Quantitative methods for the analysis of the spatial and temporal dynamic changes of the proteome during host-pathogen interaction

Based on the progress achieved in the area of mass spectrometry, proteomics, the large-scale study of proteins, has evolved into a powerful approach that enables quantification of several thousand proteins from a microorganism on a global scale. Proteomics is therefore one of the keystones of systems biology approaches which describe host-pathogen interactions. Project Z2 provides a sophisticated proteome analysis platform to specific A (Aspergillus) and C (Candida) projects of the CRC/TR FungiNet. The portfolio of proteome techniques includes 2D-gel electrophoresis and LC-MS/MS-based shotgun proteomic methods for identifying and quantifying the proteome of an organism on the level of peptides (“bottom-up” approach). Furthermore, we have established a serological proteome analysis method for the characterization of fungal protein antigens. In this funding period, our aim is to broaden the portfolio of proteomic methods. These include metabolic labeling, the investigation of different post-translational modifications (phosphorylation, acetylation, ubiquitination, protein thiol oxidation), absolute quantification of proteins based on quantifier peptides and subcellular fractionation (for example vesicles) to improve the dynamic range of proteome analyses. With MALDI MS Imaging (MSI) we are now able to analyze the spatial distribution of low molecular weight compounds like secondary metabolites, toxins and proteins in the interaction of the fungal pathogen and the host.