300402 UE Yeast Genetics and Proteomics Tools (2013S)

In this laboratory course we will present methods that are required to prepare samples of sufficient quality for proteomics/MS analysis. All of the presented techniques are based on specific affinity tags and therefore we generate yeast strains by endogenous tagging of the the genes of interest. We will apply protocols for the tandem affinity purifications of whole protein complexes to study the complex composition, and also individual proteins to study their PTMs. Purified samples will be separated on SDS-PAGE and analyzed by mass spectrometry compatible gel staining techniqes. Additionally, we will demonstrate the advantages/disadvantages of a novel system to study transient protein-protein interactions called M-Track.