Hydrogen peroxide cleavage of the prion protein generates a fragment able to initiate polymerisation of full length prion protein.
The prion protein is central to the disease pathogenesis of a variety of neurodegenerative diseases such as CJD. The protein is only able to initiate the disease process following post-translational modification. The main characteristic of this change is the ability of this altered isoform to polymerise. We wish to determine if altered cleavage of the protein could generate a protein fragment able to initiate polymerisation. During normal metabolic breakdown the protein is initially cleaved at a single site at around amino acid residue 111/112 in the mouse sequence. A second site before amino acid residue 90 has been postulated as an alternative cleavage point. We have provided evidence that hydrogen peroxide as low as 50 microM in the presence of copper, iron or manganese (but not nickel, magnesium or zinc) can cleave the recombinant protein near this site and requires a GXXH motif in the protein sequence. This reaction results in the production of 6 and 19 kDa fragments of the protein. This cleavage pattern occurs in prion proteins from different species (mouse, chicken and turtle) and is enhanced by modification of the octameric repeat region. The 19 kDa fragment produced by this reaction is protease sensitive. This fragment in a pure form caused the polymerisation of wild-type prion protein by a seeding mechanism. Therefore our results provide a possible mechanism by which altered cleavage of the prion protein could result in the kind of protein polymerisation associated with prion diseases.
Abdelraheim
Salama R
SR
Department of Biology and Biochemistry, University of Bath, Calverton Down, Bath BA2 7AY, United Kingdom.
Královicová
Silvia
S
Brown
David R
DR
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002645:Chickens; D003300:Copper / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis / Q000187:drug effects; D015151:Immunoblotting; D007501:Iron / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*; D011328:Prions / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013696:Temperature; D014426:Turtles
D050505:Mutant Proteins; D010446:Peptide Fragments; D011108:Polymers; D011328:Prions; D011994:Recombinant Proteins; D008345:Manganese; D003300:Copper; D006861:Hydrogen Peroxide; D007501:Iron