Berlin 2005 – wissenschaftliches Programm

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AKB: Biologische Physik

AKB 200: Poster Session II

AKB 200.48: Poster

Dienstag, 8. März 2005, 17:00–19:00, Poster TU C

Mapping the Thermoplasma proteome - structural proteomics studies by free-flow electrophoresis and cryo-electron tomography — •Christine Kofler, Istvan Nagy, Stephan Nickell, Marius Boicu, and Wolfgang Baumeister — Max Planck Institut für Biochemie, Molekulare Strukturbiologie

Thermoplasma acidophilum is a thermoacidophilic archaeon whose genome is completely known. To carry out proteomic analysis on this organism we use cell lysates which are fractionated using free-flow electrophoresis (FFE). The FFE separates the cytoplasmic proteins according to their isoelectric point. Single fractions are then investigated by means of cryo-electron tomography (cryo-ET) which allows to obtain three-dimensional (3-D) structural information of vitrified biological specimens at a resolution of 2-4 nm. Additionally, the contents of the single fractions are characterised by polyacrylamide gel electrophoresis and mass spectrometry. The knowledge of the 3-D structure and the determination of the identity of different proteins will enable us to generate a library of templates which is used as an input for pattern recognition algorithms designed to search electron tomograms of whole ice-embedded cells. The final aim of these studies is to locate and quantify the different macromolecular assemblies within 3-D reconstructions of intact T. acidophilum cells.