Structural insights into alternate aggregated prion protein forms.
The conversion of the cellular form of the prion protein (PrP(C)) to an abnormal, alternatively folded isoform (PrP(Sc)) is the central event in prion diseases or transmissible spongiform encephalopathies. Recent studies have demonstrated de novo generation of murine prions from recombinant prion protein (recPrP) after inoculation into transgenic and wild-type mice. These so-called synthetic prions lead to novel prion diseases with unique neuropathological and biochemical features. Moreover, the use of recPrP in an amyloid seeding assay can specifically detect and amplify various strains of prions. We employed this assay in our experiments and analyzed in detail the morphology of aggregate structures produced under defined chemical constraints. Our results suggest that changes in the concentration of guanidine hydrochloride can lead to different kinetic traces in a typical thioflavin T(ThT) assay. Morphological and structural analysis of these aggregates by atomic force microscopy indicates a variation in the structure of the PrP molecular assemblies. In particular, ThT positive PrP aggregates produced from rec mouse PrP residues 89 to 230 lead to mostly oligomeric structures at low concentrations of guanidine hydrochloride, while more amyloidal structures were observed at higher concentrations of the denaturant. These findings highlight the presence of numerous and complex pathways in deciphering prion constraints for infectivity and toxicity.
Polano
Maurizio
M
Laboratory of Prion Biology, Neurobiology Sector, Scuola Internazionale Superiore di Studi Avanzati-International School of Advanced Studies (SISSA-ISAS) Edificio Q1, Area Science Park, SS 14 Km 163.5, I-34149 Basovizza (TS), Italy.
Bek
Alpan
A
Benetti
Federico
F
Lazzarino
Marco
M
Legname
Giuseppe
G
D000682:Amyloid / Q000737:chemistry / Q000648:ultrastructure; D000818:Animals; D019791:Guanidine / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D018625:Microscopy, Atomic Force; D011328:Prions / Q000737:chemistry* / Q000648:ultrastructure; D011489:Protein Denaturation / Q000187:drug effects; D020033:Protein Isoforms / Q000737:chemistry / Q000648:ultrastructure; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry; D013499:Surface Properties / Q000187:drug effects
D000682:Amyloid; D011328:Prions; D020033:Protein Isoforms; D011994:Recombinant Proteins; D019791:Guanidine