Recombinant Mouse Pancreatic secretory granule membrane major glycoprotein GP2 (Gp2) is produced in E. coli. The protein includes the full length of the mature sequence, spanning amino acids 22 to 531. An N-terminal 6xHis-SUMO tag has been added, and purification reaches levels exceeding 90% as verified by SDS-PAGE. This product is intended for research use only and does not contain detectable levels of endotoxin.
GP2 appears to be a major glycoprotein found in pancreatic secretory granule membranes. The protein likely plays an important role in normal pancreatic function. GP2 seems to be involved in zymogen granule membrane processes and may participate in the sorting and exocytosis of digestive enzymes. Research focused on pancreatic function and related disorders has shown considerable interest in this protein.
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.
Based on the provided information, the recombinant mouse GP2 protein cannot be assumed to be correctly folded or bioactive without experimental validation. Several critical concerns remain: 1) GP2 is a pancreatic glycoprotein that requires N-linked glycosylation for proper folding, stability, and function - modifications that cannot be achieved in the E. coli expression system; 2) The SUMO tag, while beneficial for solubility, may still interfere with the native protein structure or oligomerization state; 3) The protein lacks the N-terminal signal peptide (cleaved in mature GP2) but the bacterial expression may not support proper disulfide bond formation essential for glycoprotein structure. The >90% purity indicates minimal contaminants, but doesn't guarantee correct folding. Without validation through circular dichroism, size-exclusion chromatography, or functional assays, the protein's structural integrity and bioactivity remain uncertain.
1. Antibody Development and Validation Studies
This recombinant GP2 can generate antibodies, but they may primarily recognize linear epitopes or the SUMO tag rather than conformational epitopes of native, glycosylated GP2. The lack of glycosylation means antibodies might not recognize physiological GP2 in pancreatic tissues. For reliable antibodies, validate against native GP2 from mouse pancreatic extracts and consider using eukaryotic-expressed GP2 for immunization. The high purity reduces contamination risks but doesn't ensure antigen quality.
2. Protein-Protein Interaction Studies
The His-SUMO tagged GP2 is unsuitable for interaction studies without folding validation. The SUMO domain may cause non-specific binding or mask authentic interaction interfaces. If misfolded, identified partners may be artifacts. Validate folding first and include rigorous controls (tag-only protein, properly folded standards). Prefer mammalian-expressed GP2 for biologically relevant interactions.
3. Biochemical Characterization and Enzymatic Assays
Basic biochemical characterization (molecular weight confirmation, stability tests) is possible, but data won't reflect native GP2 due to a lack of glycosylation. The SUMO tag may alter hydrodynamic properties in SEC and DLS. Functional assays require demonstration of native-like folding and should be interpreted cautiously without glycosylation.
4. ELISA Development and Quantitative Assays
This recombinant GP2 can be used in ELISA development, but quantification of native GP2 may be inaccurate due to structural differences. The tags may affect antibody binding epitopes. Validate assays with native GP2 standards and confirm detection antibodies recognize glycosylated epitopes for reliable quantification.
Final Recommendation & Action Plan
To ensure reliable results: 1) Remove the SUMO tag via proteolytic cleavage (if cleavage sites are available) and purify tag-free GP2 for critical applications; 2) Validate folding through circular dichroism (expecting characteristic β-rich structure) and size-exclusion chromatography with multi-angle light scattering; 3) For antibody applications, characterize sera against native GP2 from pancreatic sources; 4) For interaction studies, use mammalian-expressed GP2 with proper glycosylation as positive control; 5) Always include appropriate controls (tag-only proteins, glycosylated standards) to account for structural differences. Consider eukaryotic expression systems for applications requiring native structure and function.
