Location and properties of metal-binding sites on the human prion protein.
Although a functional role in copper binding has been suggested for the prion protein, evidence for binding at affinities characteristic of authentic metal-binding proteins has been lacking. By presentation of copper(II) ions in the presence of the weak chelator glycine, we have now characterized two high-affinity binding sites for divalent transition metals within the human prion protein. One is in the N-terminal octapeptide-repeat segment and has a K(d) for copper(II) of 10(-14) M, with other metals (Ni(2+), Zn(2+), and Mn(2+)) binding three or more orders of magnitude more weakly. However, NMR and fluorescence data reveal a previously unreported second site around histidines 96 and 111, a region of the molecule known to be crucial for prion propagation. The K(d) for copper(II) at this site is 4 x 10(-14) M, whereas nickel(II), zinc(II), and manganese(II) bind 6, 7, and 10 orders of magnitude more weakly, respectively, regardless of whether the protein is in its oxidized alpha-helical (alpha-PrP) or reduced beta-sheet (beta-PrP) conformation. A role for prion protein (PrP) in copper metabolism or transport seems likely and disturbance of this function may be involved in prion-related neurotoxicity.
Jackson
G S
GS
Medical Research Council Prion Unit, Department of Neurogenetics, Imperial College School of Medicine at St. Mary's, London W2 1NY, United Kingdom.
Murray
I
I
Hosszu
L L
LL
Gibbs
N
N
Waltho
J P
JP
Clarke
A R
AR
Collinge
J
J
D000595:Amino Acid Sequence; D001665:Binding Sites; D002413:Cations, Divalent / Q000737:chemistry; D003300:Copper / Q000737:chemistry; D005998:Glycine / Q000737:chemistry; D006801:Humans; D008345:Manganese / Q000737:chemistry; D008670:Metals / Q000737:chemistry*; D008969:Molecular Sequence Data; D009532:Nickel / Q000737:chemistry; D011328:Prions / Q000737:chemistry*; D015032:Zinc / Q000737:chemistry
D002413:Cations, Divalent; D008670:Metals; D011328:Prions; D008345:Manganese; D003300:Copper; D009532:Nickel; D015032:Zinc; D005998:Glycine