title: PK-sensitive PrP is infectious and shares basic structural features with PK-resistant PrP.
abstract: One of the main characteristics of the transmissible isoform of the prion protein (PrP(Sc)) is its partial resistance to proteinase K (PK) digestion. Diagnosis of prion disease typically relies upon immunodetection of PK-digested PrP(Sc) following Western blot or ELISA. More recently, researchers determined that there is a sizeable fraction of PrP(Sc) that is sensitive to PK hydrolysis (sPrP(Sc)). Our group has previously reported a method to isolate this fraction by centrifugation and showed that it has protein misfolding cyclic amplification (PMCA) converting activity. We compared the infectivity of the sPrP(Sc) versus the PK-resistant (rPrP(Sc)) fractions of PrP(Sc) and analyzed the biochemical characteristics of these fractions under conditions of limited proteolysis. Our results show that sPrP(Sc) and rPrP(Sc) fractions have comparable degrees of infectivity and that although they contain different sized multimers, these multimers share similar structural properties. Furthermore, the PK-sensitive fractions of two hamster strains, 263K and Drowsy (Dy), showed strain-dependent differences in the ratios of the sPrP(Sc) to the rPrP(Sc) forms of PrP(Sc). Although the sPrP(Sc) and rPrP(Sc) fractions have different resistance to PK-digestion, and have previously been shown to sediment differently, and have a different distribution of multimers, they share a common structure and phenotype.
authors.affiliation: Department of Medicine, School of Medicine, University of Santiago de Compostela, Santiago de Compostela, Galiza, Spain. gsajnani@sescam.jccm.es
authors.forename: Gustavo
authors.initials: G
authors.lastname: Sajnani
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Silva
authors.forename: Adriana
authors.initials: A
authors.lastname: Ramos
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Pastrana
authors.forename: Bruce C
authors.initials: BC
authors.lastname: Onisko
authors.forename: Melissa L
authors.initials: ML
authors.lastname: Erickson
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Antaki
authors.forename: Irina
authors.initials: I
authors.lastname: Dynin
authors.forename: Ester
authors.initials: E
authors.lastname: Vázquez-Fernández
authors.forename: Christina J
authors.initials: CJ
authors.lastname: Sigurdson
authors.forename: J Mark
authors.initials: JM
authors.lastname: Carter
authors.forename: Jesús R
authors.initials: JR
authors.lastname: Requena
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology; D006224:Cricetinae; D004195:Disease Models, Animal; D019286:Endopeptidase K / Q000378:metabolism*; D008136:Longevity; D008647:Mesocricetus; D018620:PrPSc Proteins / Q000378:metabolism*; D011487:Protein Conformation; D011815:R Factors; D012608:Scrapie / Q000201:enzymology*
chemical_list: D018620:PrPSc Proteins; D019286:Endopeptidase K