TGFBR2 Human, His
Description
TGFBR2 is expressed with a 239aa hIgG-His tag at C-Terminus and purified by proprietary chromatographic techniques.
Product Specs
Q&A
Basic Research Questions
What structural features of TGFBR2 Human, His are critical for experimental design?
TGFBR2 Human, His is a 43.3 kDa recombinant protein expressed in Sf9 Baculovirus cells, containing a 239-amino acid hIgG-His tag at the C-terminus . Key structural elements include:
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Extracellular domain: Binds TGF-β ligands (e.g., TGF-β1).
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Intracellular serine/threonine kinase domain: Mediates downstream SMAD phosphorylation .
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Transmembrane region: Facilitates heterodimerization with TGFBR1 .
Methodological consideration: Use SDS-PAGE (40–57 kDa migration range) and Western blotting with anti-His antibodies to confirm protein integrity .
How does TGFBR2 function in canonical TGF-β signaling?
TGFBR2 initiates signaling by binding TGF-β ligands, recruiting TGFBR1 to form a heterotetrameric complex. This activates TGFBR1’s kinase domain, leading to SMAD2/3 phosphorylation, complex formation with SMAD4, and nuclear translocation to regulate gene expression . Experimental validation: Monitor SMAD2 phosphorylation (e.g., via Western blot) in cell lines (e.g., HCT116) after TGF-β1 stimulation .
What disease models are commonly used to study TGFBR2 dysfunction?
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Colorectal cancer: HCT116 TGFBR2-reconstituted cell lines (doxycycline-inducible) model microsatellite instability (MSI) tumors .
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Marfan syndrome (MFS): Transgenic mice or patient-derived fibroblasts with mutations (e.g., M425V, R460H) in the kinase domain .
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Aortic aneurysms: TGFBR2 knockout models to study vascular smooth muscle cell dysregulation .
Advanced Research Questions
How do researchers resolve contradictory data on TGFBR2’s role in cancer progression?
TGFBR2 exhibits dual roles: tumor-suppressive in colorectal cancer (via SMAD-mediated growth arrest) and oncogenic in oral squamous cell carcinoma (via non-canonical pathways) . Analytical approach:
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Context-specific signaling analysis: Compare SMAD2 phosphorylation vs. MAPK activation in different cell types.
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Glycosylation profiling: Use lectin arrays to assess TGFBR2-induced sialylation changes in MSI vs. microsatellite-stable tumors .
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Transcriptomic screens: RNA-seq to identify TGFBR2-regulated genes in organ-specific models.
What methodological challenges arise when reconstituting TGFBR2 in MSI cell lines?
Key challenges include:
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Transgene silencing: Mitigated by single-copy integration (e.g., Flp-In system) and doxycycline-inducible promoters .
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Protein stability: TGFBR2 peaks at 6 hours post-induction but degrades by 48 hours; use cycloheximide chase assays to quantify half-life .
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Functional validation: Confirm kinase activity via SMAD2 phosphorylation assays under TGF-β1 stimulation .
How are TGFBR2 mutations analyzed for pathogenicity in aortic disorders?
A tiered workflow is recommended:
What techniques identify TGFBR2-interacting proteins in signalosome studies?
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Co-immunoprecipitation (Co-IP): Use anti-His magnetic beads to pull down TGFBR2 complexes from cell lysates .
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Proximity ligation assays (PLA): Visualize real-time TGFBR2-TGFBR1 interactions in fixed cells .
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Crosslinking mass spectrometry: Identify transient interactors (e.g., endoglin) in vascular endothelial cells .
Data Contradiction Analysis
Why do TGFBR2 expression levels vary between tumor types?
Discrepancies arise from:
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Epigenetic regulation: Promoter hypermethylation in gastric cancer vs. hypoxia-induced upregulation in breast cancer .
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Post-translational modifications: Ubiquitination-mediated degradation in colorectal cancer vs. stabilisation by chaperones in fibroblasts .
Resolution strategy: Perform paired analyses of mRNA (qPCR) and protein (Western blot) levels across ≥3 cell lines per tumor type.
How do TGFBR2 mutations differentially affect connective tissue vs. cancer phenotypes?
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Marfan-associated mutations (e.g., M425V): Impair kinase activity, leading to SMAD2 hypoactivation and ECM dysregulation .
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Cancer-associated mutations (e.g., frameshifts in polyA tracts): Cause truncated receptors that dominantly inhibit wild-type TGFBR2 .
Experimental design: Use isogenic cell lines (wild-type vs. mutant TGFBR2) and transcriptomic profiling to map pathway-specific effects.
Product Science Overview
Transforming Growth Factor Beta Receptor II (TGFBR2) is a crucial component of the Transforming Growth Factor Beta (TGF-β) signaling pathway. This pathway plays a significant role in regulating various cellular processes, including proliferation, differentiation, and apoptosis. The recombinant form of TGFBR2, tagged with a histidine (His) tag, is commonly used in research to facilitate purification and detection.
TGFBR2 is a serine/threonine kinase receptor that binds to TGF-β ligands. Upon ligand binding, TGFBR2 forms a complex with Transforming Growth Factor Beta Receptor I (TGFBR1), leading to the phosphorylation and activation of TGFBR1. This activation triggers downstream signaling cascades, primarily through the SMAD pathway, which regulates gene expression and cellular responses .
The recombinant production of TGFBR2 involves the insertion of the TGFBR2 gene into an expression vector, which is then introduced into host cells, such as E. coli or mammalian cells. The His tag is added to the N- or C-terminus of the protein to facilitate purification using immobilized metal affinity chromatography (IMAC). This method allows for the efficient isolation of the recombinant protein from the host cell lysate .
Recombinant TGFBR2, His Tag (Human) is widely used in various research applications, including:
- Structural studies: Understanding the three-dimensional structure of TGFBR2 and its interactions with ligands and other receptors.
- Functional assays: Investigating the role of TGFBR2 in cellular signaling and its impact on cellular processes.
- Drug discovery: Screening for potential inhibitors or modulators of TGFBR2 activity, which could be used in therapeutic interventions for diseases related to TGF-β signaling .
