EMBO - Characterisation of post-translational modifications in cellular signalling

Protein post-translational modification (PTM) is a crucial mechanism in cellular regulation. Many proteins can be modified through the addition or removal of chemical groups at specific locations to alter the function of the protein in ways that produce a particular outcome for the cell. In addition, PTMs of a given protein can affect other PTMs leading to distinct functions of the protein, a phenomenon termed PTM cross-talking. Linking PTM dynamics and cell biology is an expanding area and one fraught with technical difficulties. The practical course aims to provide a theoretical and practical overview of state-of-art techniques for the characterization of various PTMs, such as phosphorylation, acetylation, cysteine PTMs and glycosylation, and their cross-talking in controlling dynamic cellular mechanisms. The students will be introduced to enrichment, identification and quantitation of four different PTM peptides from complex biological matrices enabling the students to perform quantitative large scale PTMomics from small amount of sample material. The course will provide individuals with hands-on experience on both the purification of modified peptides using robust methods developed by among others the applicants laboratory and mass spectrometry analysis, as well as on bioinformatics on their own data obtained from EGF stimulated SILAC labeled HeLa cells.