Ubiquitination Analysis Service

The analysis of ubiquitination plays a crucial role in understanding post-translational modifications, directly influencing protein activity and localization. This modification is linked to the degradation of a substantial portion of eukaryotic proteins, approximately 80-85%, and is intricately involved in diverse cellular processes. At Creative Proteomics, we offer comprehensive ubiquitination analysis services that encompass the identification, quantification, and characterization of protein ubiquitin modifications. These services provide valuable insights into your protein research endeavors.

What is Ubiquitination

Ubiquitin, a small protein present in most eukaryotic cells, acts as a tag for proteins destined for breakdown and hydrolysis. As a protein labeled with ubiquitin enters the protease barrel, the protease initiates the hydrolysis process. This ubiquitin labeling of proteins is termed ubiquitination and is typically achieved through the formation of an isopeptide bond between the carboxyl group of ubiquitin's last amino acid and the ε-amino group of the substrate lysine. The ubiquitination process involves several sequential steps.

Fig.1 Schematic of ubiquitination

First, ubiquitin undergoes activation through a two-step reaction involving the ubiquitin-activating enzyme E1, with ATP serving as an energy source. Initially, ubiquitin reacts with ATP to form the ubiquitin-adenylate complex. Subsequently, ubiquitin is separated from AMP and transferred to E1. Then, E1 facilitates the transfer of the activated ubiquitin to the ubiquitin-binding enzyme E2 via a transsulfuration esterification reaction. Next, the final step in ubiquitination involves the transfer of ubiquitin bound to E2 to the target protein. This crucial step is carried out by E3 ligases, which recognize substrates and interact with both E2 and the substrate. Some E3 ligases also possess the ability to activate E2 activity.

Applications of Ubiquitination Analysis

In addition, ubiquitination analysis also has applications in disease mechanisms, drug discovery, immune response, cell cycle regulation and stem cell biology, etc.

Our Service

Creative Proteomics has advanced protein detection technologies and can provide customized services to researchers and scientists to help them reveal the complexity of PTM using powerful mass spectrometry technologies. Our services are thoughtful and detailed and the process of ubiquitination analysis service is as follows:

Fig.2 Ubiquitination analysis workflow

• Sample preparation
  Proper extraction and preparation of proteins from biological samples to ensure preservation of ubiquitinated proteins.
• Ubiquitination enrichment
  Isolation of ubiquitinated proteins or peptides using enrichment techniques. This step is critical because the abundance of ubiquitin modifications is typically lower compared to unmodified proteins.
• Detection
  WB, protein microarrays and LC-MS/MS are powerful tools for the identification and quantification of ubiquitinated peptides, and they provide detection information that facilitates a comprehensive analysis of ubiquitination.
• Bioinformatics analysis
  Advanced bioinformatics tools are used to analyze data, identify ubiquitinated peptides, and map these modifications to protein sequences.
• Result delivery
  After professional data analysis, we deliver results quickly and consistently.

Creative Proteomics brings a wealth of experience to the field of protein research. Our comprehensive services and products cover the entire spectrum of utilizing suitable and efficient gateways. Utilizing our proprietary platform, we have successfully implemented many proteomics research projects. If you are interested, please contact us for more information and a quote.