NR AXWU
AU Solforosi,L.; Bellon,A.; Abalos,G.C.; Cruite,J.; Williamson,R.A.
TI Identifying Key Components of the PrPc-PrPsc Replicative Interface
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.07
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Interaction between the cellular prion protein (PrPc) and its disease-associated conformer (PrPsc) is a fundamental event in prion replication that precedes the conversion of PrPc into nascent PrPsc. Using a large panel of recombinant antibody molecules displaying overlapping PrP peptide grafts, we have precisely identified three regions of PrPc (composed of amino acid residues 23-33, 98-110 and 136-158) that specifically and robustly interact with PrPsc. We hypothesize that these PrP sequences likely represent peptidic components of one flank of the prion replicative interface. Our experiments indicated that the reactivity of PrP-regions 23-33 and 89112 with PrPsc is in large part conferred by positively charged residues in these sequences. Within the 136-158 PrPsc binding motif, peptide sequence composed of amino acids 136-140 and 149-158 were found to be of particular importance for the binding interaction with PrPsc.
To further study the role of the PrP sequences 23-33, 98-110 and 136-158 in PrPsc recognition and prion replication, a number of different epitope tagged mouse PrPc molecules were created in which these particular regions of the molecules were either deleted, scrambled or mutated. For example, within PrP sequences 23-33 and 89-112, mutants were generated in which positively charged residues where changed to alanine. Further, within PrP region 136-158, multiple mutants were prepared in which the native sequence within segments 136-140 and 149-158 was altered. Each of these novel PrP molecules were individually expressed in scrapie-prion infected N2a cells following transfection. Conversion of the various mutated mouse PrPc substrates to PrPsc was measured and compared quantitatively and qualitatively to the conversion of equivalently expressed epitoipe tagged wild-type mouse PrPc. The results of these experiments will be presented.
AD L. Solforosi, A. Bellon, G. Abalos, J. Cruite, R.A. Williamson, The Scripps Research Insitute, USA
SP englisch
PO Schottland