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Harald Pichler

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Harald Pichler
Institute of Molecular Biotechnology
Technische Universität Graz
Petersgasse 14
8010 Graz

e-Mail: harald.pichler@tugraz.at
phone: 0043 316 873 4089
fax: 0043 316 873 4071
web: www.tugraz.at

Molecular interactions governing membrane-associated enzyme function

The lipid constituents of biological membranes, as well as membrane-associated or membrane-embedded proteins and protein complexes determine many essential cellular functions. Although recent “-omics” approaches have discerned the importance of single membrane components, only limited knowledge of the molecular events at and within membranes is available. Nevertheless, the last years have seen a few outstanding developments in our understanding of membrane-related processes. For example, it is now known how mammalian cells use protein - lipid molecular interaction to sense their cholesterol status and regulate cholesterol biosynthesis and uptake in response (Goldstein et al., 2006). Membrane protein - protein interaction was demonstrated to inhibit the ratelimiting step of sphingolipid biosynthesis in S. cerevisiae (Breslow et al., 2010; Han et al., 2010). Last but not least, highly specific sterol - sphingolipid interactions can be key determinants of enzyme activities suggesting that membrane-attached enzymes may be commonly regulated by their lipid and protein surroundings (Guan et al., 2009). To elucidate protein - lipid interactions governing membrane protein function on the molecular level, the key issue will be to identify and characterize domains or even single amino acids of membrane proteins as necessary and/or sufficient to respond to the presence or absence of defined lipid species.

Laboratory know-how and infrastructure

The group is experienced in the detailed analysis of processes involving biological membranes, particularly regarding intracellular localization and transport as well as lipid and protein composition. Lately, a second focal point has been recombinant protein expression and enzyme engineering as the team is strongly involved in the Austrian Center of Industrial Biotechnology (ACIB), which has evolved from the Applied Biocatalysis (A-B) Research Centre. Being housed in the same building, the Institute of Molecular Biotechnology and ACIB share equipment that is state-of-the-art in molecular biosciences. We have standard facilities for molecular biology and yeast research, i.e. cell culture (< 5 liter), diverse imaging systems (fluorescence and chemiluminescence), and infrastructure for cell harvest, cell fractionation and protein purification. Also in house we have a Biacore system for molecular interaction studies, the latest generation of capillary electrophoresis of DNA/RNA/protein samples, and (HP)LC-MS and GC-MS analytical instruments. Furthermore, tools for genetic manipulations and high-throughput screening techniques on the colony level are established, including workstations for automated colony picking and liquid handling.

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