NR AURF
AU del Pino,P.; Weiss,A.; Bertsch,U.; Renner,C.; Mentler,M.; Grantner,K.; Fiorino,F.; Meyer-Klaucke,W.; Moroder,L.; Kretzschmar,H.A.; Parak,F.G.
TI Molecular Structure of the Cu2+ Binding Sites of Full-Length Human Prion Protein
QU TSE-Forum, 6. Kongress - Nationale TSE-Forschungsplattform, Greifswald 26.6.-28.6.2006, Poster: Struktur und molekulare Mechanismen MOL-03
PT Konferenz-Poster
AB Copper ions bind under physiologically relevant conditions to the octapeptide repeat region of the cellular prion protein PrPc. Several studies have investigated PrPc-derived peptides lacking the structured C-terminal part of the protein to determine the molecular features of the copper binding sites. In the present study, the correctly folded human full-length PrPc (hPrP 23-231) was loaded under slightly acidic (pH 6.0) or neutral conditions with 63Cu. PrPc-Cu2+-complexes precipitated under non-denaturing conditions were investigated by EXAFS, EPR, and ENDOR. Guided by these results we were able to build a model for the Cu2+ complex using molecular mechanics computations to select sterically possible and energetically favourable structures. To verify our model for the Cu2+ complex we compared the simulated EPR, ENDOR, and EXAFS spectra of these structures with our experimental data. The investigations presented here show the importance of the application of different experimental methods to resolve ambiguity in the individual results. Our spectroscopic data on shorter PrPc-derived peptides shows the marked influence of sequence context, buffer conditions and amount of copper on the molecular features of the Cu2+ complex.
AD Pablo del Pino, Andreas Weiss, Christian Renner, Matthias Mentler, Klaus Grantner, Fritz G. Parak, Physik Department E17, Technische Universität München, 85747 Garching; Ferdinando Fiorino, Luis Moroder, Laboratory of Bioorganic Chemistry, Max-Planck-Institut für Biochemie, 82151 Martinsried; Uwe Bertsch, Hans A. Kretzschmar, Zentrum für Neuropathologie und Prionforschung der LMU, 81377 München; Wolfram Meyer-Klaucke, EMBL Outstation Hamburg
SP englisch
PO Deutschland
OR Tagungsband