Recombinant Human MGAT4A is expressed in E.coli and covers amino acid region 93-535, representing a partial length version of the protein. The product includes an N-terminal 6xHis tag for easier purification and detection. SDS-PAGE analysis confirms the protein reaches purity levels above 85%. This preparation is designed for research use only and appears to deliver consistent performance across different experimental setups.
MGAT4A represents a key glycosyltransferase that modifies N-glycans and plays an important role in creating complex-type oligosaccharides. The enzyme's activity affects N-glycan branching patterns, which may influence protein folding and cell-cell interactions. Studying MGAT4A could help researchers better understand glycosylation pathways and how they relate to cellular function and disease processes.
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.
Human MGAT4A is a Golgi-resident glycosyltransferase that requires precise folding, disulfide bond formation, and proper membrane association for its enzymatic activity in N-glycan biosynthesis. The E. coli expression system cannot provide the necessary eukaryotic post-translational modifications, disulfide bond formation, or membrane environment critical for this enzyme's native conformation and function. The partial fragment (93-535aa) represents the catalytic domain but lacks the N-terminal cytoplasmic and transmembrane regions. While the protein may be soluble, it is highly unlikely to achieve the correct folding needed for functional glycosyltransferase activity.
1. Biochemical Characterization
Basic biophysical characterization (e.g., thermal stability, pH tolerance) can be performed regardless of folding state. However, functional enzymatic characterization requires proper folding that E. coli cannot provide.
2. Antibody Development and Validation
This recombinant MGAT4A serves as an excellent immunogen for generating antibodies against linear epitopes. The partial sequence provides substantial epitope coverage. However, antibodies may not efficiently recognize conformational epitopes on the native, properly folded enzyme.
Final Recommendation & Action Plan
The E. coli expression system is fundamentally unsuitable for producing a functional version of this complex Golgi glycosyltransferase. The recombinant MGAT4A fragment is primarily suitable for antibody development (Application 2) and basic biophysical characterization (Application 1), but requires rigorous validation before any functional applications. Begin with basic biophysical characterization to assess folding status through techniques like circular dichroism spectroscopy and size-exclusion chromatography. Application 2 (antibody development) can proceed immediately for linear epitope antibodies. If activity validation is attempted and fails, limit applications to non-functional uses only.
