Participant 21: PLUS
Paris-Lodron-Universitaet Salzburg, Fachbereich Molekulare Biologie, Austria
Participant leader: Prof. Dr. Christian Huber
Relevant experience and laboratory resources:
The group of Prof. Christian Huber at the University of Salzburg has extensive experience in the development and application of multidimensional chromatographic separation methods hyphenated to mass spectrometry for protein and proteome analysis. Introducing novel monolithic separation media, it was possible to fractionate peptide mixtures with previously unknown separation efficiency before identification by electrospray ionization- (ESI) or matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). The implementation of alternative schemes for two-dimensional separation of complex peptide mixtures (reversed-phase x ion-pair reversed-phase chromatography) facilitated a significant improvement in proteome coverage and identification confidence. In collaboration with Bruker Daltonics, Bremen, the potential of electron transfer dissociation (ETD) is investigated aiming at improved peptide identification and the detection of post-translational modifications. Using this novel fragmentation technology in combination with the high resolving power of monolithic columns for large peptides and intact proteins, the applicability of this approach to top-down proteome analysis is critically evaluated. The group collaborates closely with Prof. Knut Reinert (FU Berlin) and Prof. Oliver Kohlbacher (Universtiy of Tübingen) in the development of bioinformatic tools for high-throughput protein identification and quantification (OpenMS). A major focus of this work lies in the increase of reproducibility and confidence of protein identification for proteome analysis. Using retention time prediction models, we aim at utilizing information inherent to the analytical process for eliminating false positive peptide identifications. Laboratory resources at University of Salzburg include all necessary equipment for sample preparation of proteomes and metabolome, instruments for multidimensional chromatography, ESI- as well as MALDI-mass spectrometers for analyte identification.
Principal scientific personnel involved:
Prof. Dr. Christian Huber
Dr. Hanno Stutz
Role of the participant in the project and main tasks:
The participant will contribute to WP4 by conducting differential metabolome/cellome analyses of normal and treated cell systems.