Prion diseases are caused by the misfolding and aggregation of the prion protein (PrP). Herein we provide evidence that the CuII adduct of the unstructured amyloidogenic fragment of the human PrP (PrP(91-126)) is redox active under physiological conditions. We have identified that the relevant high-affinity CuII binding region of PrP(91-126) is contained between residues 106 and 114. Both [CuII(PrP(91-126))] and [CuII(PrP(106-114))] have CuII Kd values of approximately 90 microM. Furthermore, the smaller PrP fragment PrP(106-114) coordinates CuII producing an electronic absorption spectrum nearly identical with [CuII(PrP(91-126))] (lambda max approximately 610 nm (epsilon approximately 125 M-1 cm-1)) suggesting a similar coordination environment for CuII. Cu K-edge X-ray absorption spectroscopy (XAS) reveals a nearly identical CuN(N/O)2S coordination environment for these two metallopeptides (2N/O at approximately 1.97 A; 1S at approximately 2.30 A; 1 imidazole N at approximately 1.95 A). Both display quasireversible CuII/CuI redox couples at approximately -350 mV vs Ag/AgCl. ESI-MS indicates that both peptides will coordinate CuI. However, XAS indicates differential coordination environments between [CuI(PrP(91-126))] and [CuI(PrP(106-114))]. These data indicate that [CuI(PrP(91-126))] contains Cu in a four coordinate (N/O)2S2 environment with similar (N/O)-Cu bond distances (Cu-(N/O) r = 2.048(4) A), while [CuI(PrP(106-114))] contains Cu in a four coordinate (N/O)2S2 environment with differential (N/O)-Cu bond distances (Cu-(N/O) r1 = 2.057(6) A; r2 = 2.159(3) A). Despite the differential coordination environments both Cu-metallopeptides will catalytically reduce O2 to O2*- at comparable rates.