Characterisation of subcellular compartments that contain signalling complexes of protein kinase C epsilon

• Applicant: Dr. Simone Radtke
• Subject Area: Biochemistry
• Term: from 2005 to 2009
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 14474456

Members of the family of protein kinase C (PKC) serine/threonine kinases serve as important downstream effectors of various signalling pathways and are implicated in the regulation of diverse processes governing cellular fate and function, such as proliferation, differentiation, survival, adhesion and motility. The aim of this project is to study in detail the contribution of the epsilon PKC isoform (PKCs) to signal transduction mediated by receptor tyrosine kinases such as epidermal growth factor receptor and c-met, the receptor for the hepatocyte growth factor. These receptors become rapidly internalised upon ligand binding and PKCs has been implicated in the regulation of ensuing endosomal signalling events. We will make use of a PKCe construct that due to a mutation in its kinase domain is sensitive to specific inhibition by a chemically modified kinase inhibitor. Specific inhibition of PKCs kinase activity in growth factor stimulated cells should allow us to effectively accumulate signalling complexes containing activated PKCs in the relevant intracellular compartments. These will be isolated using cell fractionation techniques and analysed by two-dimensional polyacrylamide gel electrophoresis in order to identify PKCs interaction partners and substrates. Since dysregulation of PKCs and growth factor receptors has strong implications in the development of cancer, our findings will be of great interest for better understanding the underlying mechanisms.

• DFG Programme: Research Fellowships
• International Connection: United Kingdom
• Host: Professor Dr. Peter Parker