title: Manganese binding to the prion protein.
abstract: There is considerable evidence that the prion protein binds copper. However, there have also been suggestions that prion protein (PrP) binds manganese. We used isothermal titration calorimetry to identify the manganese binding sites in wild-type mouse PrP. The protein showed two manganese binding sites with affinities that would bind manganese at concentrations of 63 and 200 mum at pH 5.5. This indicates that PrP binds manganese with affinity similar to other known manganese-binding proteins. Further study indicated that the main manganese binding site is associated with His-95 in the so-called "fifth site" normally associated with copper binding. Additionally, it was shown that occupancy by copper does not prevent manganese binding. Under these conditions, manganese binding resulted in an altered conformation of PrP, displacement of copper, and altered redox chemistry of the metal-protein complex. Cyclic voltammetric measurements suggested a complex redox chemistry involving manganese bound to PrP, whereas copper-bound PrP was able to undergo fully reversible electron cycling. Additionally, manganese binding to PrP converted it to a form able to catalyze aggregation of metal-free PrP. These results further support the notion that manganese binding could cause a conformation change in PrP and trigger changes in the protein similar to those associated with prion disease.
authors.affiliation: Department of Biology and Biochemistry and Chemistry, University of Bath, Bath BA2 7AY, UK.
authors.forename: Marcus W
authors.initials: MW
authors.lastname: Brazier
authors.forename: Paul
authors.initials: P
authors.lastname: Davies
authors.forename: Esmie
authors.initials: E
authors.lastname: Player
authors.forename: Frank
authors.initials: F
authors.lastname: Marken
authors.forename: John H
authors.initials: JH
authors.lastname: Viles
authors.forename: David R
authors.initials: DR
authors.lastname: Brown
mesh_terms: D000818:Animals; D001665:Binding Sites; D002151:Calorimetry; D002942:Circular Dichroism; D003300:Copper / Q000737:chemistry / Q000378:metabolism; D004563:Electrochemistry; D006863:Hydrogen-Ion Concentration; D008345:Manganese / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D009154:Mutation / Q000235:genetics; D011328:Prions / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D011328:Prions; D008345:Manganese; D003300:Copper