Protein interaction networks

Coordinator:		Prof. Dr. Erich Wanker
Institution:		Max-Delbrück-Centrum für Molekulare Medizin (MDC) Berlin-Buch
Homepage:		www.mdc-berlin.de

Protein-tyrosine kinases (PTKs) are important regulators of intracellular signal-transduction pathways and their activity is usually tightly controlled and regulated. Perturbation of PTK signalling by mutations and other genetic alterations results in deregulated kinase activity and cancer induction. In this sub-project, we aim at the generation of a comprehensive protein interaction network for human cytoplasmic protein-tyrosine kinases and their mutant counterparts that were identified through bioinformatics and sequencing approaches. For this we will use a robot based high-throughput yeast two-hybrid (Y2H) protein-protein interaction (PPI) approach, mass spectrometry-based proteomics and bioinformatics. The protein complex information will then be utilized for the systematic identification of modulators of PTK signalling using cell-based reporter assays. This includes systematic RNAi knock-down as well as overexpression experiments with established cell model systems in order to identify proteins that influence signalling in human malignancies.

The results of this sub-project are expected to help our partners within the MUTANOM consortium to identify new tumor suppressors or oncogenes in patient genotyping and phenotypic analyses. Furthermore, this sub-project may contribute to the elucidation of new pathways in oncogenic kinase signalling and the identification of new drug targets for therapy development.

Protein-Protein-Interaction (PPI) screening:
(a) One of the robots used for the high throughput yeast two-hybrid PPI screens, (b) an example of the PPI networks as it is generated in our lab, and (c) a plate showing some positive PPIs.



Additional relevant Internet link:
The Mutanom project