NR AFRO
AU Hundt,C.; Peyrin,J.M.; Haik,S.; Gauczynski,S.; Leucht,C.; Rieger,R.; Riley,M.L.; Deslys,J.P.; Dormont,D.; Lasmezas,C.I.; Weiss,S.
TI Identification of interaction domains of the prion protein with its 37-kDa/67-kDa laminin receptor
QU EMBO Journal 2001 Nov 1; 20(21): 5876-86
PT journal article
AB 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.
MH Animal; Binding Sites/physiology; CHO Cells; Cell Line; Chromatography, Gel; Galactosides/metabolism; Glutathione Transferase/genetics; Hamsters; Heparan Sulfate Proteoglycan/genetics/*metabolism; Human; Mice; Peptide Fragments/genetics/metabolism; Peptides/genetics; Prions/genetics/*metabolism; Protein Binding/physiology; Protein Precursors/genetics/*metabolism; Protein Structure, Tertiary/physiology; Receptors, Laminin/genetics/*metabolism; Recombinant Fusion Proteins/genetics/metabolism; Semliki forest virus/genetics; Support, Non-U.S. Gov't; *Two-Hybrid System Techniques
AD Laboratorium für Molekulare Biologie-Genzentrum-Institut für Biochemie der LMU München, Feodor-Lynen Strasse 25, D-81377 Munich, Germany.
SP englisch
PO England