Institut für Pathologie

Medizinische Universität Graz

Forschungseinheit für
Functional Proteomics and Metabolic Pathways

Scientific Focus:
The scientific experience of the research unit “Functional Proteomics and Metabolic Pathways” lies within lipid metabolism, molecular and cell biology, enzymes, protein analytics and proteomics. Our laboratory aims at global analysis of protein function in basic medicine and biotechnology with a special focus on lipid metabolism. We have developed tools for activity-based discovery, profiling and imaging of lipases and other serine hydrolases. We investigate lipid metabolism using functional proteomic approaches for the study of lipases and their regulators, as well as lipoproteins and lipid droplets in health and disease. Currently we focus on the regulation of lipid and energy homeostasis by identifying unknown protein interaction partners and continue current efforts to study post-transcriptional modifications of lipases and their regulators. We perform analyses of the molecular (phospho)proteomic networks related to lipolysis in health and disease to increase the understanding of the dysregulation of lipid metabolism in tumor growth and heart dysfunction.

Infrastructure:
Our laboratory is equipped with several high resolution mass spectrometers: The Thermo LTQ-FT consists of a linear iontrap and a Fourier transform ion cyclotron resonance mass spectrometer, which is able to deliver MSn spectra with mass accuracies of less than 1 ppm. The faster and more sensitive Thermo Orbitrap velos pro coupled to a Dionex Ultimate RSLC nano-HPLC provides higher coverage of complex proteomic samples. The Bruker Ultraflextreme MALDI-TOF-TOF mass spectrometer provides complementary data. Additional mass spectrometers (Bruker maXisII Q-TOF, EvoQ EMR triplequadrupole and amazon speed ETD iontrap) were recently installed as part of the Omics Center Graz. For protein separation, one and two-dimensional gel electrophoresis systems, and for peptide separation one and two-dimensional liquid chromatography are used on a regular basis. Mass spectrometry-based quantitative proteomic methods, including label-free approaches, such as spectral counting and retention time alignment and quantitation of areas of reconstructed ion chromatograms or targeted mass spectrometry, as well as isotope labelling approaches such as dimethyllabelling, ICPL, and metabolic labelling (stable isotope labeling with amino acids in cell culture, SILAC), i-TRAQ/TMT and gel-based abundance-based protein profiling (DIGE) are well established methods. A laserscanner and a spotcutter are available for gel-based proteomic approaches. Software and servers for 2D gel image analysis (Delta2D), database search, PTM and quantitative data analysis and data annotation and comparison are installed (Mascot, Proteinscape, Proteome Discoverer, Sieve, Peaks etc). Standard laboratory equipment for biochemistry and small scale organic synthesis is available. Our group is located at the Center for Medical research (ZMF), where we have access to well-equipped general and specialized laboratories (e.g. radionuclide laboratory, S2 and S3 cell culture facilities), as well as to high end technologies, such as confocal laser scanning microscopy, electron microscopy, flow cytometry/sorting, microarrays, digital PCR and next generation sequencing.

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