Recombinant Pig Growth Hormone Receptor (GHR) is expressed in E. coli and contains amino acids 19 to 264 of the full-length protein, representing a partial sequence. This protein comes engineered with a C-terminal 6xHis-tag, which helps streamline purification and detection processes. Its purity appears to exceed 85% as confirmed by SDS-PAGE analysis, suggesting it's suitable for various research applications. The product is intended for research use only and not for diagnostic or therapeutic purposes.
The Growth Hormone Receptor (GHR) plays a critical role in regulating growth and metabolism. It participates in the growth hormone signaling pathway, mediating growth hormone effects by binding to it and triggering downstream signaling cascades. This receptor seems essential for various physiological processes, making it an important target for studies on growth disorders and metabolic research.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
This GHR fragment corresponds to the extracellular domain of GHR, which requires correct folding, including specific disulfide bond formation and potential glycosylation, for bioactivity (e.g., binding to growth hormone). E. coli, as a prokaryotic expression system, lacks the eukaryotic chaperones and post-translational modification machinery (e.g., glycosylation) necessary for proper folding of complex receptor domains. The absence of glycosylation and the potential for improper disulfide bonding make it highly unlikely that this protein is correctly folded or bioactive. The high purity indicates low impurity levels, but does not guarantee native conformation. Without experimental validation (e.g., circular dichroism for secondary structure or binding assays), the protein is probably misfolded and inactive for functional applications.
1. Protein-Protein Interaction Studies
If the recombinant GHR fragment is verified to be correctly folded (e.g., through biophysical or binding assays), it could be used to study interactions with growth hormone or ligands via techniques like surface plasmon resonance. However, if misfolded (which is likely), binding results may be unreliable due to altered binding sites or non-specific interactions. Any findings must be validated with full-length, eukaryotic-expressed GHR to ensure physiological relevance. The His tag facilitates immobilization but may contribute to artifacts.
2. Antibody Development and Characterization
This recombinant protein can be used as an immunogen for generating antibodies against linear epitopes within the GHR extracellular domain. The high purity supports consistent immunization and screening. However, antibodies produced may not recognize native, correctly folded GHR (e.g., on cell surfaces) due to conformational epitope differences. Antibody specificity should be validated against eukaryotic-expressed GHR for applications like Western blot or immunofluorescence.
3. Structural and Biochemical Analysis
The protein is suitable for basic biophysical characterization (e.g., circular dichroism to assess secondary structure, dynamic light scattering to monitor aggregation) to evaluate folding status. However, it is not appropriate for high-resolution structural studies (e.g., X-ray crystallography) or analyses assuming native conformation due to likely misfolding. These assays can help identify folding issues but not provide insights into functional structure.
4. In Vitro Competitive Binding Assays
This application requires a correctly folded protein for meaningful results. If the protein is misfolded (as expected), competitive binding assays will yield inaccurate data (e.g., false negatives/positives). Prior validation of growth hormone binding activity is essential. Without confirmation, this protein should not be used for drug screening or mechanistic studies; instead, use eukaryotic-expressed GHR.
Final Recommendation & Action Plan
Before using this recombinant GHR fragment for any functional application, it is critical to validate its folding state through biophysical assays (e.g., circular dichroism to check for expected β-sheet content, size-exclusion chromatography to assess oligomerization) and functional tests (e.g., growth hormone binding assays). If misfolding is confirmed, limit use to non-functional applications like antibody production, but always validate antibodies against native GHR. For reliable protein-protein interaction or binding studies, express the GHR extracellular domain in a eukaryotic system (e.g., mammalian or insect cells) to ensure proper folding and glycosylation. Avoid functional assays without prior activity confirmation.
