibitors like IWP-2. This approach has demonstrated that rhWNT2B alone is sufficient to rescue and promote regeneration after injury, providing direct evidence of its functional importance .
Mouse Knockout Models: Wnt2b knockout (KO) mice provide an in vivo system to study the consequences of WNT2B deficiency. These models allow for assessment of baseline histology and health of both small intestine and colon, as well as the impact of inflammatory challenges using agents like dextran sodium sulfate (DSS) .
How does Recombinant Human WNT2B contribute to regeneration after intestinal injury?
Recombinant Human WNT2B plays a critical role in promoting regeneration following intestinal epithelial injury. In experimental models using human colonoids injured with EspP toxin, the addition of rhWNT2B was sufficient to rescue regeneration even in the presence of the Wnt pathway inhibitor IWP-2 . This finding demonstrates that WNT2B alone can drive the regenerative process independent of other Wnt ligands.
The mechanism appears to involve stimulation of stem cell proliferation and reorganization of the epithelium. Studies in other systems, such as chick retinal explants, have shown that Wnt2b overexpression leads to increased cell proliferation and the growth of large, folded tissue structures . This proliferative activity is likely mediated through the canonical Wnt/β-catenin pathway, as WNT2B functions as a ligand for frizzled receptors, ultimately leading to transcriptional activation of target genes .
Notably, the palmitoleoylation of WNT2B is required for its efficient binding to frizzled receptors, with depalmitoleoylation leading to inhibition of the Wnt signaling pathway . This post-translational modification represents an important regulatory mechanism for WNT2B activity during regeneration.
What is the relationship between Desert Hedgehog (DHH) signaling and WNT2B expression?
A significant interplay exists between Desert Hedgehog (DHH) signaling and WNT2B expression in the context of intestinal regeneration. Research has demonstrated that DHH acts as a driver of regeneration and specifically modulates WNT2B expression .
This relationship suggests a coordinated signaling network where Hedgehog pathway activation, specifically via DHH rather than Indian or Sonic Hedgehog, upregulates WNT2B expression to promote epithelial regeneration after injury. This finding highlights the importance of cross-talk between major developmental signaling pathways in adult tissue repair processes.
What are the phenotypic consequences of WNT2B deficiency in experimental models?
WNT2B deficiency leads to significant phenotypic consequences, particularly related to intestinal function and inflammation susceptibility:
Human WNT2B Deficiency: Individuals with WNT2B deficiency present with severe intestinal disease, including significant inflammatory injury, highlighting WNT2B's critical role in intestinal homeostasis .
Mouse Knockout Models: Interestingly, Wnt2b knockout (KO) mice show more subtle baseline phenotypes. Examination of intestinal architecture in Wnt2b KO mice revealed similar structure to wild-type mice under normal conditions, with normal distribution of Paneth cells, goblet cells, and brush border in the epithelium .
Gene Expression Changes: Despite normal histology, Wnt2b KO mice exhibit approximately 40% reduction in Lgr5 expression (a marker for intestinal stem cells) compared to control mice, while the expression of differentiated lineage markers remains similar . This suggests that WNT2B may regulate intestinal stem cell function without dramatically affecting differentiated cell lineages under homeostatic conditions.
Inflammatory Susceptibility: The most significant phenotype of WNT2B deficiency appears to be enhanced susceptibility to colitis, indicating that WNT2B plays a critical role in protecting against intestinal inflammation . This increased vulnerability to inflammatory damage may result from compromised epithelial regenerative capacity during inflammatory challenge.
How can organoid models be utilized to investigate WNT2B function?
Organoid models offer powerful tools for investigating WNT2B function in intestinal epithelium:
Establishment and Maintenance: Small intestinal organoids (enteroids) can be established from both wild-type and Wnt2b knockout mice using standard organoid culture media lacking exogenous Wnt ligands . Despite the absence of WNT2B, Wnt2b KO enteroids show similar morphology to wild-type controls and can be expanded for more than 15 passages, indicating normal growth and cell division capacity .
Injury and Recovery Assessment: Organoids can be subjected to various injury models to assess the role of WNT2B in regeneration. For example, enteroids can be treated with interferon-γ (IFNγ), which causes Paneth cell destruction and organoid death . Following injury, recovery can be monitored by assessing organoid survival, morphology, and budding crypt formation.
WNT2B Dependence Studies: While Wnt2b KO enteroids can grow in culture, studies with human colonoids derived from WNT2B-deficient individuals have shown that these cultures are not stable and unable to form robust cultures . This discrepancy highlights potential species-specific differences and compensatory mechanisms.
Recombinant WNT2B Supplementation: The addition of recombinant WNT2B to organoid cultures allows for the assessment of rescue effects. This approach has demonstrated that recombinant murine WNT2B allows for the formation of short-term enteroid cultures from WNT2B-deficient patients .
What methodologies are appropriate for detecting WNT2B expression in experimental systems?
Multiple complementary approaches can be employed to detect WNT2B expression in experimental systems:
Immunostaining: Immunohistochemistry can be used to visualize the distribution of WNT2B protein in tissue sections and organoid cultures. This technique has revealed that WNT2B expression is concentrated in specific, rare epithelial cells in normal human colon crypts and undifferentiated colonoids .
RNAscope: This in situ hybridization technique offers highly sensitive detection of WNT2B mRNA in tissues. RNAscope analysis has shown that WNT2B is localized to the deep crypt in human colonic tissue and is present in a mosaic pattern with varied expression levels from crypt to crypt .
Quantitative Real-Time PCR (qRT-PCR): This method can detect and quantify WNT2B mRNA expression, even at low levels. Studies have shown that WNT2B mRNA is detectable in enteroids at low levels (qRT-PCR cycle count 35-37), supporting reports that the epithelium can produce WNT2B, albeit at minimal levels .
Proteomics Screening: Proteomics approaches have successfully identified differential expression of WNT2B isoforms, such as the upregulation of WNT2B3 in EspP-injured regenerating colonoids .
What are the key translational implications of WNT2B research?
Research on WNT2B has significant translational implications for understanding and treating intestinal diseases. The enhanced susceptibility to colitis observed in WNT2B-deficient models suggests that WNT2B plays a protective role against inflammatory bowel diseases . This finding opens potential therapeutic avenues for treating conditions characterized by intestinal inflammation.
The ability of recombinant WNT2B to rescue regeneration in experimental models highlights its potential as a therapeutic agent for promoting intestinal repair after injury . This could be particularly relevant in clinical scenarios involving intestinal damage, such as radiation enteritis, infectious colitis, or inflammatory bowel disease.
Furthermore, the understanding of WNT2B's role in intestinal stem cell biology may inform approaches to intestinal tissue engineering and regenerative medicine. The observation that WNT2B is essential for the stability and propagation of human colonoid cultures underscores its importance in maintaining stem cell populations , knowledge that could be leveraged for developing cell-based therapies.
