NGFN-PLUS
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
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
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