Membrane-Based Yeast Two-Hybrid Screening

The biological functions of proteins can be predicted through the study of their interaction partners, and mapping protein interaction networks is one of the major goals of the postgenomic era. A large number of membrane proteins serve as major pharmaceutical targets, but they have long been difficult to study in a high-throughput format due to their hydrophobic nature. The classical yeast two-hybrid (Y2H) screening is mostly limited to the study of cytosolic or extracellular soluble proteins. To study the interaction of transmembrane protein interaction, Profacgen provides a specialized membrane-based Y2H (MbY2H) system. The MbY2H system is also called the split ubiquitin Y2H, whereas your bait protein will be fused to the C-terminal half of ubiquitin (Cub) and a transcription factor; and the transmembrane prey protein library is fused to the N-terminal half (Nub). Upon association of the bait and prey, a compete ubiquitin molecule is formed and cleaved by endogenous protease, releasing the transcription factor to initiate the transcription of the reporter genes. The most widely used reporter genes are the same as regular Y2H systems including His3 and LacZ.

Membrane-based yeast two-hybrid system is based on the split-ubiquitin protein complementation assay and is able to detect protein interactions directly at the membrane. It allows the use of full-length integral membrane proteins and membrane-associated proteins as baits to search for novel interaction partners.

Services

Taking advantage of Profacgen's MbY2H system and comprehensive membrane cDNA libraries from different species, we can help you efficiently and accurately discover novel membrane protein interactions. Our experimental process includes selection of bait and appropriate MYTH system, bait generation and validation, and screening. Customers can get the ultimate tailor-made solution on our membrane-based yeast two-hybrid screening platform. Our expertise guarantees high data quality, short turnaround time, and the best prices in the market.

Figure 2. MYTH pipeline (Snider, J.; et al. 2010)

Profacgen has accumulated lots of experience in yeast two-hybrid screening. Our professional technical team can provide customers with high-quality membrane-based Y2H screening and many related featured services. Our competitive prices and extensive expertise have earned us the trust of our collaborators. Contact us to find out how Profacgen could be of assistance.

References

1. Snider, J.; et al. Split-Ubiquitin Based Membrane Yeast Two-Hybrid (MYTH) System: A Powerful Tool For Identifying Protein-Protein Interactions. Journal of Visualized Experiments. 2010.
2. Snider, J.; et al. Detecting interactions with membrane proteins using a membrane two-hybrid assay in yeast. Nat Protoc. 2010.