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  • The EMBO Journal (2001) 20, 5876 - 5886
  • doi:10.1093/emboj/20.21.5876

Identification of interaction domains of the prion protein with its 37-kDa/67-kDa laminin receptor

Christoph Hundt1,3, Jean-Michel Peyrin2,3, Stéphane Haïk2,3, Sabine Gauczynski1, Christoph Leucht1, Roman Rieger1, Maria Louise Riley1, Jean-Philippe Deslys2, Dominique Dormont2, Corinne Ida Lasmézas2,4 and Stefan Weiss1,4

  1. Laboratorium für Molekulare Biologie-Genzentrum-Institut für Biochemie der LMU München, Feodor-Lynen Strasse 25, D-81377 Munich, Germany
  2. CEA, Service de Neurovirologie, DRM/DSV, CRSSA, 18, Route du Panorama, BP.6, F-92265 Fontenay-aux-Roses Cedex, France
  3. C.Hundt, J.-M.Peyrin and S.Haïk contributed equally to this work
  4. C.I.Lasmézas and S.Weiss should be considered as the senior authors of this work

Correspondence to:

Corinne Ida Lasmézas, E-mail: corinne.lasmezas@cea.fr

Stefan Weiss, E-mail: Weiss@lmb.uni-muenchen.de

Received 1 December 2000; Accepted 5 September 2001; Revised 9 August 2001

Cell-binding and internalization studies on neuronal and non-neuronal cells have demonstrated that the 37-kDa/67-kDa laminin receptor (LRP/LR) acts as the receptor for the cellular prion protein (PrP). Here we identify direct and heparan sulfate proteoglycan (HSPG)-dependent interaction sites mediating the binding of the cellular PrP to its receptor, which we demonstrated in vitro on recombinant proteins. Mapping analyses in the yeast two-hybrid system and cell-binding assays identified PrPLRPbd1 [amino acids (aa) 144–179] as a direct and PrPLRPbd2 (aa 53–93) as an indirect HSPG-dependent laminin receptor precursor (LRP)-binding site on PrP. The yeast two-hybrid system localized the direct PrP-binding domain on LRP between aa 161 and 179. Expression of an LRP mutant lacking the direct PrP-binding domain in wild-type and mutant HSPG-deficient Chinese hamster ovary cells by the Semliki Forest virus system demonstrates a second HSPG-dependent PrP-binding site on LRP. Considering the absence of LRP homodimerization and the direct and indirect LRP–PrP interaction sites, we propose a comprehensive model for the LRP–PrP–HSPG complex.

  • Keywords:

    • heparan sulfate proteoglycans,
    • in vitro interaction,
    • 37-kDa laminin receptor precursor,
    • 67-kDa high-affinity laminin receptor,
    • LRP-PrP-binding domains