NR AXSC
AU Mangels,C.; Ziegler,J.; Pischel,S.; Wöhrl,B.M.; Rösch,P.; Schwarzinger,S.
TI NMR Structural Studies of the Prion Protein - Laminin Receptor Precursor Complex
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.37
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: The pathogenic conversion of cellular prion protein, PrPc, into its scrapie isoform, PrPsc, may be influenced by other proteins. The 37/67 kDa laminin receptor precursor (LRP) has been shown to bind to extracellular PrP (Gauczynski 2001) and to be involved in prion propagation (Leucht 2003). Knowledge of the details of the molecular interactions of these two proteins is therefore of high interest. The binding domains of both proteins have previously been coarsely determined by yeast-2-hybrid (Y2H) assays (Hundt 2001).
Aims: We aim to structurally characterize the binding surface of PrPc with LRP.
Methods: The detailed structural properties of the PrP-LRP interaction surface are elucidated by means of solution NMR-spectroscopy. In particular, we apply a combination of paramagnetic relaxation enhancement (PRE) and chemical shift perturbation (CSP) to identify residues responsible for binding.
Results: We show that the peptides corresponding to the isolated binding domains of PrP and LRP do not interact in solution. However, weak binding could be detected using spin-labelled peptides mimicking the binding domain of LRP and full length PrP by measuring PRE. These data indicate binding in a region similar, but not identical to the binding site suggested from Y2H-assays. In particular, binding takes place on the surface defined by helices 1 and 3 of PrP. CSP studies of full length PrP and the extracellular domain of LRP confirm these findings.
Discussion: Due to the size of the complex of both proteins (~47 kDa) we started from isolated binding domains, which did not interact. Presumably, this is due to the lack of stable conformers responsible for binding. Conformational stabilization of the isolated peptide fragments by including them into their native tertiary context lead to binding. The difference to the binding site found by Y2H most likely is due to the use of peptide fragments only, which may lead to substantial loss of conformational and tertiary information. Knowledge of such binding interfaces at high resolution builds the foundation for the rational design of novel anti-prion drugs.
AD C. Mangels, J. Ziegler, S. Pischel, B.M. Wöhrl, P. Rösch, S. Schwarzinger, Universtity of Bayreuth, Department of Biopolymers, Germany
SP englisch
PO Schottland