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Protein-protein interactions play an integral role in numerous cellular processes. Membrane protein interactions, in particular, are attracted more and more attention because of their role in disease and because of their prevalence as major pharmaceutical targets. However, the hydrophobic nature of membrane proteins has made them difficult to study in a high-throughput format. Creative Biogene, as one of the leading biotechnology companies, is able to provide membrane protein Y2H library screening service to help you discover novel membrane protein interactions effectively and accurately.
Figure 1. Schematic of membrane protein Y2H screening.[3]
Membrane protein yeast two-hybrid screening is based on the split-ubiquitin protein complementation assay which allowing the use of full-length integral membrane proteins and membrane-associated proteins as baits to hunt for novel interaction partners. Membrane protein Y2H screening takes advantage of the observation that ubiquitin can be separated into two stable moieties: the C-terminal half of yeast ubiquitin (Cub) and the N-terminal half of the ubiquitin moiety (Nub). In briefly, your bait protein is fused to Cub moiety which is linked to an artificial transcription factor and the prey proteins (either in individual or library format) are fused to the Nub moiety. When bait protein and prey proteins interact, it will lead to reconstitution of the ubiquitin moieties, forming a full-length 'pseudo-ubiquitin' molecule. This molecule is in turn recognized by cytosolic deubiquitinating enzymes, resulting in cleavage of the transcription factor, and subsequent initiate the transcription of the reporter genes. Using the characterization of the reporter gene, we can finally investigate interactions of integral membrane proteins from both yeast and other organisms.
Applications:
Creative Biogene has extensive expertise and experience which can provide you with comprehensive membrane protein Y2H library screening service to identify proteins that interact with a protein of your interest using your pre-made library or our custom-built Y2H library. Creative Biogene possesses sophisticated equipments, advanced technologies, highly experienced staffs which are able to provide you personalized screening services to meet your analysis needs. Please feel free to contact us. We are looking forward to working together with your attractive projects.
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