NR AWMT
AU Rutishauser,D.; Calella,A.M.; Polymenidou,M.; Moss,R.; Mertz,K.; Aguzzi,A.
TI Proteomic analysis of the prion protein in vivo
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 CE-43
PT Konferenz-Poster
AB PrP is thought to be a signaling molecule, yet none of the proteins mediating its signals have been identified. Also, the difficulties in achieving prion replication under chemically defined conditions suggest a role for unidentified host components. We have developed transgenic mice expressing a PrP-mycTag fusion. PrPmyc rescues Shmerling's syndrome in mendelian crosses, suggesting that it is physiologically functional, and supports prion replication. We are isolating the protein complexes that co-precipitate with PrPc and/or PrPsc. Towards that goal, we have generated antibodies, POM2 and POM3, with extremely high affinities to well-defined PrP epitopes. POM2 and POM3 discriminate between PrPc and PrPsc, and are specifically eluted by distinct epitope-mimetic peptides. PrPc and PrPsc are then recaptured with antibodies to the myc tag, and eluted with myc-mimetic peptide. These unique proteomic tools allow us to confirm the identified interactors and to avoid the risk of losing possible interactions that occur in the same region of antibody-epitope. PrP-associated proteins were purified and are currently analyzed by isotope-coded affinity tags, 1D SDS-PAGE and mass spectrometry.
AD Institute of Neuropathology, University Hospital Zürich, Switzerland. E-mail: annamaria.calella@usz.ch
SP englisch
PO Italien