NR AXSB
AU Mangels,C.; Leliveld,R.S.; Müller-Schiffmann,A.; Schweimer,K.; Ziegler,J.; Petsch,B.; Stitz,L.; Rösch,P.; Korth,C.; Schwarzinger,S.
TI NMR and Fluorescence Structural Studies of an Anti-Prion Protein scFv Fragment
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Oral Abstracts FC1.1
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Vortrag
AB
Background: The conversion of cellular prion protein, PrPc, into its scrapie isoform PrPsc is a key event in prion diseases. Preventing this structural transition is a promising route to therapy. We have chosen PrPc as a target for intervention by means of antibody fragments. In particular, helix 1 of PrP (PrPc-H1) has previously been shown to be a suitable target for a potential antibody-therapy (Korth, 1997). Single chain variable fragments (scFv) from a hybridoma cell line W226 generated through immunization of PrP ko mice with purified PrPsc were shown to bind to PrPc with subnanomolar affinity and to exhibit strong antiprion activity.
Aims: We aim to structurally characterize this novel scFvW226 in its complex with PrPc in order to understand the molecular basis of prion propagation inhibition by helix 1 targeting antibodies.
Methods: The antibody epitope has been mapped by fluorescence spectroscopy using alanine-scanning. The solution structure of the isolated antibody fragment as well as of its complex with PrPc-H1 is studied by means of high-field TROSY-NMRspectroscopy.
Results: Our fluorescence studies revealed that three residues in PrPc-H1 make essential contributions to binding. Surprisingly, the residues involved in this highaffinity interaction form a very polar, charged binding surface for scFvW226. NMR studies of the free scFvW226 and of the bound form with PrPc-H1 show clear differences in the spectra allowing to allocate the binding region on the antibody part.
Discussion: The structural description of the binding interface of this therapeutically active antibody-fragment with its epitope provides first insight into the underlying principles of the high-affinity binding to PrP. Ultimately, knowledge of the high resolution structure of the complex of these two proteins builds the foundation for the rational design of novel anti-prion drugs.
AD C. Mangels, K. Schweimer, J. Ziegler, P. Rösch, S. Schwarzinger, University of Bayreuth, Department of Biopolymers, Germany; R. S. Leliveld, A. Müller-Schiffmann, C. Korth, Heinrich-Heine-University, Institute of Neuropathology, Germany; B. Petsch, L. Stitz, Friedrich-Löffler Institut, Institute of Immunology, Germany
SP englisch
PO Schottland