Biophys J, July 2001, p. 516-525, Vol. 81, No. 1

Electron Paramagnetic Resonance Evidence for Binding of Cu²⁺ to the C-terminal Domain of the Murine Prion Protein

Grazia M. Cereghetti,^* Arthur Schweiger, Rudi Glockshuber,^* and Sabine Van Doorslaer

 ^*Institute of Molecular Biology and Biophysics, Swiss Federal Institute of Technology, Hönggerberg, CH-8093 Zurich, Switzerland and  Laboratory for Physical Chemistry, Swiss Federal Institute of Technology, ETH Zentrum, CH-8092 Zurich, Switzerland

Transmissible spongiform encephalopathies in mammals are believed to be caused by scrapie form of prion protein (PrP^Sc), an abnormal, oligomeric isoform of the monomeric cellular prion protein (PrP^C). One of the proposed functions of PrP^C in vivo is a Cu(II) binding activity. Previous studies revealed that Cu²⁺ binds to the unstructured N-terminal PrP^C segment (residues 23-120) through conserved histidine residues. Here we analyzed the Cu(II) binding properties of full-length murine PrP^C (mPrP), of its isolated C-terminal domain mPrP(121-231) and of the N-terminal fragment mPrP(58-91) in the range of pH 3-8 with electron paramagnetic resonance spectroscopy. We find that the C-terminal domain, both in its isolated form and in the context of the full-length protein, is capable of interacting with Cu²⁺. Three Cu(II) coordination types are observed for the C-terminal domain. The N-terminal segment mPrP(58-91) binds Cu²⁺ only at pH values above 5.0, whereas both mPrP(121-231) and mPrP(23-231) already show identical Cu(II) coordination in the pH range 3-5. As the Cu²⁺-binding N-terminal segment 58-91 is not required for prion propagation, our results open the possibility that Cu²⁺ ions bound to the C-terminal domain are involved in the replication of prions, and provide the basis for further analytical studies on the specificity of Cu(II) binding by PrP.

Biophys J, July 2001, p. 516-525, Vol. 81, No. 1
© 2001 by the Biophysical Society   0006-3495/01/07/516/10  $2.00

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