Recombinant Human Probable E3 ubiquitin-protein ligase HERC1 is produced in an E.coli expression system. The protein features a 10xHis-tag at the N-terminus and a Myc tag at the C-terminus. This partial protein covers amino acids 3975-4360 and reaches a purity level greater than 85%, as confirmed by SDS-PAGE analysis. It's designed for research purposes and appears to offer a solid tool for studying protein interactions and modifications, with low endotoxin levels that may be beneficial for cell-based studies.
HERC1 belongs to the ubiquitin-protein ligase family. It seems to play an important role in the ubiquitination process, which targets proteins for degradation through the proteasome pathway. This protein likely participates in various cellular pathways and processes, which makes it particularly interesting for research into protein regulation and cellular homeostasis. Studying HERC1 could potentially improve our understanding of cellular mechanisms and how proteins are managed throughout their lifecycle.
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 HERC1 is a large E3 ubiquitin ligase that requires precise folding, proper domain organization (including potential HECT domain functionality), and specific tertiary structure for its enzymatic activity. The E. coli expression system cannot provide the eukaryotic folding environment, chaperones, or post-translational modifications necessary for the correct folding of this complex mammalian protein. The partial fragment (3975-4360aa) may contain functional domains, but the dual N-terminal 10xHis and C-terminal Myc tags are likely to cause steric interference with domain folding and functional sites. The probability of correct folding with functional ubiquitin ligase activity is extremely low without experimental validation.
1. Antibody Development and Validation
This application is conditionally suitable. Antibody development relies on antigenic sequence recognition. The fragment can generate antibodies, but the immune response may primarily target the tags or linear epitopes due to potential misfolding. Antibodies may not recognize natively folded, full-length HERC1 in physiological contexts.
2. Structural and Biochemical Characterization
Basic biophysical characterization can be performed, but will not reflect native HERC1 structure. The tags will dominate physical properties, and the results will describe an artificial construct. Techniques like CD spectroscopy or DLS can assess folding state, but cannot confirm functional conformation.
Final Recommendation & Action Plan
This E. coli-expressed HERC1 fragment with dual tags is unsuitable for functional studies due to the high probability of misfolding in the prokaryotic system and severe tag interference. Application 1 has limitations for generating conformation-specific antibodies. Application 2 can only provide basic physical data on the recombinant construct. For reliable HERC1 research, use full-length or larger domain constructs expressed in mammalian systems that support proper folding and avoid tag artefacts.
