Phosphorylation is participating in almost all cellular processes and is the most studied post-translational modification today. Phosphoproteomics aim at characterize all phosphoproteins in a cell or tissue at a given time. Several strategies exist for phosphoproteomics, most combining peptide fractionation techniques with efficient phosphopeptide enrichment methods and mass spectrometry. The course focuses on the practical issues concerning quantitative phosphoproteomics and tips and tricks which is of outmost importance in being able to characterize phosphoproteins.

Phosphorylation is one of the most abundant post-translational modification (PTM) in nature and it participate in almost all cellular processes. Proteins gets reversibly phosphorylated in the cell by a group of enzymes termed kinases,  whereas another enzyme group, phosphatases, are able to remove this phosphate group again. Often a complex interplay takes place between these two enzyme groups and often these enzymes are themself controlled by reversible phosphorylation. In order to understand such phosphorylation-dependent regulation processes in cells in detail, efficient methods for the characterization of phosphorylated proteins are needed. Many of such methods are previously described with various success and output. However, it is well known that enrichment methods for phosphorylated peptides are of outmost importance for establishing an efficient strategy for large scale phosphoproteomics.

The course focuses on practical issues concerning many phosphoproteomics aspects, which are of outmost importance in being able to characterize phosphorylated proteins. It is aimed for scientists like cell biologists, bio-/protein-chemists and other with interest in cellular signalling and regulation where phosphorylation are key mechanisms or researchers who are heading towards implementing phosphoproteomic strategies in their research project.

The practical course aims to provide an overview of state-of-art techniques for the purification, analysis, identification and quantification of phosphopeptides in complex samples using different enrichment strategies, various tandem mass spectrometry technologies and data interpretation and validation strategies.

 It will provide individuals with hands-on experience on both the phosphopeptide purification steps and mass spectrometry analysis, as well as on data analysis within the area of phosphoproteomics.

The organizer of this course has been in the area of phosphoproteomics since 1996. He has made a significant methodological contribution to the area by inventing numerous methods for assessment of phosphorylation sites and enrichment of phosphorylated peptides ((Titanium dioxide chromatography: Larsen MR et al., Mol Cell Proteomics 2005, 4(7), 873-86; SIMAC: Thingholm TE et al., Mol Cell Proteomics. 2008, 7(4), 661-71); large scale methods: Engholm-Keller K et al., J Proteome Res. 2011 Dec 2;10(12):5383-97; Engholm-Keller K et al. J. Proteomics. 2012 Oct 22;75(18):5749-61. ).





Department of Biochemistry & Molecular Biology, University of Southern Denmark
Campusvej 55
DK-5230 Odense M, Denmark

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