Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins.
Prions are composed largely, if not entirely, of prion protein (PrPSc in the case of scrapie). Although the formation of PrPSc from the cellular prion protein (PrPC) is a post-translational process, no candidate chemical modification was identified, suggesting that a conformational change features in PrPSc synthesis. To assess this possibility, we purified both PrPC and PrPSc by using nondenaturing procedures and determined the secondary structure of each. Fourier-transform infrared (FTIR) spectroscopy demonstrated that PrPC has a high alpha-helix content (42%) and no beta-sheet (3%), findings that were confirmed by circular dichroism measurements. In contrast, the beta-sheet content of PrPSc was 43% and the alpha-helix 30% as measured by FTIR. As determined in earlier studies, N-terminally truncated PrPSc derived by limited proteolysis, designated PrP 27-30, has an even higher beta-sheet content (54%) and a lower alpha-helix content (21%). Neither PrPC nor PrPSc formed aggregates detectable by electron microscopy, while PrP 27-30 polymerized into rod-shaped amyloids. While the foregoing findings argue that the conversion of alpha-helices into beta-sheets underlies the formation of PrPSc, we cannot eliminate the possibility that an undetected chemical modification of a small fraction of PrPSc initiates this process. Since PrPSc seems to be the only component of the "infectious" prion particle, it is likely that this conformational transition is a fundamental event in the propagation of prions.
Pan
K M
KM
Department of Neurology, University of California, San Francisco 94143.
Baldwin
M
M
Nguyen
J
J
Gasset
M
M
Serban
A
A
Groth
D
D
Mehlhorn
I
I
Huang
Z
Z
Fletterick
R J
RJ
Cohen
F E
FE
D000818:Animals; D002942:Circular Dichroism; D006224:Cricetinae; D007150:Immunohistochemistry; D008647:Mesocricetus; D008854:Microscopy, Electron; D018620:PrPSc Proteins; D011328:Prions / Q000737:chemistry* / Q000302:isolation & purification; D017433:Protein Structure, Secondary; D012608:Scrapie / Q000209:etiology*; D017550:Spectroscopy, Fourier Transform Infrared
D018620:PrPSc Proteins; D011328:Prions