Establishing structure-function relationships of lipolytic enzymes and their interaction partners
In living organisms, excess energy is stored mainly as triglycerides (TG) in storage organelles termed lipid droplets or lipid bodies. In periods of increased energy demand, this stored energy is mobilized through lipolysis. Lipases are the enzymes hydrolyzing ester bonds of water-insoluble substrates such as TG, cholesteryl esters, and phospholipids. Energy homeostasis requires a fine-tuned balance between accumulation and mobilization of TG from these organelles. Aberrant function of lipases or its regulators are associated with a large variety of metabolic disorders such as obesity, cardiovascular diseases, type 2 diabetes, and certain forms of cancer. Despite the obvious importance of these proteins, structural knowledge in this field is still scarce. The group of M. Oberer focuses on the structure-function relationship of proteins involved in lipid metabolism from different organisms. The group aims at understanding the underlying molecular mechanism and protein-protein interactions on a structural level. To achieve these goals a large array of techniques is used, including molecular biology methods, biochemistry and biophysics. The group has a special emphasis on structural biology and utilizes protein crystallography and biomolecular NMR spectroscopy.
Laboratory know-how and infrastructureThis laboratory’s focus is in molecular structural biology, particularly in the study of lipases and their protein-protein interactions. The group of M. Oberer is embedded in the Structural Biology Group at the Institute of Molecular Biosciences. Competence in this laboratory includes cloning, expression and purification of proteins, biocalorimetry, and various biochemical and spectroscopic techniques. The determination of 3D crystal and solution structures is performed employing X-ray crystallography and NMR spectroscopy, respectively. The laboratories are equipped with state-of-the-art molecular biology and protein-purification facilities. A cell culture laboratory and isotope facilities are available at the institute and frequently used by the group. The local infrastructure also includes comprehensive biochemical facilities (centrifuges, incubators, FPLC and HPLC systems, chromatographic columns, electrophoresis units) and state-of-the-art instrumentation for biophysical experiments (UV/VIS, CD- and fluorescence spectroscopy, biocalorimetry, macromolecular light scattering). The Structural Biology Group is equipped with hard and software covering all aspects of biomolecular NMR spectroscopy and X-ray crystallography. Local X-ray facilities include two rotating anode generators with imaging-plate detectors and which also harbor equipment for cryogenic cooling during data collection. The crystallization laboratories are equipped with a routinely used high-throughput crystallization robot and thermoregulated crystallization closets. Local NMR facilities are used in collaboration with K. Zangger and include a Bruker Avance III 700MHz with cryogenic equipment, a Varian Unity INOVA 600, a Bruker Avance DRX 500, and a Varian Unity INOVA 400.