NR AOQC
AU Basaiawmoit,R.V.; Perbandt,M.; Fimmen,L.; Clos,J.; Bredehorst,R.; Notbohm,H.; Voelter,W.; Klaucke,W.M.; Svergun,D.I.; Betzel,C.
TI Structural Investigations on the Human Cellular Prion Protein and Selected Fragments
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - BR-87
PT Konferenz-Poster
AB Prion disease is caused by an aberrantly folded form of the prion protein, PrPsc, which is predominantly beta sheeted in contrast to the predominantly alpha helical, non-pathogenic varicant form, as verified by spectroscopic data. The insolubility of PrPsc has so far precluded high-resolution structural analysis and is largely attributed to the highly disordered and flexible N-terminal domain. To investigate the structure, dynamics and aggregation of prion protein, we applied different analytical techniques. As the N-terminal domain contains four sequential copies of the highly conserved octarepeat sequence PHGGGWGQ, spanning residues 60-91, which selectively binds cupric ions in vivo, we performed Extended X-ray Absorption Fine Structure (EXAFS) experiments to analyse the binding behaviour as well as the copper co-ordination in detail. The co-ordination environment of copper at different conditions will be presented. In parallel, we applied Circular Dichorism (CD), Dynamic Light Scattering (DLS) and Small Angle X-Ray Scattering (SAXS) techniques to analyse the conformational behaviour and the dynamics of aggregation of the native full-length prion protein. Using the measured radius of gyration, the potential shape of aggregates from PrPc molecules in solution was estimated at moderate resolution. For these investigations, various selected synthetic peptides were used as well as the full-length prion protein. Results will be presented accordingly. The DLS method and CD measurements were used to check the stability and behaviour of the protein and peptides under different solution conditions and served also as an effective control after the refolding procedure. We will highlight the use of DLS technique as an effective investigative tool to analyse and characterise the prion protein.
AD R.V. Basaiawmoit, M. Perbandt, L. Fimmen, C. Betzel, Institute of Medical Biochemistry and Molecular Biology, University Hospital Hamburg-Eppendorf, Germany; J. Clos, Bernard Nocht Institute for Tropical Medicine, Hamburg, Germany; R. Bredehorst, Division of Biochemsitry and Molecular Biology, University of Hamburg, Hamburg, Germany; H. Notbohm, Institute for Medical Molecular Biology, University of Lübeck, Lübeck, Germany; W. Voelter, Division of Physical Biochemistry, University Institute of Physiological Chemistry, Tübingen, Germany; W.M. Klaucke, D. Svergun, EMBL Outstation Hamburg, Hamburg, Germany
SP englisch
PO Deutschland
EA Poster-Photo und pdf-Datei