NR AWBU
AU Bellon,A.; Kimberlin,C.R.; Solforosi,L.; Abalos,G.C.; Cruite,J.T.; Saphire,E.O.; Williamson,R.A.
TI Structural studies of the prion replicative interface
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions S-01
PT Konferenz-Poster
AB Neurodegenerative prion diseases such as Creutzfeldt-Jakob disease and bovine spongiform encephalopathy have been closely linked to the conversion of normal cellular prion protein (PrPc) to an alternate, misfolded conformation called PrPsc. A high-resolution structure of PrPsc would provide invaluable insight into the mechanism by which the conversion process occurs. However, the natural tendency for PrPsc to self-assemble into aggregates hinders structural studies. Therefore, only low resolution structure models of PrPsc are available so far. Historically, antibody Fab-protein complexes have been used successfully to facilitate the crystallization and structural analysis of molecules that do not crystallize alone. Here Fab can serve to create protein-protein contacts different from those made by the antigen itself, or may effectively bury certain antigenic determinants, thereby reducing the propensity for unwanted aggregation. Antibody containing a heavy-chain complementary-determining region 3 (HCDR3) sequence graft of PrP 89-112 polypeptide binds specifically to PrPsc with tight affinity. Our aim is to co-crystallize 89-112 grafted Fab fragments with misfolded PrP and resolve the structure of this complex. Fab fragments of this PrPsc-specific antibody have been prepared for crystallographic analysis. Microfluidic free-interface-diffusion screens yielded crystals of Fab 89-112 that diffract to 2.7Å with an R sym of 8.6% belonging to space group P212121. Data refinement is currently underway, and will yield the first X-ray crystal structure of PrP sequence containing an N-terminal PrPsc binding motif.
AD Anne Bellon (abellon@scripps.edu), Laura Solforosi (lsolf@scripps.edu), Gil C. Abalos, Justin T. Cruite, R. Anthony Williamson (anthony@scripps.edu), Department of Immunology, The Scripps Research Institute, La Jolla, California 92037, USA; R.H. Kimberlin, Scrapie and Related Diseases Advisory Service (SARDAS), 27 Laverockdale Park, Edinburgh EH13 QQE, Midlothian, Scotland, United Kingdom
SP englisch
PO Italien