Prion protein (PrP) synthetic peptides induce cellular PrP to acquire properties of the scrapie isoform.
Conversion of the cellular isoform of prion protein (PrPC) into the scrapie isoform (PrPSc) involves an increase in the beta-sheet content, diminished solubility, and resistance to proteolytic digestion. Transgenetic studies argue that PrPC and PrPSc form a complex during PrPSc formation; thus, synthetic PrP peptides, which mimic the conformational pluralism of PrP, were mixed with PrPC to determine whether its properties were altered. Peptides encompassing two alpha-helical domains of PrP when mixed with PrPC produced a complex that displayed many properties of PrPSc. The PrPC-peptide complex formed fibrous aggregates and up to 65% of complexed PrPC sedimented at 100,000 x g for 1 h, whereas PrPC alone did not. These complexes were resistant to proteolytic digestion and displayed a high beta-sheet content. Unexpectedly, the peptide in a beta-sheet conformation did not form the complex, whereas the random coil did. Addition of 2% Sarkosyl disrupted the complex and rendered PrPC sensitive to protease digestion. While the pathogenic A117V mutation increased the efficacy of complex formation, anti-PrP monoclonal antibody prevented interaction between PrPC and peptides. Our findings in concert with transgenetic investigations argue that PrPC interacts with PrPSc through a domain that contains the first two putative alpha-helices. Whether PrPC-peptide complexes possess prion infectivity as determined by bioassays remains to be established.
D000818:Animals; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D016466:CHO Cells; D006224:Cricetinae; D019286:Endopeptidase K; D008647:Mesocricetus; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry; D011328:Prions / Q000737:chemistry*; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D012608:Scrapie / Q000503:physiopathology*; D012697:Serine Endopeptidases / Q000378:metabolism; D012995:Solubility; D013045:Species Specificity; D017550:Spectroscopy, Fourier Transform Infrared
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D011328:Prions; D012697:Serine Endopeptidases; D019286:Endopeptidase K