Reconstruction of infectious prion protein in vitro

2010 Fiscal Year Final Research Report

• Project/Area Number: 21790846
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Neurology
• Research Institution: Nagasaki University
• Principal Investigator: SANO Kazunori Nagasaki University, 大学院・医歯薬学総合研究科, 助教 (50534343)
• Project Period (FY): 2009 – 2010
• Keywords: 神経分子病態学

Research Abstract

Recently, we developed an in vitro recombinant PrP (recPrP) conversion system with shaking (Quaking-Induced Conversion ; QUIC). We generated recPrP-amyloid in vitro by the tiny amounts of mouse brain homogenates with Chandler and 22L strains using this system. The shape of β-sheet spectrum by infrared spectroscopy in these recPrP-amyloids was similar to that of PrP^<Sc> isolated from each infectious brain, and unique to the strains. Additionally, the conformational stability analysis using denaturation by guanidine-hydrochloride demonstrated that the stability of recPrP-amyloid was similar to that of PrP^<Sc> in brains, indicating that prion strain specific conformations are seemed to be conserved. To determine infectivity of recPrP amyloids, recPrP amyloids was inoculated intracerebrally into wild-type mice. recPrP amyloids significantly accelerated the onset of disease. These results suggest that prion strain-specific conformations are transmitted to recPrP-amyloid, which is PrP^<Sc>-like infectious agents.

Research Products

• [Journal Article] Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion (2011)
  • Author(s): Ryuichiro Atarashi, Katsuya Satoh, Kazunori Sano, Takayuki Fuse, Naohiro Yamaguchi, Daisuke Ishibashi, Takehiro Matsubara, Takehiro Nakagaki, Hitoki Yamanaka, Susumu Shirabe, Masahito Yamada, Hidehiro Mizusawa, Tetsuyuki Kitamoto, Genevieve Klug, Amelia McGlade, Steven J Collins, Noriyuki Nishida
  • Journal Title: Nature medicine 17巻 Pages: 175-178
  • Peer Reviewed
• [Journal Article] Rapid End-Point Quantitation of Prion Seeding Activity with Sensitivity Comparable to Bioassays (2010)
  • Author(s): Jason M.Wilham, Christina D.Orru, Richard A.Bessen, Ryuichiro Atarashi, Kazunori Sano, Brent Race, Kimberly D.Meade-White, Lara M.Taubner, Andrew Timmes, Byron Caughey
  • Journal Title: PLoS pathogens 6巻 Pages: e1001217
  • Peer Reviewed
• [Presentation] Reconstruction of infectious prion protein in vitro
  • Author(s): 佐野和憲
  • Organizer: 第83回日本生化学会大会
• [Remarks] ホームページ等