title: Identification of interaction domains of the prion protein with its 37-kDa/67-kDa laminin receptor.
abstract: 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.
authors.affiliation: Laboratorium für Molekulare Biologie-Genzentrum-Institut für Biochemie der LMU München, Feodor-Lynen Strasse 25, D-81377 Munich, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Hundt
authors.forename: J M
authors.initials: JM
authors.lastname: Peyrin
authors.forename: S
authors.initials: S
authors.lastname: Haïk
authors.forename: S
authors.initials: S
authors.lastname: Gauczynski
authors.forename: C
authors.initials: C
authors.lastname: Leucht
authors.forename: R
authors.initials: R
authors.lastname: Rieger
authors.forename: M L
authors.initials: ML
authors.lastname: Riley
authors.forename: J P
authors.initials: JP
authors.lastname: Deslys
authors.forename: D
authors.initials: D
authors.lastname: Dormont
authors.forename: C I
authors.initials: CI
authors.lastname: Lasmézas
authors.forename: S
authors.initials: S
authors.lastname: Weiss
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000502:physiology; D016466:CHO Cells; D002460:Cell Line; D002850:Chromatography, Gel; D006224:Cricetinae; D005697:Galactosides / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics; D019812:Heparan Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009842:Oligopeptides; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000235:genetics; D011328:Prions / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000502:physiology; D017474:Receptors, Laminin / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012672:Semliki forest virus / Q000235:genetics; D020798:Two-Hybrid System Techniques
chemical_list: D005697:Galactosides; D019812:Heparan Sulfate Proteoglycans; D009842:Oligopeptides; D010446:Peptide Fragments; D010455:Peptides; D011328:Prions; D011498:Protein Precursors; D017474:Receptors, Laminin; D011993:Recombinant Fusion Proteins; C007816:beta-galactoside; C085706:FLAG peptide; D005982:Glutathione Transferase