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Screening for human ErbB3-interacting proteins using the novel membrane based yeast two-hybrid system

Redaktion

Für den Inhalt der Angaben zeichnet die Projektleitung verantwortlich.

Kooperation

Diese Rubrik wird erst seit 2010 erfasst.

Projektdaten

  • Projekt-Nr: GRS-003/00 
  • Förderbeitrag: CHF 413'000 
  • Bewilligung: 22.06.2000 
  • Dauer: 09.2000 - 01.2005 
  • Handlungsfeld:  Pilotprojekte, 1998 - 2018

Projektleitung

Projektbeschreibung

One third of all about 500’000 predicted proteins in human cells are localized to the cell membrane. Membrane proteins are involved in a wide range of essential cellular functions. Importantly, most pharmaceutical drugs developed today interfere with membrane proteins. For this reason, there is a huge demand from both academic researchers and biotech-pharma companies to gain further insight into pathways and interactions involving membrane proteins.
The goal of this project was to develop a novel yeast-based approach (named Membrane based yeast two-hybrid system (MbYTH)) that can detect interactions between human membrane proteins in a screening format. We have successfully applied this technology to the mammalian receptor tyrosine kinase ErbB3 and identified its three novel associated proteins. Beside its applications in target identification purposes, the MbYTH system can potentially be used for pharmaceutical purposes. In combination with high-throughput assays, compounds of potential therapeutic value such as small molecules, peptides and antibodies can be identified using several modifications of the MbYTH technology.

Was ist das Besondere an diesem Projekt?

The MbYTH represents a highly innovative and unique proteomic platform for elucidation of membrane protein interactions.

Stand/Resultate

We have successfully applied the MbYTH technology to the mammalian receptor tyrosine kinase ErbB3 and identified its three novel associated proteins. In addition, MbYTH was successfully applied to yeast and plant membrane proteins.
Furthermore, on the basis of our work in the past 4 years, it became clear that, beside its applications in target identification purposes, the MbYTH system can also be used for pharmaceutical purposes. In combination with high-throughput assays, compounds of potential therapeutic value such as small molecules, peptides and antibodies can be identified using several modifications of the MbYTH technology.

Publikationen

- Reinders, A., Schultze, W., Thaminy, S., Stagljar, I., Fromer, W.B., and Ward, J.M. (2002) Intra- and intermolecular interactions in sucrose transporters at the plasma membrane detected by the split-ubiquitin system and functional assays, Structure 10, 763-772
- Stagljar, I. and Fields, S. (2002) Analysis of membrane protein interactions using yeast-based technologies, Trends Biochem Sci 27, 559-563
- Auerbach, D., Thaminy, S., Hottiger, M.O. and Stagljar, I. (2002) The post-genomic era of interactive proteomics: facts and perspectives, Proteomics 2, 611-623
- Auerbach, D., Galeuchet-Schenk, B., Hottiger, M.O., and Stagljar, I. (2002) Genetic approaches to the identification of interactions between membrane proteins in yeast, J Recept Signal Transduct Res 22, 473-483
- Auerbach, D., Fetchko, M..J., and Stagljar, I. (2003) Proteomics approaches for generation of comprehensive protein interaction maps, Drug Discovery Today 2, 85-93
- Thaminy, S., Auerbach, D., Arnoldo, A., and Stagljar, I. (2003) Identification of novel ErbB3-interacting proteins using the splitubiquitin membrane yeast two-hybrid technology, Genome Res 13, 1744-1753
- Scheper, W., Thaminy, S., Kais, S., Stagljar, I., and Römisch, K. (2003) Coordination of N-glycosylation and protein translocation across the ER membrane in yeast by Sss1 protein, J Biol Chem 278, 37998-38003
- Stagljar, I. (2003) Finding partner(s): emerging protein interaction technologies applied to signaling networks. Science STKE 2003, pe56
- Fetchko, M..J., Auerbach, D., and Stagljar, I. (2003) Yeast genetic methods for the detection of membrane protein interactions: potential use in drug discovery, BioDrugs 17, 413-424
- Fetchko, M. and Stagljar, I. (2004) Application of the splitubiquitin membrane yeast two-hybrid system to investigate membrane protein interactions, Methods 32, 349-362

Medienecho

keine

Letzte Aktualisierung dieser Projektdarstellung  21.12.2018