title: Novel assay with fluorescence-labelled PrP peptides for differentiating L-type atypical and classical BSEs, and scrapie.
abstract: Characteristic differences of prions may account for the conformational diversity of the pathogenic isoform of prion protein (PrP(Sc)). Here, we applied a protein detection procedure by using fluorescent-labelled peptides for detecting PrP(Sc). Five prion protein (PrP) related peptides were found to change significantly their fluorescent intensities with prion-affected animal samples. Their reactivity was different among atypical L-BSE, classical BSE and scrapie. The pull-down assay revealed that they precipitated PrP(Sc) specifically. These findings suggest that fluorescent intensity changes depend on peptide-PrP(Sc) binding. This novel approach may distinguish the fine structural differences in PrP(Sc), which were not detected by the pull-down assay.
authors.affiliation: Prion Disease Research Center, National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Kasai
authors.forename: Akiyoshi
authors.initials: A
authors.lastname: Hirata
authors.forename: Takafumi
authors.initials: T
authors.lastname: Ohyama
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Nokihara
authors.forename: Takashi
authors.initials: T
authors.lastname: Yokoyama
authors.forename: Shirou
authors.initials: S
authors.lastname: Mohri
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D003937:Diagnosis, Differential; D016643:Encephalopathy, Bovine Spongiform / Q000145:classification / Q000175:diagnosis*; D005456:Fluorescent Dyes; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D018620:PrPSc Proteins / Q000737:chemistry / Q000235:genetics; D011328:Prions / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D012608:Scrapie / Q000175:diagnosis*; D012756:Sheep
chemical_list: D005456:Fluorescent Dyes; D010446:Peptide Fragments; D018620:PrPSc Proteins; D011328:Prions