Recombinant Mouse Acyl-protein thioesterase 1 (Lypla1) represents a full-length protein expressed in E. coli, with an N-terminal GST tag that makes purification more straightforward. SDS-PAGE analysis indicates purity levels above 90%, though this protein is intended strictly for research purposes. Scientists may find this a dependable resource for investigating enzyme activity and protein interactions across different experimental conditions.
Acyl-protein thioesterase 1 (Lypla1) appears to play an important role in controlling protein palmitoylation dynamics—a post-translational modification that influences where proteins end up in cells and how they function. This enzyme seems to be connected to multiple cellular processes, particularly signal transduction and membrane trafficking. Understanding how this protein works could be key to grasping how protein interactions and functions get fine-tuned in living systems.
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 Lypla1 is expressed in E. coli, a prokaryotic system that may be suitable for this cytosolic enzyme, as it does not require complex post-translational modifications. The protein is full-length (1-230aa) with high purity (>90%), and the GST tag may enhance solubility. However, since activity is unverified, the Lypla1 protein cannot be assumed to be correctly folded or bioactive. Lypla1 requires precise folding of its α/β hydrolase domain for enzymatic activity, and while E. coli can often fold soluble cytosolic proteins; experimental validation is essential.
1. GST Pull-Down Assays for Protein-Protein Interaction Studies
The GST tag enables pull-down experiments to identify potential binding partners. However, if Lypla1 is misfolded, interactions may not be physiological. The high purity reduces background but does not guarantee native conformation. This application should include proper controls and validation of the folding state before biological interpretation.
2. Antibody Development and Validation
This application is appropriate. The recombinant Lypla1 can serve as an effective immunogen for generating antibodies. The full-length sequence ensures broad epitope coverage, and the GST tag facilitates purification. Even if misfolded, antibodies may still recognize linear epitopes useful for techniques such as Western blotting.
3. Biochemical Characterization and Enzyme Kinetics Studies
This application requires validation first. While the Lypla1 protein can be used for biochemical studies, enzyme kinetics assays are only valid if the protein is properly folded and active. Initial studies should focus on verifying thioesterase activity before quantitative kinetic analysis.
4. Comparative Species Analysis
This application is feasible but requires caution. Comparative studies are only meaningful if the recombinant Lypla1 protein is properly folded. If misfolded, species differences may be obscured by folding artifacts. This should be pursued after confirming proper folding and activity.
Final Recommendation & Action Plan
Given the uncertainty in folding and bioactivity, recommend first performing functional validation through enzymatic activity assays using known Lypla1 substrates (e.g., palmitoylated peptides). Conduct biophysical characterization (size exclusion chromatography, circular dichroism) to assess folding state. Once activity is confirmed, the recombinant Lypla1 protein can be used for all the described applications. For immediate use, antibody development is the safest application. Always include appropriate controls and validate findings with complementary methods when possible.
