Key points are not available for this paper at this time.
The prion protein (PrP) is a Cu2+ binding cell surface glycoprotein that can misfold into a β-sheet-rich conformation to cause prion diseases. The majority of copper binding studies have concentrated on the octarepeat region of PrP. However, using a range of spectroscopic techniques, we show that copper binds preferentially to an unstructured region of PrP between residues 90 and 115, outside of the octarepeat domain. Comparison of recombinant PrP with PrP-(91–115) indicates that this prion fragment is a good model for Cu2+ binding to the full-length protein. In contrast to previous reports we show that Cu2+ binds to this region of PrP with a nanomolar dissociation constant. NMR and EPR spectroscopy indicate a square-planar or square-pyramidal Cu2+ coordination utilizing histidine residues. Studies with PrP analogues show that the high affinity site requires both His96 and His111 as Cu2+ ligands, rather than a complex centered on His96 as has been previously suggested. Our circular dichroism studies indicate a loss of irregular structure on copper coordination with an increase in β-sheet conformation. It has been shown that this unstructured region, between residues 90 and 120, is vital for prion propagation and different strains of prion disease have been linked with copper binding. The role of Cu2+ in prion misfolding and disease must now be re-evaluated in the light of these findings. The prion protein (PrP) is a Cu2+ binding cell surface glycoprotein that can misfold into a β-sheet-rich conformation to cause prion diseases. The majority of copper binding studies have concentrated on the octarepeat region of PrP. However, using a range of spectroscopic techniques, we show that copper binds preferentially to an unstructured region of PrP between residues 90 and 115, outside of the octarepeat domain. Comparison of recombinant PrP with PrP-(91–115) indicates that this prion fragment is a good model for Cu2+ binding to the full-length protein. In contrast to previous reports we show that Cu2+ binds to this region of PrP with a nanomolar dissociation constant. NMR and EPR spectroscopy indicate a square-planar or square-pyramidal Cu2+ coordination utilizing histidine residues. Studies with PrP analogues show that the high affinity site requires both His96 and His111 as Cu2+ ligands, rather than a complex centered on His96 as has been previously suggested. Our circular dichroism studies indicate a loss of irregular structure on copper coordination with an increase in β-sheet conformation. It has been shown that this unstructured region, between residues 90 and 120, is vital for prion propagation and different strains of prion disease have been linked with copper binding. The role of Cu2+ in prion misfolding and disease must now be re-evaluated in the light of these findings. Prion diseases are fatal neurodegenerative diseases that include Creutzfeldt-Jacob disease in humans, mad cow disease in cattle, and scrapie in sheep. The infectious agent in transmissible spongiform encephalopathies is a proteinous infectious particle or “prion,” which is devoid of nucleic acid. It is believed that these spongiform encephalopathies are caused by the accumulation of an abnormally folded isoform of the cellular prion protein (PrPC). 1The abbreviations used are: PrPC, cellular isoform of prion protein; PrP, prion protein; PrPSc, scrapie isoform of prion protein; TOCSY, total correlation spectroscopy; NOESY, nuclear Overhauser exchange spectroscopy. This misfolded protein is rich in β-sheet and is designated the scrapie isoform, PrPSc (1Prusiner S.B. Science. 1997; 278: 245-251Google Scholar, 2Horwich A.L. Weissman J.S. Cell. 1997; 89: 499-510Google Scholar, 3Prusiner S.B. Proc. Natl. Acad. Sci. U. S. A. 1998; 95: 13363-13383Google Scholar). Whereas normal physiological function of the prion protein is yet to be determined, the ability of PrPC to bind Cu2+in vivo and in vitro suggests a role in copper homeostasis (4Brown D.R. Qin K. Herms J.W. Madlung A. Manson J. Strome R. Fraser P.E. Kruck T. von Bohlen A. Schulz-Schaeffer W. Giese A. Westaway D. Kretzschmar H. Nature. 1997; 390: 684-687Google Scholar, 5Viles J.H. Cohen F.E. Prusiner S.B. Goodin D.B. Wright P.E. Dyson J.H. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 2042-2047Google Scholar). Indeed, elevated copper levels promote endocytosis of PrPC suggesting that PrP could transport copper into the cell (6Pauly P.C. Harris D.A. J. Biol. Chem. 1998; 273: 33107-33110Google Scholar, 7Perera W.S. Hooper N.M. Curr. Biol. 2001; 11: 519-523Google Scholar). Recent data has shown that PrPC expression increased the binding of copper to the outer plasma cell membrane and increases antioxidant enzyme activities (8Rachidi W. Vilette D. Guiraud P. Arlotto M. Riondel J. Laude H. Lehmann S. Favier A. J. Biol. Chem. 2003; 278: 9064-9072Google Scholar). Furthermore, it is suggested that PrPC plays a protective role by binding Cu2+ in a redox inactive state (9Wong B.S. Pan T. Liu T. Li R. Petersen P. D.R. Scholar, M. Scholar). role for is as it D.R. B.S. J. 1999; Scholar, B.S. Pan T. Liu T. Li R. P. 273: Scholar, D.R. A. J. 1998; Scholar, T. M. B.S. Li R. J. D.R. J. 2003; Scholar). The cellular prion protein is a cell surface glycoprotein a the are now a of NMR of in the R. S. M. R. K. Nature. Scholar, Liu H. S. H. K. D. Prusiner S.B. Cohen F.E. Proc. Natl. Acad. Sci. U. S. A. 1997; Scholar, J.H. D. Cohen F.E. Prusiner S.B. Wright P.E. Dyson J.H. Proc. Natl. Acad. Sci. U. S. A. 1997; Scholar, R. Liu A. T. R. von M. K. Proc. Natl. Acad. Sci. U. S. A. Scholar). structure of PrPC has been The structure is between M. W. M. Biol. 2001; Scholar). PrPC In the of the residues is unstructured J.H. D. Cohen F.E. Prusiner S.B. Wright P.E. Dyson J.H. Proc. Natl. Acad. Sci. U. S. A. 1997; and has a high of in the of copper J.H. D. Prusiner S.B. Cohen F.E. Dyson Wright P.E. 2001; Scholar). In the residues is R. S. M. R. K. Nature. Scholar). of an which is It is this unstructured region that binds Cu2+ J.H. Cohen F.E. Prusiner S.B. Goodin D.B. Wright P.E. Dyson J.H. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 2042-2047Google Scholar). binding of a Cu2+ to PrP has been (4Brown D.R. Qin K. Herms J.W. Madlung A. Manson J. Strome R. Fraser P.E. Kruck T. von Bohlen A. Schulz-Schaeffer W. Giese A. Westaway D. Kretzschmar H. Nature. 1997; 390: 684-687Google Scholar, A. S. S. Kretzschmar H. J. Biol. Chem. 2001; Scholar, J. Proc. Natl. Acad. Sci. U. S. A. 2001; as as Cu2+ coordination to a fragment of PrP D.R. J. Scholar). The majority of studies have concentrated on the copper binding to the octarepeat region of PrP. Recent on copper binding to the include circular dichroism studies J.H. J. Biol. Chem. 2003; 278: spectroscopy studies J. Cohen F.E. Prusiner S.B. and a P. A. J. Scholar). for Cu2+ binding to the octarepeat region have been to be between and A. S. S. Kretzschmar H. J. Biol. Chem. 2001; Scholar, J.H. J. Biol. Chem. 2003; 278: have suggested a affinity with a in the range J. Proc. Natl. Acad. Sci. U. S. A. 2001; Scholar). Studies with have shown that a of PrP with the octarepeat region are to prion D. M. A. von A. Scholar). This has that a between copper and prion disease to be However, studies copper can bind to the unstructured of PrP outside of the octarepeat region with a dissociation J. Proc. Natl. Acad. Sci. U. S. A. 2001; Scholar). His111 His96 have been in this binding J. Proc. Natl. Acad. Sci. U. S. A. 2001; Scholar, J. J. Biol. 2001; Scholar, Prusiner S.B. J. 2003; Scholar). the unstructured region between the and the is to be for and in prion disease T. M. Cohen F.E. Prusiner S.B. Proc. Natl. Acad. Sci. U. S. A. Scholar, M. T. A. A. M. S. A. J. Scholar, T. M. Cohen F.E. Prusiner S.B. 1997; Scholar, D.R. Kretzschmar Nature. Scholar). in this unstructured region are to a role in the misfolding of PrPC into the β-sheet-rich conformation R. P. M. Proc. Natl. Acad. Sci. U. S. A. Scholar). The of a high affinity Cu2+ site the region have for the copper structure in this region of can the cellular prion protein into a R. Harris D.A. J. Biol. Chem. 2001; Scholar). in this region the different of PrP with and the of Cu2+ K. Kretzschmar Fraser P.E. Westaway D. J. Biol. Chem. these to in different strains of prion disease S. J. Biol. 1999; Scholar). In a fragment this unstructured region, a of residues and has been shown to be in both the and D.R. Kretzschmar Nature. Scholar, T. D. D.R. A. A. J. 2003; Scholar). binding to this has been linked with M. R. 2001; Scholar). are a of prion disease R. R. D.R. J. and binding to the prion protein is in prion disease B.S. M. Pan T. Liu T. Li R. P. D.R. J. 2001; Scholar). it has been suggested that a loss of PrP function in copper transport and homeostasis be a of prion Recent studies have shown that scrapie copper a cellular W. A. A. Guiraud P. Riondel J. M. Favier A. Lehmann S. J. Biol. Chem. 2003; 278: and that redox of PrP is in prion disease D. Prusiner S.B. Proc. Natl. Acad. Sci. U. S. A. 2001; Scholar, Scholar). of the copper binding octarepeat region has been shown to disease D. M. A. von A. Scholar). a that disease in is with the of copper D.R. H. R. T. J. Biol. Chem. 2003; 278: Scholar). of PrP with prion disease and to endocytosis W.S. Hooper N.M. Curr. Biol. 2001; 11: 519-523Google Scholar). The role of copper in the normal function of PrP, as as in prion has been the of a of D.R. 2001; Scholar, D.R. 2001; Scholar, S. Curr. Chem. Biol. Scholar, Herms J. J. 2003; Scholar, Chem. Scholar). The of this is to the binding of Cu2+ to the unstructured region of PrP outside of the This region is of it is for and in are in the that we to the of this Cu2+ binding site are with an EPR suggesting His96 as the for Cu2+ coordination Prusiner S.B. J. 2003; an suggested and His96 as J. J. Biol. 2001; Scholar). In the for Cu2+ in the between the (4Brown D.R. Qin K. Herms J.W. Madlung A. Manson J. Strome R. Fraser P.E. Kruck T. von Bohlen A. Schulz-Schaeffer W. Giese A. Westaway D. Kretzschmar H. Nature. 1997; 390: 684-687Google Scholar, A. S. S. Kretzschmar H. J. Biol. Chem. 2001; and J. Proc. Natl. Acad. Sci. U. S. A. 2001; dissociation with the octarepeat PrP-(91–115) and and PrP-(91–115) analogues and have been used to residues in Cu2+ spectroscopic and EPR to the coordination and of copper binding to the prion protein. and of Prion region of the full-length with a region, into a to a protein as previously D.R. B.S. J. 1999; Scholar). The protein in of to an of and the protein expression by the of to and in a and The of by for The protein to a affinity The protein the using The of the protein by and The protein by of in concentrated using and with a to The of by and of PrP the and the using high and using the the full-length prion the with and the and NMR spectroscopy the of (PrP) and in the of or the using of or The using of by the of used for von Scholar). The by The of or using affinity of for Cu2+ is the is or a dissociation of and or The dissociation is for Scholar). on an a cell with a used for between and with cell used for data between and with a of and the or using and the The in to using the is the and the The is in of M. with a using a NMR on a using a in or or for the Cu2+ and K. In the using the M. H. J. 1998; in with a in the using the total correlation spectroscopy with complex with using the The a for with a of A. J. Scholar). Overhauser exchange spectroscopy J. P. J. Chem. with data with of and to by to both the to and both to using on a and using the M. M. K. J. Scholar). and for PrP-(91–115) using the and data and K. NMR of and Scholar). of of the and of histidine EPR on a a of The a of a of a of and a of or K. The of the to the are to into of PrP-(91–115) in the region are shown in In the of PrP-(91–115) an in with the structure of full-length PrP in this region J.H. D. Cohen F.E. Prusiner S.B. Wright P.E. Dyson J.H. Proc. Natl. Acad. Sci. U. S. A. 1997; Scholar). that of Cu2+ increases the centered with a in the is with Cu2+ a of binding to of The in the with Cu2+ are by the shown as an The in the copper coordination indicate an increase in β-sheet or by a loss of irregular with Cu2+ and binding to His111 and His96 residues are as and cause the to on β-sheet in this region, the shown in this show that Cu2+ to both His111 and The data indicates that the a irregular a β-sheet between His111 and His96 is and we have the of Cu2+ binding to the prion protein PrP-(91–115) with of Cu2+ and the a of between and the a is to Cu2+ the increases the increases in is a in the to suggesting a in the coordination of the Cu2+ The and and for the complex are for a Cu2+ The can be to using model of copper the for Cu2+ with a to for Cu2+ with J. Biol. Chem. Scholar). The are of or in a square-planar dichroism of of Cu2+ have been and shown in data show that the of the Cu2+ with The a and In a is with The is a of the the and of the and good with the using a of and of of Cu2+ in the as shown in coordination to PrP-(91–115) is to The of histidine a for the of binding. The of the indicate a the this coordination the or J. Chem. Scholar). The in coordination and as the is the of histidine is to be of the coordination of as the is H. R. Chem. Scholar). the copper binding of of Cu2+ to and the and The of Cu2+ binding to PrP-(91–115) is shown in The increases of Cu2+ have been the a of the and a increase in of the The the of Cu2+ as a function of the and is a in the binding of Cu2+ that the Cu2+ PrP The of Cu2+ indicates that is a in the affinity between the of the and Cu2+ is a in the binding of this is the Cu2+ In the of Cu2+ is The using circular dichroism and and the between and the binding the are shown in and the binding in D. The binding show that the and have of Cu2+ is of Cu2+ is a in the In with the it is that Cu2+ binds to PrP is a in the binding of the to increase in of suggesting Cu2+ binding to PrP. the and between and of suggesting that is a binding site that Cu2+ is It is the data in and that binding of Cu2+ to PrP-(91–115) or of as of Cu2+ in increases in and are of Cu2+ in as copper in is and a which is In an to Cu2+ binding to PrP-(91–115) the affinity Cu2+ binding the prion fragment has been in the of is a and a in the region In the of the of PrP-(91–115) with of to the of PrP of copper is This suggests that copper binds to PrP with a affinity than as of Cu2+ are in the of the to a copper to The affinity of Cu2+ binding to PrP-(91–115) is to physiological the affinity of Cu2+ binding to PrP-(91–115) we have used as a copper can bind to Cu2+ with of both the and for Scholar). The are the region for PrP-(91–115) as the is to of The a of of with and to with PrP-(91–115) for Cu2+ of be to both Cu2+ the that the of a copper has been of to has been a copper has been Furthermore, of a of copper has been the affinity copper site with of This that the affinity site has a that of of the the as is with of It that the affinity site binds to copper an of in affinity than the In a used to for Cu2+ the of of to has a for Cu2+ of and for Scholar). is and a shown in The of is PrP a in the and an increase in the However, the of has been to bind of the Cu2+ to the high affinity site This is the in The the of in the of of with a that is a of and the are histidine has a and of histidine to Cu2+ the and of to Cu2+ the these studies it is that PrP-(91–115) bind than of The of Cu2+ bind to PrP with an affinity with the affinity binding site for Cu2+ in the of or Cu2+ binds to PrP-(91–115) with an affinity than to a nanomolar dissociation for the has been in copper binding to the octarepeat region of PrP. have the of the octarepeat on Cu2+ binding to the PrP-(91–115) as the Cu2+ of PrP-(91–115) of between copper binding to the of PrP. the of the of to The and J.H. Cohen F.E. Prusiner S.B. Goodin D.B. Wright P.E. Dyson J.H. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 2042-2047Google Scholar, J.H. J. Biol. Chem. 2003; 278: are with the of of to copper PrP-(91–115) to bind the Cu2+ to the octarepeat in a The is to the shown in for the of PrP-(91–115) is used the have the of In this of the copper is by This is the is as shown in In this an of is and of of the copper the affinity site of The in a of on a of have shown that for Cu2+ to J.H. J. Biol. Chem. 2003; 278: Scholar). This is in with this which that copper bind to PrP-(91–115) with a or a affinity than that a Cu2+ preferentially bind to PrP-(91–115) rather than the octarepeat region we have and data for the PrP residues In we data for is and a in which the have been the of the protein between and This residues for prion propagation with residues shown in indicate that PrP-(91–115) is a good model for Cu2+ binding to full-length PrP. of of copper binding to The show a to the range is which and to The is for with a and of the is in the for between and of the is between and the of on Cu2+ binding to with can the of Cu2+ that is as with the PrP-(91–115) the of of to the total of Cu2+ the high affinity Cu2+ binding site is by in we as with the Cu2+ the high affinity Cu2+ binding site indicates a Cu2+ preferentially bind to the region between residues and 115, rather than the of PrP. The PrP-(91–115) fragment in an to It is this data in that the PrP-(91–115) fragment a model for copper binding to full-length PrP physiological full-length PrP is unstructured between residues and and as a the PrP fragment is a good model for Cu2+ binding. is to coordination the copper this the of show good with In as be the show that the PrP-(91–115) has the affinity for Cu2+ as EPR used to the of the the copper EPR of PrP-(91–115) and with and of Cu2+ are shown in and In with data the EPR indicate a suggesting a square-planar for both and of Cu2+ and J. have shown that and of the of the of Cu2+ a of are with and of and a the EPR are of coordination of and or and the is a of are physiological these are the is to the of have The and are and the in the are to or to the is a of Cu2+ is an of with and and are This the that a in to of a to of the EPR data for PrP-(91–115) a range of that the binds copper in a coordination and The EPR for PrP-(91–115) of which the of on the and the of coordination a is of the EPR to is between the Cu2+ The and of the is by the to The of in the indicates that histidine between copper as is in is J.S. Proc. Natl. Acad. Sci. U. S. A. Scholar). of Cu2+ on the NMR of PrP-(91–115) to the of the copper binding and used to in the to for NMR that of to histidine and for both His96 and His111 are than Furthermore, His96 and His111 are to an Cu2+ The of The the Cu2+ in of NMR linked to the by or The levels of Cu2+ used the of of the of to the copper binding The NMR that the of both His111 and His96 are in coordination of the on EPR data the of the of is of this in to His111 and the rather than In an to the residues in Cu2+ binding to with His96 and His111 with an and the in the region of and are shown with of Cu2+ and different that for the fragment of of the for and to that PrP-(91–115) that that and of the for the analogues are between and the of PrP-(91–115) as shown in this data is that the high affinity copper site requires the of both His96 and His111 residues to a the affinity of copper for the The His111 by the of a loss of as shown in The binds copper of the copper is that Cu2+ binds with a affinity to that of The indicate that both and have an affinity for Cu2+ than PrP-(91–115) to of that Cu2+ requires both His96 and His111 residues to a that the for and are of for both analogues are centered with the of the The can be used to in of coordination the J. Chem. Scholar). The of the indicates the of the to the Cu2+ coordination and this it is that the of the are This coordination to the residues of His96 and to the of The of is to of a model M. M. M. H. J. J. 2003; Scholar). This that binding of copper to is the of His96 and to the of In we have that the affinity site is a complex that requires both His111 and are in It is now that physiological of the binds a Cu2+ J.H. J. Biol. Chem. 2003; 278: Scholar, J. Cohen F.E. Prusiner S.B. Scholar). In copper has been suggested to bind to a on PrP (4Brown D.R. Qin K. Herms J.W. Madlung A. Manson J. Strome R. Fraser P.E. Kruck T. von Bohlen A. Schulz-Schaeffer W. Giese A. Westaway D. Kretzschmar H. Nature. 1997; 390: 684-687Google Scholar, A. S. S. Kretzschmar H. J. Biol. Chem. 2001; Scholar, Cohen F.E. A.L. Prusiner S.B. Sci. Scholar). His96 His111 have been in copper coordination J. Proc. Natl. Acad. Sci. U. S. A. 2001; Scholar, J. J. Biol. 2001; Scholar, Prusiner S.B. J. 2003; Scholar). have shown that and PrP-(91–115) bind a Cu2+ than the octarepeat studies indicate that residues between and can copper the of copper binds to PrP-(91–115) with a affinity to the The of full-length PrP Cu2+ can bind is spectroscopic studies of full-length PrP in which residues have been has It is that between and binds to Cu2+ in an to the fragment affinity Cu2+ binding to PrP outside of the octarepeat region it is to using a copper affinity have previously for full-length PrP J.H. Cohen F.E. Prusiner S.B. Goodin D.B. Wright P.E. Dyson J.H. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 2042-2047Google Scholar). The with Cu2+ and as a the are by the and the high affinity site EPR studies of full-length PrP the that copper binds to the PrP-(91–115) fragment in an to the full-length protein Prusiner S.B. J. 2003; Scholar). binding in the region has been suggested to have a affinity J. Proc. Natl. Acad. Sci. U. S. A. 2001; Scholar). Our suggests a nanomolar dissociation this is than the dissociation suggested by (4Brown D.R. Qin K. Herms J.W. Madlung A. Manson J. Strome R. Fraser P.E. Kruck T. von Bohlen A. Schulz-Schaeffer W. Giese A. Westaway D. Kretzschmar H. Nature. 1997; 390: 684-687Google Scholar, A. S. S. Kretzschmar H. J. Biol. Chem. 2001; Scholar). The of Cu2+ is in plasma Scholar). It is that a of Cu2+ bind to in plasma S. J. Biol. Chem. Scholar). these it is that PrP has a affinity than for It is that PrP bind Cu2+ physiological The affinity of PrP for copper the in vivo studies that PrP binds Cu2+ (4Brown D.R. Qin K. Herms J.W. Madlung A. Manson J. Strome R. Fraser P.E. Kruck T. von Bohlen A. Schulz-Schaeffer W. Giese A. Westaway D. Kretzschmar H. Nature. 1997; 390: 684-687Google Scholar). on of and analogues we show that of His111 or His96 the coordination that both are to the high affinity Cu2+ In a EPR of suggests that the Cu2+ affinity site binds a histidine and to the of that data is for both His96 and His111 binding a Cu2+ His96 the Cu2+ as suggested by Prusiner S.B. J. 2003; PrP-(91–115) have the which a for coordination the and Furthermore, the affinity of Cu2+ for and PrP-(91–115) for Cu2+ are of than be for a and that the EPR Prusiner S.B. J. 2003; this in the complex of His111 a square-planar complex with different coordination In this are to a in In to the EPR of the Cu2+ binding has been used to J. J. Biol. 2001; Scholar). is a for a However, it is of square-planar coordination and in the between and is these can the of In the data physiological a which we have shown that than coordination is This of on these the that copper to both His111 and His96 J. J. Biol. 2001; Scholar). The of J. J. Biol. 2001; suggests and the of a the the of in Our NMR data indicates a of the to the on of The be to the copper coordination to the copper must be an range on the of EPR Indeed, to the coordination a role in and of Cu2+ J. Scholar). that His96 and His111 are in is R. von A. S. T. J. Biol. 1999; Scholar). The to a is by the Cu2+ it be a of is in the of as the have been spectroscopic studies on binding to the fragment M. R. 2001; Scholar, R. H. M. M. D. Scholar). However, the fragment have and is a good model for this unstructured region of the full-length PrP as Cu2+ to the Cu2+ coordination to the of in different used in this It is that copper coordination to PrP is to Cu2+ binding and affinity can be by in Cu2+ can bind to both the octarepeat region and PrP-(91–115) region in the the protein is the Cu2+ be the octarepeat region could be by the PrP-(91–115) region The of PrP-(91–115) Cu2+ binding and suggests that the region residues could a role in endocytosis of PrP. It has been shown that the octarepeat region is for endocytosis W.S. Hooper N.M. Curr. Biol. 2001; 11: 519-523Google Scholar). binding to the unstructured region outside the octarepeat a role in the of the full-length prion protein. this that the region between the octarepeat and the as a binding to could the octarepeat region for copper binding. Indeed, the octarepeat region to residues between 90 and the affinity and of Cu2+ binding to the A. S. S. Kretzschmar H. J. Biol. Chem. 2001; Scholar, Cohen F.E. A.L. Prusiner S.B. Sci. Scholar). Studies that show residues are for prion propagation T. M. Cohen F.E. Prusiner S.B. Proc. Natl. Acad. Sci. U. S. A. Scholar, M. T. A. A. M. S. A. J. Scholar, T. M. Cohen F.E. Prusiner S.B. 1997; the between disease and Cu2+ binding to the octarepeat region However, residues in have been shown to be vital for prion propagation T. M. Cohen F.E. Prusiner S.B. Proc. Natl. Acad. Sci. U. S. A. Scholar, M. T. A. A. M. S. A. J. Scholar, T. M. Cohen F.E. Prusiner S.B. 1997; Scholar). binding of Cu2+ to this region of PrP requires to the role of copper in prion misfolding and The of a high affinity Cu2+ site have on the structure in this region of PrPC, and can for the of cellular in vivo K. Kretzschmar Fraser P.E. Westaway D. J. Biol. Chem. Scholar). to in different strains of prion disease S. J. Biol. 1999; Scholar). the we have shown that coordination of a Cu2+ to both His96 and His111 has a on the of this region a β-sheet conformation. It is this region of PrP that is to be to prion misfolding and and data suggests a role for Cu2+ in this misfolding and for NMR and for the of NMR for with EPR and for the of the with
Jones et al. (Thu,) studied this question.