Welcome to our platform for binary protein-protein interaction screening and validation with the automated yeast-2-hybrid (Y2H) and the LUMIER systems.
Prof Erich E. Wanker
Over the last 10 years the research group Neuroproteomics has established an automated platform for the identification and validation of protein-protein interactions, enabling affordable and efficient high-throughput screenings. This platform is available to our colleagues at the MDC and other research organizations for their projects.
The platform is operated by highly experienced researchers and technicians and is constantly extended and updated. Besides the classical yeast-two-hybrid (Y2H) system (Fields und Song 1989, Nature) we have also established the MYTH Y2H system (split-ubiquitin based membrane yeast-two-hybrid, Stagljar et al. 1998, Proc. Natl. Acad. Sci. U.S.A.) and the cytoY2H system (cytosolic yeast-two-hybrid; Yeast split-ubiquitin-based cytosolic screening system to detect interactions between transcriptionally active proteins. Mockli, N., et al., 2007, Biotechniques). MYTH allows die identification of interactions between integral membrane proteins and membrane-associated/proximal proteins. CytoY2H enables the detection of PPIs in the cytoplasm.
In addition to these detection methods, we also offer a well established method for the validation of identified interactions. The LUMIER assay (Barrios-Rodiles et al. 2005, Science) is based on the isolation and co-purification of two tagged proteins from transiently transfected mammalian cells. Like the screening assays, the LUMIER assay has been adapted to automated robotics protocols.
The detection and validation units facilitate the discovery of protein-protein interactions, the confirmation of putative interactions and, more generally, new insights into the function of target proteins. For the interaction screenings, a prey-library arrayed in microtiter plates is available. It contains ~23.000 entry-clones, of which ~14.000 are unique full-length cDNAs. This library covers more than 75% of the human genome. As a special service, we are able to re-array the library according to your needs and interests.