title: Raman optical activity and circular dichroism reveal dramatic differences in the influence of divalent copper and manganese ions on prion protein folding.
abstract: The binding of divalent copper ions to the full-length recombinant murine prion protein PrP23-231 at neutral pH was studied using vibrational Raman optical activity (ROA) and ultraviolet circular dichroism (UV CD). The effect of the Cu2+ ions on PrP structure depends on whether they are added after refolding of the protein in water or are present during the refolding process. In the first case ROA reveals that the hydrated alpha-helix is lost, with UV CD revealing a drop from approximately 25% to approximately 18% in the total alpha-helix content. The lost alpha-helix could be that comprising residues 145-156, located within the region associated with scrapie PrP formation. In the second case, ROA reveals the protein's structure to be almost completely disordered/irregular, with UV CD revealing a drop in total alpha-helix content to approximately 5%. Hence, although Cu2+ binding takes place exclusively within the unfolded/disordered N-terminal region, it can profoundly affect the structure of the folded/alpha-helical C-terminal region. This is supported by the finding that refolding in the presence of Cu2+ of a mutant in which the first six histidines associated with copper binding to the N-terminal region are replaced by alanine has a similar alpha-helix content to the metal-free protein. In contrast, when the protein is refolded in the presence of divalent manganese ions, ROA indicates the alpha-helix is reinforced, with UV CD revealing an increase in total alpha-helix content to approximately 30%. The very different influence of Cu2+ and Mn2+ ions on prion protein structure may originate in the different stability constants and geometries of their complexes.
authors.affiliation: WestChem, Department of Chemistry, University of Glasgow, Glasgow G12 8QQ, United Kingdom.
authors.forename: Fujiang
authors.initials: F
authors.lastname: Zhu
authors.forename: Paul
authors.initials: P
authors.lastname: Davies
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Thompsett
authors.forename: Sharon M
authors.initials: SM
authors.lastname: Kelly
authors.forename: George E
authors.initials: GE
authors.lastname: Tranter
authors.forename: Lutz
authors.initials: L
authors.lastname: Hecht
authors.forename: Neil W
authors.initials: NW
authors.lastname: Isaacs
authors.forename: David R
authors.initials: DR
authors.lastname: Brown
authors.forename: Laurence D
authors.initials: LD
authors.lastname: Barron
mesh_terms: D000818:Animals; D002413:Cations, Divalent / Q000378:metabolism / Q000494:pharmacology; D002942:Circular Dichroism; D003300:Copper / Q000378:metabolism / Q000494:pharmacology*; D020450:Heavy Ions; D046911:Macromolecular Substances / Q000737:chemistry; D008345:Manganese / Q000378:metabolism / Q000494:pharmacology*; D051379:Mice; D008958:Models, Molecular; D011328:Prions / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D017510:Protein Folding; D013059:Spectrum Analysis, Raman
chemical_list: D002413:Cations, Divalent; D046911:Macromolecular Substances; D011328:Prions; D008345:Manganese; D003300:Copper