Recombinant Human Glutaminyl-peptide cyclotransferase (QPCT) is produced in a mammalian cell expression system, which appears to ensure proper folding and post-translational modifications. The protein includes the full length of the mature protein, covering amino acids 29-361, and features an N-terminal 10xHis-tag for simplified purification and detection. SDS-PAGE analysis confirms the protein purity exceeds 90%, which likely makes it suitable for various research applications.
Glutaminyl-peptide cyclotransferase (QPCT) is an enzyme that participates in post-translational modification processes. It catalyzes the formation of pyroglutamate from glutaminyl residues at the N-terminus of peptides and proteins. This modification may influence protein stability and function. QPCT holds significance in research related to protein maturation and degradation pathways, and its activity proves relevant in studies focusing on enzyme mechanics and protein biochemistry.
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.
1. Biochemical Characterization and Enzyme Kinetics Studies
Researchers can use this recombinant QPCT protein to investigate the fundamental biochemical properties of glutaminyl-peptide cyclotransferase. These studies might examine substrate specificity, optimal reaction conditions, and kinetic parameters. The high purity (>90%) and mammalian expression system appear to ensure proper protein folding and post-translational modifications that could be critical for enzymatic function. Scientists can establish in vitro assays using various peptide substrates to determine catalytic efficiency and identify preferred substrate sequences. The N-terminal His-tag allows for easy purification and immobilization for continuous assays or biophysical studies.
2. Protein-Protein Interaction Studies
The His-tagged QPCT can work as bait protein in pull-down assays to identify novel binding partners or validate known interactions with other cellular proteins. The tag allows efficient immobilization on nickel-affinity matrices for capturing interacting proteins from cell lysates or purified protein libraries. This approach may help clarify the cellular networks and regulatory mechanisms involving QPCT in various biological processes. The mammalian expression system likely ensures native-like protein conformation that preserves physiologically relevant binding interfaces.
3. Antibody Development and Validation
This purified recombinant protein works as an ideal immunogen for generating specific antibodies against human QPCT, including both polyclonal and monoclonal antibodies. The high purity level minimizes cross-reactivity with contaminating proteins. Meanwhile, the mammalian expression system provides native protein structure for generating antibodies that recognize the natural form of QPCT. Scientists can also use the protein to validate antibody specificity through ELISA, Western blot, and competitive binding assays. The His-tag allows easy coating of ELISA plates and provides an additional epitope for detection if needed.
4. Structural Biology and Biophysical Analysis
Researchers can use the recombinant QPCT protein for structural studies including X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy to understand the three-dimensional architecture of the enzyme. The mammalian expression system increases the likelihood of obtaining properly folded protein suitable for structural analysis. Biophysical techniques such as dynamic light scattering, analytical ultracentrifugation, and thermal stability assays can provide insights into protein oligomerization state, stability, and conformational changes. The His-tag helps with protein purification to the high concentrations typically required for structural studies.
5. In Vitro Assay Development and Screening Applications
Scientists can use this QPCT protein to develop and optimize in vitro enzymatic assays for research applications, including the establishment of standardized protocols for measuring cyclotransferase activity. The consistent quality from mammalian expression makes it suitable as a positive control or reference standard in comparative studies. Researchers can use this protein to screen for potential modulators or inhibitors in compound libraries, providing a foundation for chemical biology studies. The His-tag allows easy integration into automated screening platforms through immobilization or tag-based detection methods.
