Raman optical activity and circular dichroism reveal dramatic differences in the influence of divalent copper and manganese ions on prion protein folding

F Zhu, P Davies, A R Thompsett, S M Kelly, G E Tranter, L Hecht, N W Isaacs, D R Brown, L D Barron

Research output: Contribution to journalArticle

34 Citations (Scopus)

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 α-helix is lost, with UV CD revealing a drop from ∼25% to ∼18% in the total α-helix content. The lost α-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 α-helix content to ∼5%. Hence, although Cu2+ binding takes place exclusively within the unfolded/disordered N-terminal region, it can profoundly affect the structure of the folded/α-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 α-helix content to the metal-free protein. In contrast, when the protein is refolded in the presence of divalent manganese ions, ROA indicates the α-helix is reinforced, with UV CD revealing an increase in total α-helix content to ∼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.
Original languageEnglish
Pages (from-to)2510-2517
Number of pages8
JournalBiochemistry
Volume47
Issue number8
DOIs
Publication statusPublished - 26 Feb 2008

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