SPP 1128:  Optical Analysis of the Structure and Dynamics of Supra Molecular Biological Complexes

• Subject Area: Biology
• Term: from 2003 to 2010
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 5471435

The goal of the programme is the quantitative optical analysis and theoretical modelling of the three dimensional structure and dynamics of supra molecular complexes of biological macromolecules. These "molecular machines" shall be examined especially in the context of the relevant cellular structures as they are of fundamental importance for numerous cellular functions. In the centre of interest are quantitative highest resolution light optical methods of analysis in connection with specific fluorescence labelling methods and quantitative approaches of interpretation by the use of modelling and simulation. Together with these methods of analysis complementary analysis of ultra-structures are appreciated.The interdisciplinary works is concentrated on the following topics:1. Membrane associated and cytoplasmatic complexes e.g. the optical analysis and modelling of coatameric complexes; structure analysis of the centrosome.2. Genesis and function of the synaptic cytomatrix, e.g. high resolution analysis of transport of proteins and protein complexes into the synapses.3. Active chromatin complexes e.g. optical analysis and modelling of the nanostructure of specific human gene regions; optical analysis of chromatin remodelling complexes in living cells.It is intended to locally establish methods for high resolution light optical procedures, fluorescence information procedures, and light optical nanoscopy, and make them available to support the investigations.

• DFG Programme: Priority Programmes
• International Connection: Israel

Projects

• 3D Analysis of Nuclear Nanostructures by Spatially Modulated Illumination Microscopy (Applicant Cremer, Christoph )
• Analysis of the in vivo chromatin conformation by high resolution fluorescence microscopy (Applicant Rippe, Karsten )
• Analysis of VDAC multi-protein complexes with dual-color fluorescent protein nanoscopy (Applicant Jakobs, Stefan )
• Complex regulatory dynamics of the actin cytoskeleton - a new approach combining photoactivation and microfluidic tools (Applicant Beta, Carsten )
• Coordinator grant (Applicant Cremer, Christoph )
• Determination of protein associations within the human inner kinetochore in living cells using Acceptor Bleaching FRET and FLIM (Applicant Diekmann, Stephan )
• Dynamic Simulation of active/inactive Chromatin Domains (Dynamische Simulation aktiver/inaktiver Chromatindomänen) (Applicant Kreth, Gregor )
• Dynamics of Chromatin Accessibility and Proteome Distribution in Living Cells (Applicants Cardoso, Maria Cristina ;  Leonhardt, Heinrich )
• Dynamics of membrane associated actin-Arp2/3 and myosin-I complexes (Applicant Gerisch, Günther )
• Dynamik der Chromatinzugänglichkeit und der Proteomverteilung in lebenden Zellen (Applicant Leonhardt, Heinrich )
• Electron cryo-tomography of excitatory synapses and the structural characterisation of the postsynaptic molecular complexes (Applicant Baumeister, Wolfgang )
• Elucidating the intracellular mechanisms of RNA interference by single molecule fluorescence microscopy and spectroscopy (Applicant Schwille, Petra )
• Employing Fluorescence Saturation for the Functional and Spatial Optical Analysis of Cellular Supramolecular Complexes (Applicant Heintzmann, Rainer )
• Evaluation of different models of nuclear architecture based on the quantitative optical analysis of compaction levels of functionally defined chromatin regions (Applicant Cremer, Thomas )
• Functional analysis of synaptic cytomatrix proteins by novel, minimal-invasive Picosecond-FLIM and Picosecond-Anisotropy Fluorescence Imaging Microscopy (Applicant Zuschratter, Werner )
• Functional dynamics and molecular modelling of the promyelocytic leukemia (PML) nuclear body (Applicant Hemmerich, Peter )
• Functional dynamics and molecular modelling of the promyelocytic leukemia (PML) nuclear body (Applicant Langowski, Jörg )
• Imaging assembly and turnover of the presynaptic cytomatrix (Applicant Dresbach, Ph.D., Thomas )
• Imaging the dynamics of actin-associated proteins in motile cells using ultra-fast, multi-spectral TIRF microscopy (Applicant Diez, Stefan )
• Investigation of structure and function of the COPI-System using single, on nanostructures immobilised molecules and fluorescence resonance energy transfer (FRET) (Applicant Wieland, Felix Wilhelm Theodor )
• Investigation of structure and function of the COPI-System using single, on nanostructures immobilised molecules and fluorescence resonance energy transfer (FRET) (Applicant Gölzhäuser, Armin )
• Localisation and visualisation of biological markers and tracking of structures in living cells (Applicant Hesser, Jürgen )
• Mechanics and fast conformational dynamics of the nuclear pore complex (Applicant Florin, Ernst-Ludwig )
• Molecular and Microscope Analysis of Dictyostelium Microtubule Plus End Complexes and Their Role in Microtubule/Cell Cortex Interactions (Applicant Gräf, Ralph )
• Molecular interactions of a receptor/transducer complex during photosignaling studied with single molecule fluorescence spectroscopy (Applicant Fitter, Jörg )
• Molecular mechanisms of SNARE-patterning (Applicant Lang, Thorsten )
• Observation of single transport events through single pore complexes of the nuclear envelope by means of near-field optical microscopy (Applicant Naber, Andreas )
• Optical accessibility analysis of macromolecule complexes to active/inactive chromatin regions studied by fluorescent beads in live cell nuclei (Applicant Cremer, Christoph )
• Optical analysis of the oxygen-regulated assembly of hypoxia inducible factor-1 (HIF-1) by means of 2 photon laser FRET and FRAP (Applicants Fandrey, Joachim ;  Metzen, Eric )
• Single molecule fluorescence studies of the structure and dynamics of chromatin complexes (Applicants Langowski, Jörg ;  Seidel, Claus )
• Single molecule fluorescence studies of the structure and dynamics of chromatin complexes (Einzelmolekül-Fluoreszenzuntersuchungen zur Struktur und Dynamik von Chromatinkomplexen) (Applicant Seidel, Claus )
• Threedimensional optical analysis of hypoxia inducible factor-1 (HIF-1) activated nuclear gene complexes by means of 2 P Confocal Laser Scanning and Spatially Modulated Illumination Nanoscopy, and 2P Spectral Precision Distance Microscopy (Applicant Metzen, Eric )
• Threedimensional optical analysis of hypoxia inducible factor-1 (HIF-1) activated nuclear gene complexes by means of 2 P Confocal Laser Scanning and Spatially Modulated Illumination Nanoscopy, and 2P Spectral Precision Distance Microscopy (Applicant Cremer, Christoph )
• Translocation of single molecules by the pore complex of the nuclear envelope analyzed by low-light level three-channel fluorescence microscopy (Applicant Kubitscheck, Ulrich )
• Using dynamic light microscopy and cryo-electron tomography we will investigate the actin cytoskeleton during gliding motility of malaria parasites (Applicant Frischknecht, Friedrich )
• Visualization and quantitative evaluation of molecular and celluar structures by processing data obtained from cryo-electron tomography (Applicant Hegerl, Reiner )

• Spokesperson: Professor Dr. Christoph Cremer