Prion protein interaction with glycosaminoglycan occurs with the formation of oligomeric complexes stabilized by Cu(II) bridges.
Several lines of evidence have shown glycosaminoglycans (GAGs) to be physiological ligands of the prion protein (PrP), but the molecular and regulatory aspects of the interaction remain unknown. Using full-length recombinant prion protein and low molecular mass heparin and heparan sulfate as glycosaminoglycans, we have found that the interaction occurs with the formation of oligomeric complexes. Within the protein-glycosaminoglycan complexes, PrP exhibited an enhanced fluorescence emission and a reduced solvent exposure. The pH and ionic strength-dependence of the interaction reveals His residues as the main binding sites at acid pH. A synthetic peptide consisting of four octarepeats is able to reproduce the His-dependent binding of the protein, thus demonstrating the role of the octarepeats in the GAG interaction. Alternatively, PrP can bind GAGs through His-bound Cu(II). These Cu(II) bridges promote a tighter interaction, as shown by the increased resistance to ionic strength, to protease action, and to pH-induced cation release. Inspection of other cations shows that Zn(II) but not Ni(II) shares the interaction trend. Taken together, our data suggest that the octarepeat region constitutes a novel GAG-binding sequence and that His-bound Cu(II) may act as a cofactor for intermolecular recognition reactions, allowing the formation of PrP-Cu(II)-glycosaminoglycan assemblies that may be crucial entities in the PrP metabolism.
González-Iglesias
Reinerio
R
Insto Química-Física Rocasolano, CSIC, Serrano 119, 28006 Madrid, Spain.
Pajares
María A
MA
Ocal
Carmen
C
Espinosa
Juan Carlos
JC
Oesch
Bruno
B
Gasset
María
M
D000818:Animals; D002413:Cations, Divalent / Q000378:metabolism / Q000494:pharmacology; D002417:Cattle; D003300:Copper / Q000378:metabolism* / Q000494:pharmacology; D019286:Endopeptidase K / Q000378:metabolism; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism*; D006497:Heparan Sulfate / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008027:Light; D018625:Microscopy, Atomic Force; D009994:Osmolar Concentration; D011328:Prions / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D011485:Protein Binding / Q000187:drug effects; D023302:Protein Footprinting; D020836:Protein Structure, Quaternary / Q000187:drug effects; D012542:Scattering, Radiation; D012995:Solubility; D012997:Solvents; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared
D002413:Cations, Divalent; D003300:Copper; D019286:Endopeptidase K; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D008024:Ligands; D011328:Prions; D012997:Solvents