Protein-interactionscreening using quantitative mass spectrometry

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

In order to understand neurodegenerative disease networks extensive information of 
protein-protein interactions (PPIs) is essential. Genetic (e.g. Y2H) and biochemical 
approaches to investigate PPIs provide largely complementary datasets. The goal of 
this subproject is to identify PPIs involved in neurological disorders by affinity 
purification and quantitative mass spectrometry. In addition, we will investigate 
differences in PPIs between wild-type and mutant forms of selected disease proteins. 
In order to achieve this, we will develop and employ a method that combines affinity 
purification with quantitative mass spectrometry. The general idea is to 
quantitatively compare interaction partners of fusion proteins with suitable 
endogenous controls. Specific interaction partners are identified by their abundance 
ratio. By combining our dataset with results of the Y2H screens in other NeuroNet subprojects, we expect to capture the majority of the interactions of the disease proteins. Furthermore, the data on differences in interaction partners of 
wild-type and mutant forms of disease proteins will provide a direct link to disease
phenotypes.

 

 
                        

Electrospray ionisation of peptides for a LTQ-Orbitrap mass spectrometer