Identification of the heparan sulfate binding sites in the cellular prion protein.
Data from cell culture and animal models of prion disease support the separate involvement of both heparan sulfate proteoglycans and copper (II) ions in prion (PrP) metabolism. Though direct interactions between prion protein and heparin have been recorded, little is known of the structural features implicit in this interaction or of the involvement of copper (II) ions. Using biosensor and enzyme-linked immunosorbent assay methodology we report direct heparin and heparan sulfate-binding activity in recombinant cellular prion protein (PrP(c)). We also demonstrate that the interaction of recombinant PrP(c) with heparin is weakened in the presence of Cu(II) ions and is particularly sensitive to competition with dextran sulfate. Competitive inhibition experiments with chemically modified heparins also indicate that 2-O-sulfate groups (but not 6-O-sulfate groups) are essential for heparin recognition. We have also identified three regions of the prion protein capable of independent binding to heparin and heparan sulfate: residues 23-52, 53-93, and 110-128. Interestingly, the interaction of an octapeptide-spanning peptide motif amino acids 53-93 with heparin is enhanced by Cu(II) ions. Significantly, a peptide of this sequence is able to inhibit the binding of full-length prion molecule to heparin, suggesting a direct role in heparin recognition within the intact protein. The collective data suggest a complex interaction between prion protein and heparin/heparan sulfate and has implications for the cellular and pathological functions of prion proteins.
Warner
Richard G
RG
Molecular Cell Biology Laboratories, School of Biosciences, University of Birmingham, Edgbaston, Birmingham, England B15 2TT, United Kingdom.
Hundt
Christoph
C
Weiss
Stefan
S
Turnbull
Jeremy E
JE
D000818:Animals; D001665:Binding Sites; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D005982:Glutathione Transferase / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism*; D018621:PrPC Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D005982:Glutathione Transferase; D006493:Heparin; D006497:Heparan Sulfate; D018621:PrPC Proteins; D011993:Recombinant Fusion Proteins