Protein ubiquitination is a dynamic multifaceted post-translational modification (PTM) that is involved in a wide range of cellular processes, including cell cycle progression, DNA damage response, endocytosis, cell signaling, autophagy, and protein quality control system. Creative BioMart's extensive experience in post-translational modification of proteins based on computational and experimental methods enables us to provide protein ubiquitination service to accelerate your project.

Ubiquitination is a process that involves the covalent attachment of 76 amino acid protein ubiquitin to protein substrates, linking to target proteins through the sequential action of ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E2), and ubiquitin ligase (E3). The most typical role of the ubiquitination pathway is the degradation of cellular proteins by the 26S proteasome. Ubiquitination of proteins leads to changes in important properties of substrate proteins, including their activity, cellular localization, interactions with other proteins, and half-life in cells. Thus, ubiquitination regulates a large number of important cellular processes, and dysfunction of ubiquitin signaling has been implicated in the development of many serious diseases, including cancer, neurodegeneration, immune disorders, and susceptibility to infection. Therefore, it has become a new target for new drug development.

Fig 1. Protein-engineering platforms for manipulating the activities of ubiquitination and its transferring cascade. (Zhao B, et al., 2020)

Services

The diversity of ubiquitination chain connectivity and length poses a great challenge to the study of protein ubiquitination and deubiquitination. The recent boom in protein engineering approaches has produced engineered proteins as powerful tools for studying protein ubiquitination-mediated cell signaling. Creative BioMart has established a professional ubiquitinated protein engineering service platform to develop more efficient methods to unravel the complex mechanisms of ubiquitin signaling mediated by different types of polyubiquitin chains. These methods are generally applicable to multiple proteins for polyubiquitination. We provide the following services for protein ubiquitination, including but not limited to:

• Probing the activity of specific ubiquitinated transferases: we have successfully developed protein engineering tools to generate reactively functional ubiquitinated conjugates.
• Manipulating ubiquitin transfer through protein engineering: by manipulating the reactivity of ubiquitination with enzymes, we can potentially control the ubiquitination status of specific proteins in cells and modulate their degradation, interactions, transport, and enzymatic activities.
• Identification of E3 substrates through protein engineering: we used affinity-based methods such as co-immunoprecipitation, yeast two-hybrid screening, and protein microarrays to directly assess E3-substrate interactions. In addition, ubiquitination transferases with tailored activities were designed based on protein engineering to recognize cellular substrates of specific E3s.

Approaches for Analyzing Protein Ubiquitination

• Unnatural amino acid incorporation.
• Expressed protein-linked protein hemisynthesis.
• High-throughput selection for phage and yeast cell surface display.
• Bioorthogonal method.
• Chemical ubiquitination method.
• Immunodetection technology based on specific antibodies.
• Radiolabeling technique.

Our mission is to provide global customers with the most comprehensive and professional customized protein ubiquitination service. If you have any special requirements about our services, please feel free to contact us. We are looking forward to working together with your attractive projects.

Reference

1. Zhao B, Tsai YC, Jin B, et al.. (2020) Protein Engineering in the Ubiquitin System: Tools for Discovery and Beyond. Pharmacol Rev. 72(2): 380-413.

• Protein Acetylation Service
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• Protein Nitrosylation Service
• Protein Phosphorylation Service
• Protein Proteolytic Service
• Protein Sulfation Service