Mating-based Split-Ubiquitin System (mbSUS)

Mating-based Split-Ubiquitin System (mbSUS)

Introduction

Reversible and dynamic protein-protein interactions play a vital role in all living cells, so it has become an increasingly popular research project. In order to gain a deeper understanding of the interactome, scientists have developed a mating-based split-ubiquitin system (mbSUS), which is a sensitive and user-friendly alternative to the classic yeast two-hybrid system and has been successfully used for many analyses of protein interactions. mbSUS relies on the ubiquitin-degradation pathway as a sensor, and is very suitable for detecting the interaction of full-length proteins localized in the cytoplasm and in or at membrane compartments, without their truncation and nuclear mislocation.

When the N-terminal domain of ubiquitin (Nub) and its C-terminal half (Cub) are co-expressed, functional ubiquitin can be reconstructed. Compared to wild-type Nub, the Nub mutant NubG (containing mutation Ile13Gly) has a reduced affinity for the Cub moiety and thus cannot reconstruct the functional ubiquitin unless it is brought into the vicinity of the Cub domain through the interaction of the two fusion partners. The split ubiquitin system was further improved by introducing a mating approach (mbSUS) for high-throughput screening. In short, mbSUS provides an efficient and multi-functional tool for protein-protein interaction research, which is a supplement to BiFC and FRET technologies.

The mbSUS for detection of protein-protein interactionsFigure 1. The mbSUS for detection of protein-protein interactions. USP: ubiquitin specific protease. (Grefen, C.; et al. 2007)

Services

Mating-based-Split-Ubiquitin-System-(mbSUS)-2

Creative Proteomics established the mbSUS platform many years ago and recruited a large number of experienced experts to manage the platform. Using this technology platform, we can provide customers with feasible, easy-to-use and cost-effective strategies. Under the conventional experimental procedures, we can also perform operations such as adding reporter genes and promoters and optimizing vectors according to customer needs. Our experimental process includes but is not limited to: 

  • Yeast transformation
  • Testing the bait fusion for background activity
  • Mating
  • Quantitative and qualitative detection

Customers can choose different technology platforms according to project requirements, or contact us directly for consultation, and our expert team will provide you with customized experimental procedures.

Creative Proteomics is an international biotechnology company dedicated to research in molecular interactions and other related fields. The mating-based split-ubiquitin system we constructed has the characteristics of high quality and efficiency, and the data obtained can be directly used for paper publication. Our one-stop service aims to save customers time and money.   

References

  1. Obrdlik, P.; et al. K+ channel interactions detected by a genetic system optimized for systematic studies of membrane protein interactions. PNAS. 2004.
  2. Grefen, C.; et al. Split-Ubiquitin System for Identifying Protein-Protein Interactions in Membrane and Full-Length Proteins. Current Protocols in Neuroscience. 2007.
  3. Gronemeyer, T.; et al. A Split-Ubiquitin Based Strategy Selecting for Protein Complex-Interfering Mutations. G3. 2016.
* This service is for RESEARCH USE ONLY, not intended for any clinical use.

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